Morphologic and transcriptomic assessment of bovine embryos exposed to dietary long-chain fatty acids.

The main objectives of this study were to determine the influence of diets enriched in α-linolenic, linoleic or oleic acid on the development and transcriptomic profile of embryos collected from dairy cattle. Non-lactating Holstein cows received one of the three diets supplemented with 8% rolled oilseeds: flax (FLX, n = 8), sunflower (SUN, n = 7) or canola (CAN, n = 8). After a minimum 35-day diet adaptation, cows were superovulated, artificially inseminated and ova/embryos recovered non-surgically after 7.5 days. Cows fed FLX had less degenerated embryos and more viable embryos than those fed CAN or SUN. In total, 175 genes were differentially expressed in blastocysts from cows fed FLX than in cows fed CAN or SUN. These differentially expressed genes were mainly involved in cellular growth and proliferation, cellular development, and cell survival and viability. In conclusion, dietary n-3 polyunsaturated fatty acids reduced early embryonic degeneration possibly through improving embryonic cell survival and viability.

A prepartum diet supplemented with oilseeds high in oleic or linoleic acid reduced GnRH-induced LH release in dairy cows during second week postpartum.

BACKGROUND: The objective was to determine the effect of prepartum diets supplemented with rolled canola seed (high in oleic acid) or sunflower seed (high in linoleic acid) on luteinizing hormone (LH) pulsatility and gonadotropin releasing hormone (GnRH)-induced LH release during early postpartum. METHODS: Thirty-one pregnant Holstein cows, blocked by body condition score, parity and expected calving date, were assigned to 1 of 3 prepartum diets supplemented with 8 % rolled canola or sunflower seed, or no oilseed (control) during the last 35 d of gestation. Blood samples were collected at Weeks (wk)-3, 0, +1 and +2, relative to calving, to determine non-esterified fatty acids (NEFA), Beta-hydroxy butyric acid (BHBA) and glucose. Additional blood samples were collected during wk1 (n = 5 per treatment) or wk2 (n = 5 or 6 per treatment), for 6 h, to measure LH pulsatility; thereafter, 100 mcg GnRH was administrated i.m., and blood was sampled for 4 h more, to measure GnRH-induced LH release. RESULTS: Dietary treatment did not affect prepartum energy balance, but cows fed the control diet were in a deeper state of negative energy balance during wk2, than those fed canola (P = 0.03) or sunflower (P = 0.01). Prepartum diets did not influence the mean plasma concentration of BHBA and glucose. However, NEFA concentration during wk2 was greater in control cows than those fed sunflower (P = 0.03) or canola (P = 0.07). Prepartum diets did not affect LH pulsatility (i.e. mean, minimum, maximum concentration, pulse frequency, and amplitude during wk1 and 2). GnRH-induced LH release did not differ among dietary treatments during wk1 but the mean GnRH-induced LH release during wk2 was either greater (P = 0.02) and tended to be greater (P = 0.09) in control cows than in those fed canola and sunflower, respectively. CONCLUSIONS: Prepartum diets did not affect LH pulsatility and GnRH-induced LH release during the first week postpartum, but cows fed a diet supplemented with oilseeds high in oleic or linoleic acid released less LH than control cows, in response to an exogenous GnRH challenge during the second week postpartum.

Effect of calf starter feeding on gut microbial diversity and expression of genes involved in host immune responses and tight junctions in dairy calves during weaning transition.

Calf starters are usually offered to dairy calves to facilitate the weaning process, however, the effect of solid feed consumption on gut health has not been well studied. This study aimed to investigate the effect of calf starter feeding on the gut bacterial community and mucosal immune functions in dairy calves during weaning transition. Mucosal tissue and digesta samples were collected from rumen, jejunum, ileum, cecum, and colon upon slaughtering of calves (n=8) after feeding the experimental diets [milk replacer (MR) or milk replacer + calf starter (MR+S)] for 6 wk. Expression of toll-like receptor (TLR) 10 was downregulated along the gut, whereas TLR2 in colon and TLR6 along the gut were upregulated in MR+S-fed calves compared with MR-fed calves. Ileal TLR9 and TLR10 showed higher expression compared with the other regions regardless of the diet. Peptidoglycan recognition protein 1 demonstrated a diet- and gut-regional dependent expression pattern, whereas ß-defensin did not. The diet and gut region also affected the expression of tight junction-regulating genes claudin 4 and occludin. Bacterial diversity tended to be different between the 2 diets, whereas the bacterial density was different among gut regions and sample type. The present study revealed that changes in bacterial diversity, expression of genes encoding host mucosal immune responses, and barrier functions were associated with the MR+S diet, and suggests that solid feed consumption may alter gut microbiome and host mucosal functions during weaning transition.

Changes in bacterial diversity associated with epithelial tissue in the beef cow rumen during the transition to a high-grain diet.

Our understanding of the ruminal epithelial tissue-associated bacterial (defined as epimural bacteria in this study) community is limited. In this study, we aimed to determine whether diet influences the diversity of the epimural bacterial community in the bovine rumen. Twenty-four beef heifers were randomly assigned to either a rapid grain adaptation (RGA) treatment (n = 18) in which the heifers were allowed to adapt from a diet containing 97% hay to a diet containing 8% hay over 29 days or to the control group (n = 6), which was fed 97% hay. Rumen papillae were collected when the heifers were fed 97%, 25%, and 8% hay diets. PCR-denaturing gradient gel electrophoresis (DGGE) and quantitative real-time PCR analysis were used to characterize rumen epimural bacterial diversity and to estimate the total epimural bacterial population (copy numbers of the 16S rRNA gene). The epimural bacterial diversity from RGA heifers changed (P = 0.01) in response to the rapid dietary transition, whereas it was not affected in control heifers. A total of 88 PCR-DGGE bands were detected, and 44 were identified from phyla including Firmicutes, Bacteroidetes, and Proteobacteria. The bacteria Treponema sp., Ruminobacter sp., and Lachnospiraceae sp. were detected only when heifers were fed 25% and 8% hay diets, suggesting the presence of these bacteria is the result of adaptation to the high-grain diets. In addition, the total estimated population of rumen epimural bacteria was positively correlated with molar proportions of acetate, isobutyrate, and isovalerate, suggesting that they may play a role in volatile fatty acid metabolism in the rumen.

Serum paraoxonase-1 activity in tail and mammary veins of ketotic dairy cows.

The objective of this study was to evaluate the association between ketonemia and serum paraoxonase-1 (PON1), malondialdehyde (MDA), and other blood components in tail and mammary veins of dairy cows. Forty-two Holstein dairy cows with decreased feed intake were divided into HIGH (≥ 1.2 mM; n = 31) and LOW (< 1.2 mM; n = 11) groups based on the ß-hydroxybutyrate concentration in plasma collected from the tail vein. The HIGH group had a significantly greater plasma non-esterified fatty acid (NEFA) concentration, but significantly lower serum PON1 activity and phospholipid concentration, and a tendency to have a lower cholesterol ester concentration than the LOW group. Serum PON1 activity was not correlated with the MDA concentration but was positively correlated with serum concentrations of cholesterol esters and phospholipids, and negatively correlated with the plasma NEFA concentration. These results suggest that serum PON1 activity is reduced by hyperketonemia and the relevance of PON1 to MDA seems to not be direct, though it is involved.

L'objectif de la présente étude était d'évaluer l'association entre l'acétonémie et la paraoxonase-1 (PON1), le malondialdéhyde (MDA), et d'autres composés du sang dans les veines caudale et mammaire de vaches laitières. Quarante-deux vaches laitières de race Holstein présentant une diminution de l'ingestion d'aliments furent divisées en groupes ÉLEVÉ (≥ 1,2 mM; n = 31) et BAS (< 1,2 mM; n = 11) basés sur la concentration de ß-hydroxybutyrate de plasma prélevé de la veine caudale. Le groupe ÉLEVÉ avait une concentration plasmatique significativement plus grande d'acides gras non-estérifiés (NEFA), mais le sérum présentait une activité PON1 et une concentration de phospholipides significativement réduite, et une tendance à avoir une concentration d'esters de cholestérol plus faible que le groupe BAS. L'activité de PON1 sérique n'était pas corrélée avec la concentration de MDA mais était corrélée positivement avec les concentrations sériques d'esters de cholestérol et de phospholipides, et corrélée négativement avec la concentration plasmatique de NEFA. Ces résultats suggèrent que l'activité de PON1 sérique est réduite par l'hypercétonémie et la pertinence de PON1 envers MDA ne semble pas être directe, bien qu'elle semble impliquée.(Traduit par Docteur Serge Messier).

Epithelial capacity for apical uptake of short chain fatty acids is a key determinant for intraruminal pH and the susceptibility to subacute ruminal acidosis in sheep.

Subacute ruminal acidosis (SARA) is a common digestive disorder occurring in ruminants, with considerable variation in the severity of SARA observed among animals fed the same diet. Our aim in this study was to determine whether differences in the capacity of the ruminal epithelium for the apical uptake of acetate and butyrate (determined in Ussing chambers after slaughter) explains differences observed for the severity of a preceding episode of SARA in vivo. Adult sheep with an indwelling small ruminant ruminal pH measurement system (SRS) were randomly assigned to either a SARA induction treatment (oral drench containing 5 g glucose/kg body weight; n = 17) or a sham treatment (SHAM; n = 7; 12 mL water/kg body weight). Sheep receiving the glucose drench were further classified as nonresponders (NR; n = 7) or responders (RES; n = 7) according to their ruminal pH profile for the 3 h following the oral drench. Mean ruminal pH for the 3 h following the drench differed among groups (P < 0.001), with it being highest for SHAM (6.67 +/- 0.08), intermediate for NR (5.97 +/- 0.05), and lowest for RES (5.57 +/- 0.08) sheep. The apical uptake of acetate and butyrate did not differ between SHAM and RES sheep. However, NR sheep had greater in vitro apical uptake of acetate and butyrate and a higher plasma beta-hydroxybutyrate concentration than RES sheep, suggesting greater absorptive capacity for NR. Differences between NR and RES were attributed to greater bicarbonate-independent, nitrate-sensitive uptake of acetate (P = 0.007), a tendency for greater bicarbonate-dependent uptake of acetate (P = 0.071), and greater bicarbonate-independent uptake of butyrate (P = 0.022). These data indicate that differences in the rates and pathways for the uptake of acetate and butyrate explain a large proportion of the individual variation observed for the severity of SARA.

Commensal fecal Escherichia coli diversity in dairy cows at high and low risk for incurring subacute ruminal acidosis.

Subacute ruminal acidosis (SARA) is a common digestive disorder in dairy cows characterized by prolonged periods of undesirably low rumen pH (<5.8) and is caused by the accumulation of volatile fatty acids in rumen. This disorder damages the ruminal mucosa, causes diarrhea, reduces dry matter intake (DMI), and can result in anorexia and death. In this study, nonlactating dairy cows were fed diets predisposing them to a high risk (HR; n = 6) or a low risk (LR; n = 6) for experiencing SARA. The goal was to investigate differences in antimicrobial resistance selection, proliferation, and characterization of Escherichia coli strain types among the two treatment groups. Fecal samples were used to isolate total, tetracycline-resistant (Tet(r)), and ampicillin-resistant E. coli, and selected isolates were examined. We found reduced total (1.2-fold) and Tet(r) (1.4-fold) E. coli in HR cows. Low ampicillin-resistant E. coli shedding was detected from both HR (0.22 colony forming unit/g) and LR (0.46 colony forming unit/g) cows. Overall, 39 pulsed-field gel electrophoresis (PFGE) profiles and 13 antibiotic resistance profiles (phenotypes) were identified from the total isolates examined (n = 144). The LR cows exhibited diverse genotypes (22 PFGE profiles) clustering into seven restriction endonuclease digestion pattern clusters (REPCs) within total and Tet(r) E. coli. In comparison, isolates from HR animals showed increased genotypic relatedness (16 PFGE profiles and 13 REPC with comparable phenotypes). From both HR and LR cows, no significant differences in the detection of a particular phenotype were observed (p > 0.05), and tet(A) allele was frequently detected among isolates from HR (45.2%) and tet(B) from LR (36.6%) cows. Changes in fecal E. coli genotypes should be explored further for its usefulness as an indicator for SARA since dairy cows are a reservoir of diverse E. coli strain types. Our results elucidate phenotypic and genotypic differences in fecal E. coli shed between HR and LR cows.

Use of gallic acid and hydrolyzable tannins to reduce methane emission and nitrogen excretion in beef cattle fed a diet containing alfalfa silage1,2.

The objective of this study was to determine the effects of different forms of hydrolyzable tannin [HT; source (chestnut, CN; tannic acid, TA); subunit (gallic acid, GA)] on apparent total-tract digestibility, methane (CH4) production, and nitrogen (N) utilization in beef cattle fed an alfalfa silage-based diet. Eight ruminally cannulated heifers with an initial BW of 480 ± 29.2 kg (mean ± SD) were used in a double 4 × 4 Latin square experiment. The experiment consisted of four 28-d periods (14-d adaptation, 14-d measurements) and a 7-d washout between periods. The animals received a basal diet with 19.8% CP (DM basis) content containing 75% alfalfa silage, 20% barley silage, and 5% supplement (DM basis) with or without different forms of HT. The dietary treatments were as follows: control (no HT), GA (1.5% of diet DM), TA (1.5% of diet DM), and CN (2% of diet DM). Animals were fed 95% of their ad libitum intake during the measurement phase. Total fecal excretion was collected for 4 d, CH4 was measured for 72 h using respiration chambers, and ruminal fermentation variables and plasma urea N (PUN) concentration were measured on 2 nonconsecutive days before and after feeding. The restricted DM (DMI; 10.79 ± 1.076 kg/d) and nutrient intakes did not differ (P ≥ 0.22) among treatments. Furthermore, apparent DM digestibility (60.3 ± 0.86%) was not affected (P = 0.20) by treatment, but CP digestibility decreased for TA and CN compared with control and GA treatments (63.1 vs. 69.0%; P < 0.001). Total VFA concentration tended (P = 0.089) to increase for GA compared with control and TA (134 vs. 125 and 126 mM) and intermediate for CN (129 mM). The PUN concentration was lower for all HT treatments compared with control (196 vs. 213 mg/L; P = 0.02). Both TA and CN increased the proportion of N excreted in feces and decreased the proportion in urine compared with control and GA (43.9% vs. 37.8% and 56.1% vs. 62.2%; respectively; P < 0.001). However, the proportion of urea N in urinary N decreased for all HT treatments compared with control (47.2% vs. 51.2%; P = 0.02). Also, GA tended to decrease CH4/DMI (20.4 vs. 22.3 g/kg DMI; P = 0.07) and decreased the proportion of GE intake emitted as CH4 (5.16 vs. 5.71%; P = 0.04) compared with control. Thus, among the different forms of HT applied to a high-protein alfalfa silage-based diet, both TA and CN had no effect on CH4 production, but decreased CP digestibility and shifted N excretion from urine to feces, whereas GA (i.e., HT subunit) decreased CH4 production and decreased the proportion of urea N in urinary N in beef cattle without affecting CP digestibility. Thus, feeding the HT subunit, GA, has the potential to decrease environment impact of ruminants (lower CH4 and ammonia emissions), without decreasing animal performance.

Effects of hydrolyzable tannin with or without condensed tannin on methane emissions, nitrogen use, and performance of beef cattle fed a high-forage diet.

Sustainability of animal agriculture requires efficient use of energy and nitrogen (N) by ruminants fed high-forage diets. Thus, there is a need to decrease methane (CH4) emissions and prevent excessive N release into the environment. Therefore, this experiment examined the long-term effects of feeding hydrolyzable tannin (HT) with or without condensed tannin (CT) on animal performance, rumen fermentation, N use, and CH4 production in beef cattle fed a high-forage diet. A total of 75 weaned crossbred steers (292 ± 4.1 kg) were grouped by body weight (BW), housed in individual pens, and randomly assigned to 1 of 5 dietary treatments (15 animals/treatment) in a completely random design. The animals were fed a basal diet of alfalfa:barley silages (50:50; dry matter [DM] basis) with a crude protein content of 17.1% and supplemented with HT extract (chestnut, CN) or a combination (50:50) of HT and CT extracts (quebracho, Q) in a powdered form at different levels of dietary DM. The treatments for determining animal performance and N use were control (no tannin), 0.25% CN, 1.5% CN, combination of CN and Q at 0.125% each (0.25% CNQ), and CN and Q at 0.75% each (1.5% CNQ) of dietary DM. The treatments for the CH4 measurement were control, 1.5% CN, and 1.5% CNQ of dietary DM. The first 84 d of the study were used to measure animal performance, rumen fermentation, and N use, and the next 30 d were used to measure CH4 emissions with the tracer gas technique. There were no effects of treatment on DM intake (DMI), BW, average daily gain, and gain: feed (P ≥ 0.10). The plasma urea N concentration was greater (P < 0.05) for 1.5% CN and 1.5% CNQ than those fed 0.25% CNQ (120.9 and 120.4 vs. 111.7 mg/L, respectively), but not different (P > 0.05) from animals fed control or 0.25% CN (117.2 and 117.5 mg/L, respectively). Tannin inclusion did not affect rumen pH, total volatile fatty acid concentration, proportions of acetate and propionate, and total protozoa populations (P ≥ 0.16). Tannin, irrespective of type or dose, decreased (P < 0.01) ruminal ammonia concentration. Tannin type and dose did not affect (P = 0.54) daily CH4 production (154 ± 5.9 g/d) but 1.5% CNQ tended to decrease CH4 yield compared with control (20.6 vs. 22.0 g/kg DMI; P = 0.094). HT from CN alone or in combination with CT from Q can be added at a low (0.25% DM) or high (1.5% DM) level to a forage-based diet to decrease ruminal ammonia concentration in growing beef cattle fed a high-protein diet without adverse effects on animal performance. A combination of HT and CT at a concentration of 1.5% dietary DM also tended to decrease CH4 emissions without negatively affecting performance.

Assessment of microbiome changes after rumen transfaunation: implications on improving feed efficiency in beef cattle.

BACKGROUND: Understanding the host impact on its symbiotic microbiota is important in redirecting the rumen microbiota and thus improving animal performance. The current study aimed to understand how rumen microbiota were altered and re-established after being emptied and receiving content from donor, thus to understand the impact of such process on rumen microbial fermentation and to explore the microbial phylotypes with higher manipulation potentials. RESULTS: Individual animal had strong effect on the re-establishment of the bacterial community according to the observed profiles detected by both fingerprinting and pyrosequencing. Most of the bacterial profile recovery patterns and extents at genus level varied among steers; and each identified bacterial genus responded to transfaunation differently within each host. Coriobacteriaceae, Coprococcus, and Lactobacillus were found to be the most responsive and tunable genera by exchanging rumen content. Besides, the association of 18 bacterial phylotypes with host fermentation parameters suggest that these phylotypes should also be considered as the regulating targets in improving host feed efficiency. In addition, the archaeal community had different re-establishment patterns for each host as determined by fingerprint profiling: it was altered after receiving non-native microbiome in some animals, while it resumed its original status after the adaptation period in the other ones. CONCLUSIONS: The highly individualized microbial re-establishment process suggested the importance of considering host genetics, microbial functional genomics, and host fermentation/performance assessment when developing effective and selective microbial manipulation methods for improving animal feed efficiency.

Propionate is not an important regulator of plasma leptin concentration in dairy cattle.

Propionate was recently shown to increase leptin synthesis in rodents. To determine if a similar effect occurs in ruminants, propionate was administered to lactating dairy cows. In experiment 1, 31 cows were given an intrajugular Na propionate bolus (1,040 micromol/kg body weight), increasing plasma propionate from 160 to 5,680 microM and plasma insulin from 6.8 to 77.8 microIU/mL. Plasma leptin concentration decreased from 2.11 ng/mL before bolus to 1.99 ng/mL after dosing (P<0.05) with no differences in leptin concentrations at 20, 50, and 100 min post-bolus (P>0.10). In experiment 2, 12 cows were used in a duplicated 6 x 6 Latin square experiment to assess the dose-response effect of ruminal propionate infusion on plasma leptin concentration. Sodium propionate was infused at rates of 0, 260, 520, 780, 1040, or 1,300 mmol/h, while total short-chain fatty acid infusion rate was held constant at 1,300 mmol/h by addition of Na acetate to the infusate. Coccygeal blood was sampled following 18 h of infusion. Increasing the rate of propionate infusion linearly increased plasma propionate concentration from 180 to 330 microM (P<0.001) and plasma insulin concentration from 6.7 to 9.1 microIU/mL (P<0.05). There was a quadratic response in plasma leptin concentration (P=0.04) with a maximum at 780 mmol/h propionate, but leptin concentrations increased by no more than 8% relative to the 0 mmol/h propionate infusion. Leptin concentrations were correlated with insulin concentrations but not with propionate concentrations in plasma. Propionate is not a physiological regulator of leptin secretion in lactating dairy cows.

Fermentation of Ammonia Fiber Expansion Treated and Untreated Barley Straw in a Rumen Simulation Technique Using Rumen Inoculum from Cattle with Slow versus Fast Rate of Fiber Disappearance.

The purpose of this study was to determine the effect of rumen inoculum from heifers with fast vs. slow rate of in situ fiber digestion on the fermentation of complex versus easily digested fiber sources in the forms of untreated and Ammonia Fiber Expansion (AFEX) treated barley straw, respectively, using an artificial rumen simulation technique (Rusitec). In situ fiber digestion was measured in a previous study by incubating untreated barley straw in the rumen of 16 heifers fed a diet consisting of 700 g/kg barley straw and 300 g/kg concentrate. The two heifers with fastest rate of digestion (Fast ≥ 4.18% h-1) and the two heifers with the slowest rate of digestion (Slow ≤ 3.17% h-1) were chosen as inoculum donors for this study. Two Rusitec apparatuses each equipped with eight fermenters were used in a completely randomized block design with two blocks (apparatus) and four treatments in a 2 × 2 factorial arrangement of treatments (Fast or Slow rumen inoculum and untreated or AFEX treated straw). Fast rumen inoculum and AFEX straw both increased (P < 0.05) disappearance of dry matter (DMD), organic matter, true DMD, neutral detergent fiber, acid detergent fiber, and nitrogen (N) with an interactive effect between the two (P < 0.05). Fast rumen inoculum increased (P > 0.05) methane production per gram of digested material for both untreated and AFEX straw, and reduced (interaction, P < 0.05) acetate: propionate ratio for untreated straw. Greater relative populations of Ruminococcus albus (P < 0.05) and increased microbial N production (P = 0.045) were observed in Fast rumen inoculum. AFEX straw in Fast inoculum had greater total bacterial populations than Slow, but for untreated straw this result was reversed (interaction, P = 0.013). These findings indicate that differences in microbial populations in rumen fluid contribute to differences in the capacity of rumen inoculum to digest fiber.

Variation of bacterial communities and expression of Toll-like receptor genes in the rumen of steers differing in susceptibility to subacute ruminal acidosis.

In order to determine differences in the ruminal bacterial community and host Toll-like receptor (TLR) gene expression of beef cattle with different susceptibility to acidosis, rumen papillae and content were collected from acidosis-susceptible (AS, n=3) and acidosis-resistant (AR, n=3) steers. The ruminal bacterial community was characterized using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and quantitative real time PCR (qRT-PCR) analysis. Global R analysis of bacterial profile similarity revealed that bacterial diversity was significantly different between AR and AS groups for both rumen content (P=0.001) and epithelial (P=0.002) communities. The copy number of total bacterial 16S rRNA genes in content of AS steers was 10-fold higher than that of AR steers, and the copy number of total 16S rRNA genes of epimural bacteria in AR steers was positively correlated with ruminal pH (r=0.59, P=0.04), and negatively correlated with total VFA concentration (r=-0.59, P=0.05). The expressions of host TLR2 and 4 genes were significantly higher in AR steers compared to those in AS steers. These findings enhance our understanding about the ruminal microbial ecology and host gene expression changes that may be useful in the prevention of ruminal acidosis.

Comparative analysis of gene expression profiles in ruminal tissue from Holstein dairy cows fed high or low concentrate diets.

Cattle are often fed high concentrate (HC) diets to increase productivity, although HC diets cause changes in ruminal environment such as pH reduction. Despite those well-documented changes in cattle fed HC diets, there is currently a paucity of data describing the molecular events regulating the ruminal environment. Our objective was to gain an understanding of which genes are differentially expressed in ruminal tissue from Holstein cows fed a HC comparing to low concentrate (LC) diet using microarray analysis using a bovine 24 k microarray. A total of 5,200 differentially expressed genes (DEG) were detected for cows fed HC relative to LC. The DEG were firstly annotated with gene ontology (GO) and Kyoto encyclopedia of Genes and Genomes (KEGG), indicating that the DEG were associated with catalytic activity and MAPK pathway, respectively. Further characterization using GeneCodis identified patterns of interrelated annotations for the DEG to elucidate the relationships among annotation groups revealed that a cAMP-dependent protein kinase A catalytic subunit beta (PRKACB), may be associated with ruminal tissue maintenance. The results contributed to understanding of the regulatory mechanisms at the mRNA level for Holstein cows fed at different concentrate ratio diets.

Fecal and urinary lignans, intrafollicular estradiol, and endometrial receptors in lactating dairy cows fed diets supplemented with hydrogenated animal fat, flaxseed or sunflower seed.

We hypothesized that the inclusion of flaxseed in the diets of lactating dairy cows will increase urinary and fecal concentrations of the lignans, secoisolariciresinol diglycoside (SDG), enterolactone and enterodiol, reduce intrafollicular concentrations of IGF-I and estradiol, and subsequently reduce estradiol and oxytocin receptor expression in the endometrium. To test this hypothesis, 27 cycling, lactating Holstein cows were assigned to 1 of 3 diets supplemented with saturated fatty acids (SAT), flax (FLX), or sunflower (SUN) seed. Rations were formulated to provide 750 g supplemental fat/cow/d in all dietary groups. Ovulation (Day 0) was synchronized, and 5 d later, follicles>8 mm were ablated by an ultrasound-guided procedure in all cows. Samples of blood (Days 0 to 14), follicular fluid (Day 5 and 15), endometrium (Day 15), as well as urine and feces were collected in a subset of the animals. The fecal concentrations of SDG and enterodiol were higher (P<0.05) in cows fed FLX than in those fed SAT or SUN. Enterodiol increased (P<0.05) in urine samples of cows fed FLX, compared to those of cows fed SUN. However, follicular estradiol concentrations on Day 5 and 15 and endometrial concentrations of estradiol and oxytocin receptors on Day 15 did not differ among the dietary groups. Mean plasma progesterone concentrations were higher (P<0.05) in cows fed FLX and SUN than in those fed SAT. In summary, a diet supplemented with flaxseed increased the concentrations of SDG and enterodiol in feces, as hypothesized, but did not alter intrafollicular concentrations of IGF-I or estradiol, or endometrial populations of oxytocin or estrogen receptors in lactating dairy cows.

Intraruminal infusion of propionate alters feeding behavior and decreases energy intake of lactating dairy cows.

The dose-response effects of intraruminal infusion of propionate on feeding behavior of lactating dairy cows were evaluated with eight ruminally cannulated Holstein cows past peak lactation. Treatments were mixtures of propionic acid and acetic acid containing propionic acid at eight different concentrations in Experiment 1, and mixtures of sodium propionate and sodium acetate containing sodium propionate at 4 different concentrations in Experiment 2. Experimental designs were an 8 x 8 and duplicated 4 x 4 Latin squares, respectively, for Experiments 1 and 2. Treatment solutions were infused into the rumen continuously for 14 h at a rate of 16.7 and 25 mmol/min, respectively, for Experiments 1 and 2. Infusion started 2 h before feeding and ended 12 h after feeding; feeding behavior was monitored for 12 h after feeding using a computerized data acquisition system. Total metabolizable energy (ME) intake was calculated by adding the energy of infusates to dietary energy intake. In both experiments, as the proportion of propionate of the infusates increased, total ME intake and dry matter intake decreased linearly. As infusion of propionate increased, meal size tended (P < 0.09) to decrease linearly and intermeal interval tended (P < 0.07) to increase linearly in Experiment 1; meal size decreased linearly and number of meal bouts tended (P < 0.08) to decrease linearly in Experiment 2. These observations indicate that the reduction in dietary energy intake from propionate infusion was greater than the energy supplied from infusates, and that propionate plays an important role in feed intake regulation by affecting both satiety and hunger.

Hypophagic effects of ammonium are greater when infused with propionate compared with acetate in lactating dairy cows.

The objective of this experiment was to determine interactions between hypophagic effects of propionate and ammonium in lactating dairy cows. Eight ruminally cannulated Holstein cows in mid-lactation were used in a duplicated 4 x 4 Latin square design experiment with a 2 x 2 factorial arrangement of treatments. Factors evaluated were type of volatile fatty acid (VFA; acetate vs. propionate) and type of salt (sodium vs. ammonium). Treatment solutions were infused continuously into the rumen at a rate of 16.7 mmol of VFA salts/min starting 2 h before feeding and ending 12 h after feeding. Hypophagic effects of ammonium were significantly greater for cows infused with propionate (4.3 vs. 12.1 kg/12 h; SEM = 1.3) compared with acetate (13.5 vs. 15.3 kg/12 h; SEM = 1.3; interaction P < 0.01). This interaction was attributed to a greater reduction in meal frequency for ammonium treatment compared with sodium treatment when infused with propionate (3.9 vs. 7.2/12 h; SEM = 0.8) compared with acetate (6.6 vs. 7.0/12 h; SEM = 0.8), indicating that infusion of ammonium propionate reduced hunger. Meal size was decreased by infusion of propionate compared with acetate, but was not affected by ammonium compared with sodium, indicating that ammonium did not affect satiety. Mechanisms to explain the interactions between ammonium and propionate on meal frequency and feed intake warrant further investigation.

Extent of hypophagia caused by propionate infusion is related to plasma glucose concentration in lactating dairy cows.

Two experiments were conducted to evaluate how dose-response effects of intraruminal infusion of propionate on feeding behavior and plasma metabolites are altered by diets differing in fermentability. Twelve ruminally cannulated Holstein cows were used in each experiment. Cows were fed diets containing either steam flaked corn or dry cracked corn (30% of dietary dry matter) in expt. 1, and diets differing in forage-to-concentrate ratio (66:34 vs. 36:64) in expt. 2. For both experiments, the experimental design was a crossover for dietary treatment, and a 6 x 6 Latin square for infusion treatment within a diet for each period. Infusion treatments were mixtures of sodium propionate and sodium acetate, containing propionate at 0, 0.2, 0.4, 0.6, 0.8 and 1.0 as a fraction of total volatile fatty acids infused. Treatment solutions were infused into the rumen continuously for 18 h starting 6 h before feeding at a rate of 23.1 mmol/min. Although propionate production from ruminal fermentation was expected to be different, dietary treatments did not affect dry matter intake (DMI) responses to propionate infusion for either experiment. However, propionate infusion decreased DMI linearly in expt. 1, but did not decrease DMI at lower rates of propionate infusion, which were much more effective at increasing plasma glucose concentration in expt. 2. Propionate had a smaller hypophagic effect at low concentrations of plasma glucose and had a greater hypophagic effect at elevated concentrations of plasma glucose, which could be explained by changes in the metabolism of propionate in the liver.