Abomasal Infusion Research Articles

Abomasal or ruminal administration of flax oil and hulls on milk production, digestibility, and milk fatty acid profile of dairy cows

Four ruminally fistulated primiparous lactating Holstein cows were assigned to a 4 × 4 Latin square design to determine the effects of the site of administration (rumen or abomasum) of flax oil and flax hulls on diet digestibility and milk fatty acid profile of dairy cows. The treatments were 1) oil and hulls administered in the rumen and abomasal infusion of water (RUM/RUM), 2) oil and hulls infused in the abomasum (ABO/ABO), 3) oil administered in the rumen and hulls infused in the abomasum (RUM/ABO), and 4) oil infused in the abomasum and hulls placed in the rumen (ABO/RUM). Cows on the ABO/ABO and RUM/RUM treatments had the highest and lowest amounts of basal dry matter eaten, respectively. Higher dry matter digestibility was obtained when flax oil bypassed the rumen (ABO/ABO and ABO/RUM) compared with when flax oil was administered directly in the rumen (RUM/ABO and RUM/RUM). Apparent digestibility of ether extract was higher when flax hulls were administered in the rumen (RUM/RUM and ABO/RUM) compared with when flax hulls were infused in the abomasum (ABO/ABO and RUM/ABO). The lowest digestibility of acid detergent fiber and neutral detergent fiber was obtained when both flax products were added in the rumen, which may be attributed to the high amount of oil present in the rumen (7.8% of total dry matter input). The lowest yield of 4% fat-corrected milk was obtained for cows on the RUM/RUM treatment, probably as a result of lower dry matter intake and digestibility of fiber. Milk concentrations of protein, fat, total solids, and lactose were similar among treatments. Administration of oil and hulls in the rumen resulted in the highest concentrations of intermediate products of biohydrogenation and total trans fatty acids in milk fat compared with the other treatments. All ratios of n-6 to n-3 fatty acids in milk fat were lower than the 4 to 1 ratio recommended to improve human health. These results suggest that the presence of both flax oil and flax hulls in the rumen decreases 4% fat-corrected milk yield and digestibility but provides a desirable fatty acid profile of milk to enhance consumers’ health.

Regulation of protein synthesis in mammary glands of lactating dairy cows by starch and amino acids

The objective of this study was to evaluate local molecular adaptations proposed to regulate protein synthesis in the mammary glands. It was hypothesized that AA and energy-yielding substrates independently regulate AA metabolism and protein synthesis in mammary glands by a combination of systemic and local mechanisms. Six primiparous mid-lactation Holstein cows with ruminal cannulas were randomly assigned to 4 treatment sequences in a replicated incomplete 4 × 4 Latin square design experiment. Treatments were abomasal infusions of casein and starch in a 2 × 2 factorial arrangement. All animals received the same basal diet (17.6% crude protein and 6.61 MJ of net energy for lactation/kg of DM) throughout the study. Cows were restricted to 70% of ad libitum intake and abomasally infused for 36h with water, casein (0.86 kg/d), starch (2 kg/d), or a combination (2 kg/d starch+0.86 kg/d casein) using peristaltic pumps. Milk yields and composition were assessed throughout the study. Arterial and venous plasma samples were collected every 20min during the last 8h of infusion to assess mammary uptake. Mammary biopsy samples were collected at the end of each infusion and assessed for the phosphorylation state of selected intracellular signaling molecules that regulate protein synthesis. Animals infused with casein had increased arterial concentrations of AA, increased mammary extraction of AA from plasma, either no change or a trend for reduced mammary AA clearance rates, and no change in milk protein yield. Animals infused with starch had increased milk and milk protein yields, increased mammary plasma flow, reduced arterial concentrations of AA, and increased mammary clearance rates and net uptake of some AA. Infusions of starch increased plasma concentrations of glucose, insulin, and insulin-like growth factor-I. Starch infusions increased phosphorylation of ribosomal protein S6 and endothelial nitric oxide synthase, consistent with changes in milk protein yields and plasma flow, respectively. Phosphorylation of the mammalian target of rapamycin was increased in response to starch only when casein was also infused. Thus, cell signaling molecules involved in the regulation of protein synthesis differentially responded to these nutritional stimuli. The hypothesized independent effects of casein and starch on animal metabolism and cell signaling were not observed, presumably because of the lack of a milk protein response to infused casein.

Changes in production and mammary metabolism of dairy cows in response to essential and nonessential amino acid infusions

This study was undertaken to increase our understanding of the need of the mammary gland for the different types of AA and how the mammary gland alters its metabolism in response to a variable AA supply. Eight lactating Holstein cows (61±4 DIM) were used in a replicated 4 × 4 Latin square balanced for residual effects with 14-d periods. The diet was formulated to supply 100% of the net energy requirement and 72% of the metabolizable protein requirement. The 4 treatments were 1) abomasal infusions of water, 2) essential AA at 359g/d, 3) nonessential AA at 356g/d, and 4) essential AA at 359g/d+nonessential AA at 356g/d (total of 715g/d). The infusates had the same AA profile as casein with the exception that Met was increased to maintain a 3:1 ratio of digestible Lys to Met and because of solubility limitations all the Tyr was replaced by Phe and part of the Glu was replaced by Gln. Milk yield and milk protein yield were increased by the essential AA treatments compared with the other treatments. Mammary uptake of β-hydroxybutyrate plus lactate tended to increase with the essential AA treatments, whereas glucose mammary uptake tended to be higher with the nonessential AA treatments. With the essential AA treatments, the mammary uptake:milk protein output ratio for the group 1 AA (His, Met, Phe, Trp, and Tyr) did not differ from 1 but tended to increase; the ratio for the group 2 AA (Ile, Leu, Lys, and Val) did increase, significantly exceeding unity when essential AA were infused. These results indicate that the mammary gland alters differently its uptake of both AA and energy-yielding nutrients in response to the amount and profile of AA presented to it and that even under situations of protein deficiency nonessential AA supplementation does not enhance milk and milk protein synthesis.

The Effects of Graded Abomasal Glucose Infusion on Milk Yield and Composition in Lactating Goats

Four non-pregnant multiparous lactating goats were used in a 4 × 4 latin square experiment in the present study. The four treatments were abomasal infusion of 0, 50, 100, and 150 g d −1 glucose. The results showed that graded glucose infusion increased milk yield ( P < 0.01). The highest milk yield was observed when 50 g d −1 glucose was infused. An infusion of glucose at 100 or 150 g d −1 failed to increase milk yield further and milk yield of the 100 g d −1 glucose infusion treatment was even significantly lower than that of the 50 g d −1 treatment ( P < 0.01). Though milk fat content was gradually decreased by graded glucose infusions and milk fat yield of the 100 and 150 g d −1 treatments were lower than that of the control ( P < 0.01); milk fat yield of the 50 g d −1 treatment was not depressed and was higher than that of the control ( P < 0.01). Graded glucose infusion did not change the contents of milk protein and non-fatty solids. Yields of milk protein ( P < 0.05), lactose ( P < 0.01), non-fat solids ( P < 0.01), and lactose content ( P < 0.01) of the 50 g d −1 treatment were higher than those of the control but had no difference among the other three treatments. Contents of those de novo synthesized fatty acids in milk fat remained unchanged or increased and those preformed fatty acids remained unchanged or decreased by graded glucose infusion. Glucose infusion decreased the total yields of C16 + C16:1 and > C16 fatty acids and had no influence on the yield of C4-C14:1 fatty acids ( P < 0.05). Results of the present study implied that increased glucose supply had positive effects on the production of milk, milk protein, milk fat, and lactose only when glucose was a limiting factor for milk synthesis. Excessive glucose supply had no further beneficial effects and might depress milk fat synthesis.

In vitro and in vivo analysis of fatty acid effects on metabolism of 17β-estradiol and progesterone in dairy cows

Some studies have reported improved reproductive performance with dietary fat supplementation. This study examined effects of fatty acids with different lengths, or desaturation, or both, on metabolism of estradiol (E2) and progesterone (P4) in bovine liver slice incubations (experiments 1 and 2) and in vivo (experiment 3). In experiment 1, effects of fatty acids C16:0 (palmitic acid), C16:1 (palmitoleic acid), C18:1 (oleic acid), and C18:3 (linolenic acid) were evaluated at 30, 100, and 300 μM on P4 and E2 metabolism in vitro. In experiment 2, stearic acid (C18:0) and C18:3 were evaluated in the same incubation conditions. In experiment 1, all of the fatty acids had some significant inhibitory effect on metabolism of P4, E2, or both (300 μM C16:0 on E2; 100 μM C16:1 on E2; 300 μM C16:1 on both P4 and E2; 300 μM C18:1 on P4; and 100 and 300 μM C18:3 on both P4 and E2). In experiment 2, C18:3 (100 and 300 μM) but not C18:0 decreased P4 and E2 metabolism. Overall, the most profound increase (∼60%) in half-life of P4 and E2 was observed with incubations of 300 μM C18:3 in both in vitro experiments. Based on these in vitro results, in experiment 3 linseed oil (rich in C18:3) was supplemented into the abomasum and acute effects on metabolism of E2 and P4 were evaluated. Cows (n=4) had endogenous E2 and P4 minimized (corpus luteum regressed, follicles aspirated) before receiving continuous intravenous infusion of E2 and P4 to analyze metabolic clearance rate for these hormones during abomasal infusion of saline (control) or 70mL of linseed oil every 4h for 28h. Linseed oil infusion increased C18:3 in plasma by 46%; however, metabolic clearance rate for E2 and P4 were similar for control cows compared with linseed-treated cows. Thus, in vitro experiments indicated that E2 and P4 metabolism can be inhibited by high concentrations of C18:3. Nevertheless, in vivo, linseed oil did not acutely inhibit E2 and P4 metabolism, perhaps because insufficient C18:3 concentrations (increased to ∼8 μM) were achieved. Further research is needed to determine the mechanism(s) of fatty acid inhibition of P4 and E2 metabolism and to discover practical methods to mimic this effect in vivo.

Effects of glucose and amino acids on ghrelin secretion in sheep

Two experiments were conducted to elucidate the effects of post-ruminal administration of starch and casein (Exp. 1), plasma amino acids concentrations (Exp. 2), and plasma glucose and insulin concentrations (Exp. 2) on plasma ghrelin concentrations in sheep. In Exp. 1, plasma ghrelin concentrations were determined by four infusion treatments (water, cornstarch, casein and cornstarch plus casein) in four wethers. Abomasal infusion of casein increased plasma alpha-amino N (AAN) concentrations. Infusion of starch or casein alone did not affect plasma ghrelin concentrations, but starch plus casein infusion increased plasma levels of ghrelin, glucose and AAN. In Exp 2, we investigated the effects of saline or amino acids on ghrelin secretion in four wethers. Two hours after the initiation of saline or amino acid infusion into the jugular vein, glucose was also continuously infused to investigate the effects of blood glucose and insulin by hyper-glycemic clump on plasma ghrelin concentrations. Infusion of amino acids alone raised plasma levels of ghrelin, but the higher plasma glucose and insulin concentrations had no effect on plasma ghrelin concentrations. These results suggest that high plasma levels of amino acids can stimulate ghrelin secretion, but glucose and insulin do not affect ghrelin secretion in sheep.

Plasma ghrelin and oxyntomodulin concentrations in lactating dairy cows receiving abomasal soybean oil, corn starch, and casein infusions

The effects of increased postruminal supply of casein, corn starch, and soybean oil on plasma concentrations of the gastrointestinal hormones ghrelin and oxyntomodulin (OXM) were investigated. Four mid-lactation Holstein cows were used in a 4 × 4 Latin square. Treatments were continuous abomasal infusions (23 h/d) for 7 d of water, soybean oil (500 g/d), corn starch (1100 g/d), or casein (800 g/d). Jugular vein plasma was obtained every 30 min for 7 h on days 1 and 7. Soybean oil and casein infusion decreased preprandial plasma ghrelin concentration by approximately 20% on both d (time-by-treatment P < 0.10); however, dry matter intake (DMI) was depressed only after 7 d of oil infusion. Infusion of soybean oil, corn starch, or casein did not change the plasma OXM concentration ( P > 0.20 ). The present data indicate that plasma ghrelin concentration is depressed immediately before feeding by the postruminal infusion of soybean oil and casein, but it is not affected during the postprandial period. Plasma ghrelin concentration was not altered ( P > 0.20), pre- or postfeeding, by increased postruminal supply of corn starch. In addition, plasma OXM concentration did not respond ( P > 0.20 ) to postruminal nutrient infusion. In conclusion, a decrease in DMI when fat is infused could be partially explained by the decrease in prefeeding plasma ghrelin concentration, but a decrease in prefeeding plasma ghrelin concentration is not always associated with a decrease in DMI, as observed for the infusion of casein. Plasma OXM concentration was not affected by postruminal infusion of macronutrients.

Role of trans fatty acids in the nutritional regulation of mammary lipogenesis in ruminants

Fat is an important constituent contributing to the organoleptic, processing and physical properties of ruminant milk. Understanding the regulation of milk fat synthesis is central to the development of nutritional strategies to enhance the nutritional value of milk, decrease milk energy secretion and improve the energy balance of lactating ruminants. Nutrition is the major environmental factor regulating the concentration and composition of fat in ruminant milk. Feeding low-fibre/high-starch diets and/or lipid supplements rich in polyunsaturated fatty acids induce milk fat depression (MFD) in the bovine, typically increase milk fat secretion in the caprine, whereas limited data in sheep suggest that the responses are more similar to the goat than the cow. Following the observation that reductions in milk fat synthesis during diet-induced MFD are associated with increases in the concentration of specific trans fatty acids in milk, the biohydrogenation theory of MFD was proposed, which attributes the causal mechanism to altered ruminal lipid metabolism leading to increased formation of specific biohydrogenation intermediates that exert anti-lipogenic effects. Trans-10, cis-12 conjugated linoleic acid (CLA) is the only biohydrogenation intermediate to have been infused at the abomasum over a range of experimental doses (1.25 to 14.0 g/day) and shown unequivocally to inhibit milk fat synthesis in ruminants. However, increases in ruminal trans-10, cis-12 CLA formation do not explain entirely diet-induced MFD, suggesting that other biohydrogenation intermediates and/or other mechanisms may also be involved. Experiments involving abomasal infusions (g/day) in lactating cows have provided evidence that cis-10, trans-12 CLA (1.2), trans-9, cis-11 CLA (5.0) and trans-10 18:1 (92.1) may also exert anti-lipogenic effects. Use of molecular-based approaches have demonstrated that mammary abundance of transcripts encoding for key lipogenic genes are reduced during MFD in the bovine, changes that are accompanied by decrease in sterol response element binding protein 1 (SREBP1) and alterations in the expression of genes related to the SREBP1 pathway. Recent studies indicate that transcription of one or more adipogenic genes is increased in subcutaneous adipose tissue in cows during acute or chronic MFD. Feeding diets of similar composition do not induce MFD or substantially alter mammary lipogenic gene expression in the goat. The available data suggests that variation in mammary fatty acid secretion and lipogenic responses to changes in diet composition between ruminants reflect inherent interspecies differences in ruminal lipid metabolism and mammary specific regulation of cellular processes and key lipogenic enzymes involved in the synthesis of milk fat triacylglycerides.

Hot topic: Enhancing omega-3 fatty acids in milk fat of dairy cows by using stearidonic acid-enriched soybean oil from genetically modified soybeans

Very long chain n-3 fatty acids such as eicosapentaenoic acid (EPA; 20:5n-3) are important in human cardiac health and the prevention of chronic diseases, but food sources are limited. Stearidonic acid (SDA; 18:4n-3) is an n-3 fatty acid that humans are able to convert to EPA. In utilizing SDA-enhanced soybean oil (SBO) derived from genetically modified soybeans, our objectives were to examine the potential to increase the n-3 fatty acid content of milk fat and to determine the efficiency of SDA uptake from the digestive tract and transfer to milk fat. Three multiparous, rumen-fistulated Holstein cows were assigned randomly in a 3 × 3 Latin square design to the following treatments: 1) control (no oil infusion); 2) abomasal infusion of SDA-enhanced SBO (SDA-abo); and 3) ruminal infusion of SDA-enhanced SBO (SDA-rum). The SDA-enhanced SBO contained 27.1% SDA, 10.4% α-linolenic acid, and 7.2% γ-linolenic acid. Oil infusions provided 57 g/d of SDA with equal amounts of oil infused into either the rumen or abomasum at 6-h intervals over a 7-d infusion period. Cow numbers were limited and no treatment differences were detected for DMI or milk production (22.9±0.5 kg/d and 32.3±0.9 kg/d, respectively; least squares means ± SE), milk protein percentage and yield (3.24±0.04% and 1.03±0.02 kg/d), or lactose percentage and yield (4.88±0.05% and 1.55±0.05 kg/d). Treatment also had no effect on milk fat yield (1.36±0.03 kg/d), but milk fat percentage was lower for the SDA-rum treatment (4.04±0.04% vs. 4.30±0.04% for control and 4.41±0.05% for SDA-abo). The SDA-abo treatment increased n-3 fatty acids to 3.9% of total milk fatty acids, a value more than 5-fold greater than that for the control. Expressed as a percentage of total milk fatty acids, values (least squares means ± SE) for the SDA-abo treatment were 1.55±0.03% for α-linolenic acid (18:3n-3), 1.86±0.02 for SDA, 0.23 ± <0.01 for eicosatetraenoic acid (20:4n-3), and 0.18±0.01 for EPA. Transfer efficiency of SDA to milk fat represented 39.3% (range=36.8 to 41.9%) of the abomasally infused SDA and 47.3% (range=45.0 to 49.6%) when the n-3 fatty acids downstream from SDA were included. In contrast, transfer of ruminally infused SDA to milk fat averaged only 1.7% (range=1.3 to 2.1%), indicating extensive rumen biohydrogenation. Overall, results demonstrate the potential to use SDA-enhanced SBO from genetically modified soybeans combined with proper ruminal protection to achieve impressive increases in the milk fat content of SDA and other n-3 fatty acids that are beneficial for human health.

Net flux of amino acids across the portal-drained viscera and liver of the ewe during abomasal infusion of protein and glucose12

The objective of the study was to measure net AA flux rates across the portal-drained viscera (PDV) and liver in the presence and absence of abomasal glucose infusion. Decreasing the fraction of AA metabolized by the mucosal cells may increase the fraction of AA being released into the blood. A potential mechanism to reduce AA catabolism by mucosal cells is to provide an alternative source of energy. We hypothesized that increasing glucose flow to the small intestine would increase net appearance of AA across the PDV. Eighteen mature ewes with sampling catheters were placed on study. The experimental design was a split-plot with a complete randomized design on the whole-plot and a Latin-square subplot with 5 periods and incremental levels of protein infusion. One-half of the ewes received abomasal glucose infusions (3.84 g/h), and all ewes received each of 5 protein abomasal infusion levels over 5 periods (0, 2.6, 5.2, 7.8, and 10.5 g/h). Net PDV release of isoleucine, leucine, methionine, phenylalanine, aspartate, glutamate, glutamine, proline, serine, and tyrosine increased linearly with increased protein infusion, and net PDV release of histidine, lysine, threonine, valine, alanine, and glycine did not differ with protein infusion. Net hepatic glucose release decreased with glucose infusion. With the exception of histidine, phenylalanine, and valine, net hepatic AA uptake increased linearly with increased delivery of AA to the liver. Glucose infusion increased the hepatic lysine and valine uptake and decreased phenylalanine uptake. Based on the observations in the current study, we reject our hypothesis that glucose can spare AA metabolism by PDV tissue. Our findings suggest that hepatic gluconeogenesis can be increased in the presence of increased AA delivery to the liver and that hepatic gluconeogenesis can be decreased with increased absorption of dietary glucose. Our findings support the concept that for most AA, hepatic transport of AA can be described by mass action kinetics; however, the rates of hepatic uptake of specific AA are upregulated directly or indirectly by elevated glucose.

Effects of abomasal lipid infusion on liver triglyceride accumulation and adipose lipolysis during fatty liver induction in dairy cows

The objective was to determine the effects of abomasal infusion of linseed oil on liver triglyceride (TG) accumulation and adipose tissue lipolysis during an experimental protocol for induction of fatty liver. Eight nonpregnant, nonlactating Holstein cows were randomly assigned to treatments in a replicated 4×4 Latin square design. Treatments were abomasal infusion of water (W), tallow (T), linseed oil (LO), or half linseed oil and half tallow (LOT) at a rate of 0.56g/kg of body weight per day. Each experimental period consisted of a 4-d fast concurrent with administration of treatments into the abomasum in 6 equal doses per day (every 4h). Cows were fed ad libitum for 24 d between periods of fasting and lipid infusion. Infusion of linseed oil (LO and LOT) increased α-linolenic acid (C18:3n-3) content in serum (12.2, 10.4, 4.2, and 4.6g/100g of fatty acids for LO, LOT, T, and W, respectively), but not in the nonesterified fatty acid (NEFA) fraction of plasma. Treatments had no effect on plasma NEFA concentrations. Abomasal infusion of lipid increased in vitro stimulated lipolysis in subcutaneous adipose tissue, compared with W (4,294, 3,809, 4,231, and 3,293 nmol of glycerol released×g−1 tissue×2h−1 for LO, LOT, T, and W, respectively), but there was no difference between fat sources. Hepatic TG accumulation over 4-d fast was 2.52, 2.60, 2.64, and 2.09±0.75μg of TG/μg of DNA for W, LO, LOT, and T, respectively, which did not differ. Abomasal infusion of LO did not reduce liver TG accumulation, plasma NEFA concentration, or alter in vitro adipose tissue lipolysis when compared with T. These results contrast with a previous study involving i.v. infusion of lipid emulsion derived from LO. Discrepancies might be explained by the use of different administration routes and a relatively modest induction of liver TG accumulation in the current experiment.

Milk production responses of primiparous and multiparous dairy cows to dose of conjugated linoleic acid consumed in rumen inert form

Conjugated linoleic acid (CLA) has the potential to be used as a dietary means of manipulating milk fat production of dairy cows to meet specified short-term targets, as in a supply-managed industry. An experiment was conducted to evaluate the dose-response to calcium salts of CLA fed as a top-dress on a corn- and alfalfa-based TMR. In two 5 × 5 Latin squares of 28-d periods, five primiparous and five multiparous cows were fed 0, 22, 45, 67 or 178 g d-1 CLA (29% trans-10, cis-12) as a calcium salt. Results from one multiparous cow on the 178 g d-1 treatment were removed from statistical analysis because milk production fell to 1.2 kg d-1 by the end of the period. On average, milk, lactose and protein yields increased at the lower doses of CLA and decreased to control levels at the highest dose. Milk fat yield declined progressively from 876 to 770, 689, 676 and 543 g d-1 on the five doses. Net energy balance of cows increased linearly from -1.5 Mcal d-1 on the control to 2.0 Mcal d-1 at 178 g d-1 CLA. The percentage depression in milk fat yield at different doses of trans-10, cis-12 CLA was described by the equation y = 54/(1 + 22.8/dose). Comparison with published literature suggested that feeding calcium salts of CLA was 30% as effective in depressing milk fat production as an abomasal infusion, due to rumen biohydrogenation and intestinal indigestibility.Key words: Milk composition, cattle, diet

Effect of abomasal infusions of a mixture of octadecenoic acids on milk fat synthesis in lactating cows

Diets causing milk fat depression (MFD) are known to alter ruminal lipid metabolism leading to the formation of specific biohydrogenation intermediates that exert antilipogenic effects. Several isomers of conjugated linoleic acid (CLA), namely trans-10, cis-12 CLA, cis-10, trans-12 CLA, and trans-9, cis-11 CLA, inhibit mammary lipogenesis in the lactating cow, but ruminal outflow of these biohydrogenation intermediates does not account entirely for the reductions in milk fat synthesis during diet-induced MFD. Milk fat trans-10 18:1 concentrations are consistently increased on diets that cause MFD, suggesting a possible role in the regulation of milk fat secretion. Three rumen-fistulated cows in mid lactation were used in a 3×3 Latin square to evaluate the effects of a mixture of 18:1 fatty acid methyl esters (FAME) on milk fat synthesis. Experimental treatments consisted of abomasal infusions of ethanol (control), 6 g/d of trans-10, cis-12 CLA (positive control; CLA), or 247 g/d of a mixture of 18:1 FAME containing (% fatty acids) cis-9 (9.45), cis-12 (3.35), trans-10 (37.3), trans-11 (37.4), and trans-12 (2.66) as major isomers (T181 treatment). Administration of the T181 treatment supplied 92.1 g/d of trans-10 18:1. Infusions were conducted over a 5-d period with a 9-d interval between treatments. Treatments had no effect on dry matter intake, milk yield, or milk protein. Relative to the control, abomasal infusion of T181 and trans-10, cis-12 CLA treatments reduced milk fat secretion by 19.5 and 41.5%, respectively. Even though a direct cause and effect on mammary lipogenesis could not be established, comparisons with published data and considerations of the relative abundance of constituent FAME in treatment T181 implicated trans-10 18:1 as the isomer responsible. In conclusion, current data suggest that trans-10 18:1 potentially exerts antilipogenic effects and may contribute to the reduction in milk fat synthesis during diet-induced MFD in the lactating cow.

Effects of abomasal pectin infusion on milk production, digestion and nitrogen utilization pattern of lactating Saanen dairy goats

To study the effects of graded abomasal infusion of pectin on milk production, nitrogen balance and nutrient digestibilities of goats, four abomasally cannulated lactating Sannen dairy goats were used in a 4 × 4 latin square design with 14-d periods. Goats were fed the same basal diet and the treatments were the abomasal infusion of 0, 40, 80 or 120 g/d of citrus pectin. Pectin infusion resulted in linear decreases in basal ration intake from 2250 to 1985 g/d. There was insignificant decrease in total tract apparent digestibilities of dry matter and neutral detergent fiber. Decreased basal ration intake and digestibility led to decreased milk production from 1.95 to 1.75 kg/d. Milk fat content increased quadratically with increasing levels of infused pectin ( P = 0.04), but milk fat yield was unaffected by treatments. Milk total solids and solid-not-fat were also linearly decreased by pectin infusion. There were linear decreases in urinary N and plasma urea N (PUN) with the increase in infusion of pectin. As a proportion of N intake, urinary N excretion decreased from 36.31 to 31.92%, whereas N excreted from faeces increased from 25.2 to 29.4%, as the amount of pectin infusion increased. Abomasal pectin linearly decreased faecal pH from 7.31 to 6.86 and tended to decrease faecal ammonia from 0.246 to 0.215 mg/g faecal DM. These results suggest that manipulating dairy goats diets to increase postruminal fermentation may reduce urinary N and consequently manure ammonia losses. However, decreased digestibility and milk production at the highest level of pectin infusion is suggesting that pectin may have reduced postruminal nutrient utilization.

Expression of Enzymes and Key Regulators of Lipid Synthesis Is Upregulated in Adipose Tissue during CLA-Induced Milk Fat Depression in Dairy Cows1–3

Milk fat depression (MFD) is a naturally occurring condition in dairy cows where milk fat synthesis is inhibited by intermediates of ruminal biohydrogenation. One of these bioactive fatty acids (FA), trans-10, cis-12 conjugated linoleic acid (CLA), decreases milk fat synthesis through transcriptional downregulation of genes involved in mammary lipid synthesis. Energy partitioning during MFD is not well characterized because of the complexity of observing energy metabolism in ruminant animals. To investigate energy partitioning during MFD, adipose tissue biopsies were taken from 4 cows arranged in a switchback design. Treatments were control and 4-d abomasal infusion of trans-10, cis-12 CLA (7.5 g/d). CLA decreased milk fat yield by 38% and milk fat content by 34%, but yields of milk and other milk components were unchanged. In contrast to reported changes in mammary tissue, adipose tissue expression of lipid synthesis enzymes, including lipoprotein lipase, FA synthase, stearoyl-CoA desaturase, and FA binding protein 4, was increased. Expression of regulators of lipid synthesis, including sterol-response element binding protein 1, thyroid hormone responsive spot 14, and PPARγ, also increased in adipose tissue. Thus, a CLA dose resulting in near maximal inhibition of mammary lipid synthesis resulted in increased expression of lipid synthesis-related genes in adipose tissue. A meta-analysis of intake response during CLA infusion was conducted to extend the investigation of energy metabolism during MFD. Voluntary intake decreased (P < 0.001) by 1.5 kg/d during CLA-induced MFD in the 14 studies analyzed, but the reduction in intake only partially accounts for the energy spared from reduced milk fat synthesis. Results are consistent with energy spared from the reduction in milk fat synthesis being partitioned toward adipose tissue fat stores during short-term MFD.

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.

Ruminal and abomasal starch hydrolysate infusions selectively decrease the expression of cationic amino acid transporter mRNA by small intestinal epithelia of forage-fed beef steers

Although cationic amino acids (CAA) are considered essential to maximize optimal growth of cattle, transporters responsible for CAA absorption by bovine small intestinal epithelia have not been described. This study was conducted to test 2 hypotheses: 1) the duodenal, jejunal, and ileal epithelia of beef cattle differentially express 7 mRNA associated with 4 mammalian amino acid (AA) transport activities: y+ (CAT1), B0,+ (ATB0,+), b0,+ (b0,+AT and rBAT), and y+L (y+LAT1, y+LAT2, and 4F2hc), and 2) the expression of these mRNA is responsive to small intestinal luminal supply of AA substrates (derived from ruminal microbes) or glucose-derived energy (from starch hydrolysate, SH), or both. Eighteen ruminally and abomasally catheterized Angus steers (body weight = 260±17kg) fed an alfalfa cube-based diet at 1.33×net energy for maintenance requirement were assigned to 3 treatments (n=6): ruminal and abomasal water infusion (control); ruminal SH and abomasal water infusion; and ruminal water and abomasal SH infusion. The dosage of SH infusion amounted to 20% of metabolizable energy intake. After 14 or 16 d of infusion, steers were slaughtered, duodenal, jejunal, and ileal epithelia were harvested, and total RNA was extracted. The relative amounts of mRNA expressed by epithelia were quantified using real-time reverse transcription-PCR. All 7 mRNA species were expressed by the epithelium from each region, but their abundance differed among the regions. Specifically, duodenal expression of CAT1 and ATB0,+ mRNA was greater than jejunal or ileal expression; ileal expression of b0,+AT, rBAT, and y+LAT1 mRNA was greater than jejunal or duodenal expression, whereas the expression of y+LAT2 and 4F2hc mRNA did not differ among the 3 epithelia. With regard to SH infusion effect, ruminal infusion down-regulated or tended to down-regulate the jejunal expression of CAT1, rBAT, y+LAT2, and 4F2hc mRNA. Abomasal infusion down-regulated the jejunal expression of y+LAT2 mRNA and tended to down-regulate the jejunal expression of 4F2hc mRNA. This study characterized the pattern of CAA transporter mRNA expressed by growing beef cattle fed an alfalfa-based diet. Moreover, this study demonstrated that increasing the luminal supply of microbe-derived AA (by ruminal supplementation of SH) results in a reduced capacity of apical and basolateral membrane to transport of CAA, whereas increasing luminal glucose supply (by abomasal supplementation of SH) reduces only the basolateral transport capacity, assuming that CAA transporter mRNA content represents functional capacity.

Responses in mammary and splanchnic metabolism to altered lysine supply in dairy cows

Lysine is usually taken up in excess by the mammary gland (MG) relative to milk protein output, allowing for mammary synthesis of non-essential (NE) amino acids (AA) from Lys-N. It is unclear whether this NEAA synthesis from Lys is obligate or whether more efficient use of Lys can be made under limiting conditions. Six multi-catheterized dairy cows received a basal diet low in protein plus an abomasal infusion of AA (560 g/day) with or without Lys (50.3 g/day), in a crossover design with 7-day periods. On day 7, all cows received a 7.5-h jugular infusion of [2-15N]lysine. Six blood samples were collected from arterial, portal, hepatic and mammary vessels at 45 min intervals. In addition, cows were milked at 6 and 7 h with the milk casein plus arterial and mammary plasma collected at 7 h analyzed for AA enrichment. Milk protein concentration and casein yield tended (P < 0.10) to decrease with Lys deletion, while Lys secretion in milk protein was lowered (P < 0.05). The addition of Lys in the AA mixture increased the net portal absorption of Lys by the amount infused, suggesting limited oxidation of this extra supply by the gut. Net liver flux of Lys was unaltered by treatment and, therefore, net splanchnic release of Lys reflected closely the amounts absorbed. For both treatments, however, post-liver supply was greater than mammary uptake, which exceeded milk output. Nonetheless, while Lys deletion decreased mammary uptake by 10.1 mmol/h, Lys in milk protein secretion was reduced by only 3.9 mmol/h. On a net basis, there was no evidence of the additional uptake of any other measured AA during the Lys deletion. The mammary uptake to output ratio of Lys decreased from 1.37 to 1.12, but still showed an excess with Lys deletion. The total amount of 15N in milk protein did not change with treatment but the distribution into AA was altered. In conditions that simulated normal feeding (Lys infused), 83% of the 15N was present as Lys, with Glx, Asx, Ser and Ala harvesting, respectively, 6.8%, 2.4%, 2.1% and 1.0%. With Lys depletion, N-transfers from Lys to other AA within the MG were still present, but rates were considerably lower. This would suggest that part, at least, of Lys catabolism in the MG is either needed or cannot be prevented completely, even at low supply of Lys. Such catabolism will provide N to support the synthesis of NEAA.

Uptake and Utilization of Trans Octadecenoic Acids in Lactating Dairy Cows

Trans fatty acids (FA) arise in ruminant-derived foods as a consequence of rumen biohydrogenation and are of interest because of their biological effects and potential role in chronic human diseases. Our objective was to compare 2 trans FA, elaidic acid (EA; trans-9 18:1) and vaccenic acid (VA; trans-11 18:1), with oleic acid (OA; cis-9 18:1) relative to plasma lipid transport and mammary utilization for milk fat synthesis. Three ruminally cannulated, Holstein dairy cows, 259±6 DIM (mean±SEM), were randomly assigned in a 3×3 Latin square design. Treatments were a 4-d abomasal infusion of 1) OA (45.5 g/d), 2) EA (41.7 g/d), and 3) VA (41.4 g/d). Milk samples were collected at each milking and blood samples were collected at the start and end of each treatment period. The proportions of total plasma FA associated with each plasma lipid fraction at baseline (pretreatment) were 62.6±0.6% phospholipids, 26.1±0.6% cholesterol esters, 9.8±0.4% triglycerides, and 1.5±0.1% nonesterified fatty acids; these values were unaffected by treatment. There were striking differences in the FA composition of the individual plasma lipid fractions and in the distribution of specific 18-carbon FA among the lipid fractions. Infusion of treatment isomers caused their specific increase in the various plasma lipid fractions but had no effect on milk production variables, including milk fat yield and content. Transfer efficiency of infused OA, EA, and VA to milk fat averaged 65.5±3.0%, 59.7±1.5%, and 54.3±0.6%, respectively. For the VA infusion, 24.6±1.1% of the transfer was accounted for by the increased yield of cis-9, trans-11 conjugated linoleic acid in milk fat, consistent with its endogenous synthesis from VA via the mammary enzyme Δ9-desaturase. Notably, linoleic acid (18:2n-6) and linolenic acid (18:3n-3) accounted for 47.7% of total plasma FA, but only 2.6% of FA in milk. Overall, results demonstrate clear differences in plasma transport and mammary uptake and utilization of 18-carbon FA, and these relate to the location, orientation, and number of double bonds.

Limiting amino acids for growing lambs fed a diet low in ruminally undegradable protein1

Ruminally cannulated Rambouillet wether lambs were used in three 6 x 6 Latin square experiments (n = 6/experiment) to determine which essential AA limit N retention. Lambs (BW = 36.9 +/- 1.9 kg for Exp. 1, 35.1 +/- 1.4 kg for Exp. 2, and 46.0 +/- 1.3 kg for Exp. 3) were housed in metabolism crates and limit-fed (DMI = approx. 1.8% of BW daily) twice daily a soybean hull-based diet low in ruminally undegradable protein. Treatments for Exp. 1 were continuous abomasal infusions of a solution (500 mL/d) containing 1) no AA (CON), 2) a mixture of 10 essential AA and 2 nonessential AA (10EAA), 3) 10EAA with Met removed, 4) 10EAA with Lys removed, 5) 10EAA with His removed, and 6) 10EAA with Thr removed. Treatments for Exp. 2 were abomasal infusions of 1) CON, 2) 10EAA, 3) 10EAA with Leu, Ile, and Val removed (-BCAA), 4) 10EAA with Arg removed, 5) 10EAA with Phe removed, and 6) 10EAA with Trp removed. Treatments for Exp. 3 were abomasal infusions of 1) CON, 2) 10EAA, 3) -BCAA, 4) 10EAA with Leu removed, 5) 10EAA with Ile removed, and 6) 10EAA with Val removed. All lambs received continuous infusions of acetate and propionate into the rumen and dextrose into the abomasum to supply additional energy. Periods were 7 d: 3 d for adaptation to abomasally infused treatments and 4 d for fecal and urinary collections. Blood samples were collected 3 h after feeding on d 7. In all 3 experiments, N retention was greater (P < 0.10) for lambs receiving 10EAA vs. CON, demonstrating that the basal AA supply from CON was limiting. Removal of each of the essential AA from 10EAA decreased (P < 0.10) their concentrations in plasma (except for Trp), indicating that 10EAA supplied these AA in excess of the animal's requirement. In Exp. 1, N retention (g/d) decreased (P < 0.10) in response to the removal of Met and Thr, but was not affected by removal of Lys and His from 10EAA. In Exp. 2, N retention decreased (P < 0.10) in response to removal of all 3 branched-chain AA, Arg, and Trp, whereas the removal of Phe from 10EAA did not affect N retention. In Exp. 3, N retention decreased (P < 0.10) in response to removal of branched-chain AA and Val, but was not affected by the omission of Leu and Ile from 10EAA. The results of this research demonstrated that Met, Thr, Arg, Trp, and Val limited N retention of lambs fed a diet low in ruminally undegradable protein.