Evaluation of feed restriction and abomasal infusion of resistant starch as models to induce intestinal barrier dysfunction in healthy lactating cows.

Intestinal hyperpermeability and subsequent immune activation alters nutrient partitioning and thus, decreases productivity. Developing experimental models of intestinal barrier dysfunction in heathy cows is a prerequisite in identifying nutritional strategies to mitigate it. Six cannulated Holstein cows (mean ± standard deviation, 37 ± 10 kg/d milk yield; 219 ± 97 d in milk; 691 ± 70 kg body weight) were used in a replicated 3 × 3 Latin square design experiment with 21-d periods (16-d wash-out and 5-d challenge) to evaluate either feed restriction or hindgut acidosis as potential models for inducing intestinal hyperpermeability. Cows were randomly assigned to treatment sequence within square and treatment sequences were balanced for carryover effects. Treatments during the challenge were (1) control (CTR; ad libitum feeding); (2) feed restriction (FR; total mixed ration fed at 50% of ad libitum feed intake); and (3) resistant starch (RS; 500 g of resistant starch infused in abomasum once a day as a pulse-dose 30 min before morning feeding). The RS (ActiStar RT 75330, Cargill Inc.) was tapioca starch that was expected to be resistant to enzymatic digestion in the small intestine and highly fermentable in the hindgut. Blood samples were collected 4 h after feeding on d 13 and 14 of the wash-out periods (baseline data used as covariate), and on d 1, 3, and 5 of the challenge periods. Fecal samples were collected 4 and 8 h after the morning feeding on d 14 of the wash-out periods and d 5 of the challenge periods. By design, FR decreased dry matter intake (48%) relative to CTR and RS, and this resulted in marked reductions in milk and 3.5% FCM yields over time, with the most pronounced decrease occurring on d 5 of the challenge (34 and 27%, respectively). Further, FR increased somatic cell count by 115% on d 5 of the challenge relative to CTR and RS. Overall, FR increased nonesterified fatty acids (159 vs. 79 mEq/L) and decreased BHB (8.5 vs. 11.2 mg/dL), but did not change circulating glucose relative to CTR. However, RS had no effect on production or metabolism metrics. Resistant starch decreased fecal pH 8 h after the morning feeding (6.26 vs. 6.81) relative to CTR and FR. Further, RS increased circulating lipopolysaccharide binding protein (4.26 vs. 2.74 µg/mL) compared with FR only on d 1 of the challenge. Resistant starch also increased Hp (1.52 vs. 0.48 µg/mL) compared with CTR, but only on d 5 of the challenge. However, neither RS or FR affected concentrations of serum amyloid A, IL1ß, or circulating endotoxin compared with CTR. The lack of consistent responses in inflammatory biomarkers suggests that FR and RS did not meaningfully affect intestinal barrier function. Thus, future research evaluating the effects of hindgut acidosis and FR using more intense insults and direct metrics of intestinal barrier function is warranted.

Symposium review: The impact of absorbed nutrients on energy partitioning throughout lactation.

Most nutrition models and some nutritionists view ration formulation as accounting transactions to match nutrient supplies with nutrient requirements. However, diet and stage of lactation interact to alter the partitioning of nutrients toward milk and body reserves, which, in turn, alters requirements. Fermentation and digestion of diet components determine feeding behavior and the temporal pattern and profile of absorbed nutrients. The pattern and profile, in turn, alter hormonal signals, tissue responsiveness to hormones, and mammary metabolism to affect milk synthesis and energy partitioning differently depending on the physiological state of the cow. In the fresh period (first 2 to 3 wk postpartum), plasma insulin concentration and insulin sensitivity of tissues are low, so absorbed nutrients and body reserves are partitioned toward milk synthesis. As lactation progresses, insulin secretion and sensitivity increase, favoring deposition instead of mobilization of body reserves. High-starch diets increase ruminal propionate production, the flow of gluconeogenic precursors to the liver, and blood insulin concentrations. During early lactation, the glucose produced will preferentially be used by the mammary gland for milk production. As lactation progresses and milk yield decreases, glucose will increasingly stimulate repletion of body reserves. Diets with less starch and more digestible fiber increase ruminal production of acetate relative to propionate and, because acetate is less insulinogenic than propionate, these diets can minimize body weight gain. High dietary starch concentration and fermentability can also induce milk fat depression by increasing the production of biohydrogenation intermediates that inhibit milk fat synthesis and thus favor energy partitioning away from the mammary gland. Supplemental fatty acids also impact energy partitioning by affecting insulin concentration and insulin sensitivity of tissues. Depending on profile, physiological state, and interactions with other nutrients, supplemental fatty acids might increase milk yield at the expense of body reserves or partition energy to body reserves at the expense of milk yield. Supplemental protein or AA also can increase milk production but there is little evidence that dietary protein directly alters whole-body partitioning. Understanding the biology of these interactions can help nutritionists better formulate diets for cows at various stages of lactation.

Effects of hindgut acidosis on metabolism, inflammation, and production in dairy cows consuming a standard lactation diet.

Postruminal intestinal barrier dysfunction caused by excessive hindgut fermentation may be a source of peripheral inflammation in dairy cattle. Therefore, the study objectives were to evaluate the effects of isolated hindgut acidosis on metabolism, inflammation, and production in lactating dairy cows. Five rumen-cannulated lactating Holstein cows (32.6 ± 7.2 kg/d of milk yield, 242 ± 108 d in milk; 642 ± 99 kg of body weight; 1.8 ± 1.0 parity) were enrolled in a study with 2 experimental periods (P). During P1 (4 d), cows were fed ad libitum a standard lactating cow diet (26% starch dry matter) and baseline data were collected. During P2 (7 d), all cows were fed the same diet ad libitum and abomasally infused with 4 kg/d of pure corn starch (1 kg of corn starch + 1.25 L of H2O/infusion at 0600, 1200, 1800, and 0000 h). Effects of time (hour relative to the first infusion or day) relative to P1 were evaluated using PROC MIXED in SAS (version 9.4; SAS Institute Inc.). Infusing starch markedly reduced fecal pH (5.84 vs. 6.76) and increased fecal starch (2.2 to 9.6% of dry matter) relative to baseline. During P2, milk yield, milk components, energy-corrected milk yield, and voluntary dry matter intake remained unchanged. At 14 h, plasma insulin and ß-hydroxybutyrate increased (2.4-fold and 53%, respectively), whereas circulating glucose concentrations remained unaltered. Furthermore, blood urea nitrogen increased at 2 h (23%) before promptly decreasing below baseline at 14 h (13%). Nonesterified fatty acids tended to decrease from 2 to 26 h (40%). Circulating white blood cells and neutrophils increased on d 4 (36 and 73%, respectively) and somatic cell count increased on d 5 (4.8-fold). However, circulating serum amyloid A and lipopolysaccharide-binding protein concentrations were unaffected by starch infusions. Despite minor changes in postabsorptive energetics and leukocyte dynamics, abomasal starch infusions and the subsequent hindgut acidosis had little or no meaningful effects on biomarkers of immune activation or production variables.

Feed intake is related to changes in plasma nonesterified fatty acid concentration and hepatic acetyl CoA content following feeding in lactating dairy cows.

The relationship between hepatic acetyl CoA (AcCoA) content and dry matter intake (DMI) was evaluated using 28 multiparous Holstein cows; 14 were early postpartum (PP; 12.6 ± 3.8 d in milk) and 14 were late-lactation cows (LL; 269 ± 30 d in milk). Cows were fed once daily, and DMI was determined for the first 4h after feeding. Liver and blood samples were collected before feeding and 4h after feeding. Feed intake over the 4-h period ranged from 3.7 to 9.6 kg of dry matter and was similar for the 2 stages of lactation. Before feeding, hepatic AcCoA content was greater for PP compared with LL cows (34.4 vs. 12.5 nmol/g), and decreased over the 4h after feeding for PP only (28.7 vs. 34.4 nmol/g). The range for change in AcCoA over the 4-h period was wide for both PP (-24.3 to 10.4 nmol/g) and LL (-5.7 to 16.1 nmol/g), and was related negatively to DMI at 4h for both PP (R(2) = 0.55) and LL (R(2) = 0.31). The reduction in plasma NEFA concentration over the 4-h period was greater for PP than LL cows (-681 vs. -47 µEq/L), and was related to DMI at 4h for both PP and LL (both R(2) = 0.38). Greater DMI among cows over the first 4h after feeding might have been from a sharper reduction in supply of AcCoA in the liver for oxidation during meals because of the reduction in plasma NEFA concentration. Consistent with this is that the change in AcCoA was positively related to the reduction in plasma NEFA concentration for PP cows (R(2) = 0.31). However, change in plasma NEFA concentration was not related to change in hepatic AcCoA in LL cows, indicating that the pool of AcCoA in LL cows is not as dependent on NEFA flux to the liver as that of PP cows. Further research is required to determine production and fate of AcCoA within the timeframe of meals and the effects of feeding on energy charge in hepatic tissue.

Evaluation of propylene glycol and glycerol infusions as treatments for ketosis in dairy cows.

To evaluate propylene glycol (PG) and glycerol (G) as potential treatments for ketosis, we conducted 2 experiments lasting 4 d each in which cows received one bolus infusion per day. Blood was collected before infusion, over 240min postinfusion, as well as 24 h postinfusion. Experiment 1 used 6 ruminally cannulated cows (26±7 d in milk) randomly assigned to 300-mL infusions of PG or G (both ≥99.5% pure) in a crossover design experiment with 2 periods. Within each period, cows were assigned randomly to infusion site sequence: abomasum (A)-cranial reticulorumen (R) or the reverse, R-A. Glucose precursors were infused into the R to simulate drenching and the A to prevent metabolism by ruminal microbes. Glycerol infused in the A increased plasma glucose concentration the most (15.8mg/dL), followed by PG infused in the R (12.6mg/dL), PG infused in the A (9.11mg/dL), and G infused in the R (7.3mg/dL). Infusion of PG into the R increased plasma insulin and insulin area under the curve (AUC) the most compared with all other treatments (7.88 vs. 2.13µIU/mL and 321 vs. 31.9min×µIU/mL, respectively). Overall, PG decreased plasma BHBA concentration after infusion (-6.46 vs. -4.55mg/dL) and increased BHBA AUC (-1,055 vs. -558min ×mg/dL) compared with G. Plasma NEFA responses were not different among treatments. Experiment 2 used 8 ruminally cannulated cows (22±5 d in milk) randomly assigned to treatment sequence in a Latin square design experiment balanced for carryover effects. Treatments were 300mL of PG, 300mL of G, 600mL of G (2G), and 300mL of PG + 300mL of G (GPG), all infused into the R. Treatment contrasts compared PG with each treatment containing glycerol (G, 2G, and GPG). Propylene glycol increased plasma glucose (14.0 vs. 5.35mg/dL) and insulin (7.59 vs. 1.11µIU/mL) concentrations compared with G, but only tended to increase glucose and insulin concentrations compared with 2G. Propylene glycol increased AUC for glucose (1,444 vs. 94.3mg/dL) and insulin (326 vs. 6.58min×µIU/mL) compared with G, and tended to increase insulin AUC compared with 2G. Propylene glycol was not different from GPG for glucose, insulin, or BHBA responses. Propylene glycol decreased plasma BHBA concentration (-10.3 vs. -4.21mg/dL) and increased BHBA AUC (-1,578 vs. -1.42min ×mg/dL) compared with G, but not compared with 2G. In general, and compared with G, GPG decreased plasma NEFA concentrations after infusions and PG decreased plasma NEFA concentrations early but not late after infusions. We conclude that a 300-mL dose of PG is more effective at increasing plasma glucose concentration than G and at least as effective as 600mL of G or a combination of G and PG when administered in the cranial reticulorumen.

Saturated fat supplementation interacts with dietary forage neutral detergent fiber content during the immediate postpartum period in Holstein cows: Energy balance and metabolism.

Forty-eight multiparous cows were used in a randomized complete block design experiment with a 2×2 factorial arrangement of treatments to determine the interaction between a highly saturated free FA supplement (SFFA) and dietary forage NDF (fNDF) content on energy balance and metabolic responses in postpartum cows. Treatment diets were offered from 1 to 29 d postpartum and contained 20 or 26% fNDF and 0 or 2% SFFA (Energy Booster 100; 96.1% FA: 46.2% C18:0, and 37.0% C16:0). Overall, low fNDF versus high fNDF and 2% SFFA versus 0% SFFA increased digestible energy intake (DEI; 67.5 vs. 62.2 Mcal/d and 68.1 vs. 61.6 Mcal/d, respectively). The low fNDF diet with SFFA increased energy balance compared with the other treatments early during the treatment period, but treatment differences diminished over time. Overall, low fNDF versus high fNDF diets and 2% SFFA versus 0% SFFA improved energy balance (-13.0 vs. -16.3 Mcal/d and -12.0 vs. -17.3, respectively) decreasing efficiency of utilization of DEI for milk (milk NEL/DEI; 0.575 vs. 0.634 and 0.565 vs. 0.643). Low fNDF diets increased plasma insulin (0.308 vs. 0.137 µg/mL) and glucose concentrations (50.5 vs. 45.7mg/dL) and decreased plasma nonesterified FA (606 vs. 917µEq/L) and ß-hydroxybutyrate (9.29 vs. 16.5mg/dL) concentrations and liver triglyceride content. Compared with 0% SFFA, 2% SFFA decreased plasma nonesterified FA concentration during the first week postpartum (706 vs. 943µEq/L) and tended to decrease plasma nonesterified FA overall throughout the treatment period, but did not affect liver triglyceride content. During a glucose tolerance test, 2% SFFA increased plasma insulin concentration more in the low fNDF diet (84.5 vs. 44.6µIU/mL) than in the high fNDF diet (40.4 vs. 38.0µIU/mL). After glucose infusion, 2% SFFA increased insulin area under the curve by 64% when included in the low fNDF diet, but only by 5.2% when included in the high fNDF diet. Even though 2% SFFA did not affect weekly plasma insulin concentration, it increased plasma insulin baseline concentration before the tolerance tests. Supplementation of 2% SFFA and low fNDF diets increased DEI and improved energy balance, but decreased apparent efficiency of utilization of DEI for milk production. Fat supplementation affected energy partitioning, increasing energy balance and decreasing body condition score loss, especially in the lower fNDF diet. The decrease in body condition score loss observed was likely related to an increase in plasma insulin concentration. Feeding SFFA in a low fNDF diet during the first 29 d postpartum might have primed the cows to limit fat mobilization at the expense of milk.

Saturated fat supplementation interacts with dietary forage neutral detergent fiber content during the immediate postpartum and carryover periods in Holstein cows: Production responses and digestibility of nutrients.

Forty-eight multiparous cows were used in a randomized complete block design experiment with a 2×2 factorial arrangement of treatments to determine the interaction between a highly saturated free FA supplement (SFFA) and dietary forage neutral detergent fiber (fNDF) content on production responses and nutrient digestibility of dairy cows in the postpartum period. Treatment diets were offered from 1 to 29d postpartum (postpartum period; PP) and contained 20 or 26% fNDF (50:50 corn silage:alfalfa silage and hay, dry matter basis) and 0 or 2% SFFA [Energy Booster 100 (Milk Specialties Global, Eden Prairie, MN); 96.1% FA: 46.2% C18:0 and 37.0% C16:0]. From 30 to 71d postpartum (carryover period), a common diet (~23% fNDF, 0% SFFA) was offered to all cows to evaluate carryover effects of the treatment diets early in lactation. During the PP, higher fNDF decreased dry matter intake (DMI) by 2.0 kg/d, whereas SFFA supplementation increased it by 1.4kg/d. In addition, high fNDF with 0% SFFA decreased DMI compared with the other diets and this difference increased throughout the PP. Treatments did not affect 3.5% fat-corrected milk yield during the PP but did during the carryover period when SFFA supplementation decreased 3.5% fat-corrected milk yield for the low-fNDF diet (51.1 vs. 58.7kg/d), but not for the high-fNDF diet (58.5 vs. 58.0kg/d). During the PP, lower fNDF and SFFA supplementation decreased body condition score loss. A tendency for an interaction between fNDF and SFFA indicated that low fNDF with 2% SFFA decreased body condition score loss compared with the other diets (-0.49 vs. -0.89). During the PP, lower fNDF and 2% SFFA supplementation decreased feed efficiency (3.5% fat-corrected milk/DMI) by 0.30 and 0.23 units, respectively. The low-fNDF diet with 2% SFFA decreased feed efficiency compared with other diets early in the PP, but this difference decreased over time. Supplementation of SFFA in the PP favored energy partitioning to body reserves and limited DMI depression for the high-fNDF diet, which might allow higher-fNDF diets to be fed to cows in the PP. However, SFFA supplemented in the low-fNDF diet during the PP affected production negatively in the carryover period. Dietary fNDF and SFFA interacted, affecting performance in the PP with carryover effects when cows were fed a common diet in early lactation.

Milk production responses to dietary stearic acid vary by production level in dairy cattle.

Effects of stearic acid supplementation on feed intake and metabolic and production responses of dairy cows with a wide range of milk production (32.2 to 64.4 kg/d) were evaluated in a crossover design experiment with a covariate period. Thirty-two multiparous Holstein cows (142±55 d in milk) were assigned randomly within level of milk yield to treatment sequence. Treatments were diets supplemented (2% of diet dry matter) with stearic acid (SA; 98% C18:0) or control (soyhulls). The diets were based on corn silage and alfalfa and contained 24.5% forage neutral detergent fiber, 25.1% starch, and 17.3% crude protein. Treatment periods were 21 d with the final 4 d used for data and sample collection. Compared with the control, SA increased dry matter intake (DMI; 26.1 vs. 25.2 kg/d) and milk yield (40.2 vs. 38.5 kg/d). Stearic acid had no effect on the concentration of milk components but increased yields of fat (1.42 vs. 1.35 kg/d), protein (1.19 vs. 1.14 kg/d), and lactose (1.96 vs. 1.87 kg/d). The SA treatment increased 3.5% fat-corrected milk (3.5% FCM; 40.5 vs. 38.6 kg/d) but did not affect feed efficiency (3.5% FCM/DMI, 1.55 vs. 1.53), body weight, or body condition score compared with the control. Linear interactions between treatment and level of milk yield during the covariate period were detected for DMI and yields of milk, fat, protein, lactose, and 3.5% FCM; responses to SA were positively related to milk yield of cows. The SA treatment increased crude protein digestibility (67.4 vs. 65.5%), tended to increase neutral detergent fiber digestibility (43.6 vs. 42.3%), decreased fatty acid (FA) digestibility (56.6 vs. 76.1%), and did not affect organic matter digestibility. Fatty acid yield response, calculated as the additional FA yield secreted in milk per unit of additional FA intake, was only 13.3% for total FA and 8.2% for C18:0 plus cis-9 C18:1. Low estimated digestibility of the SA supplement was at least partly responsible for the low FA yield response. Treatment did not affect plasma insulin, glucagon, glucose, and nonesterified FA concentrations. Results show that stearic acid has the potential to increase DMI and yields of milk and milk components, without affecting conversion of feed to milk, body condition score, or body weight. Moreover, effects on DMI and yields of milk and milk components were more pronounced for higher-yielding cows than for lower-yielding cows.

Palmitic acid increased yields of milk and milk fat and nutrient digestibility across production level of lactating cows.

The effects of palmitic acid supplementation on feed intake, digestibility, and metabolic and production responses were evaluated in dairy cows with a wide range of milk production (34.5 to 66.2 kg/d) in a crossover design experiment with a covariate period. Thirty-two multiparous Holstein cows (151 ± 66 d in milk) were randomly assigned to treatment sequence within level of milk production. Treatments were diets supplemented (2% of diet DM) with palmitic acid (PA; 99% C16:0) or control (SH; soyhulls). Treatment periods were 21 d, with the final 4 d used for data and sample collection. Immediately before the first treatment period, cows were fed the control diet for 21 d and baseline values were obtained for all variables (covariate period). Milk production measured during the covariate period (preliminary milk yield) was used as covariate. In general, no interactions were detected between treatment and preliminary milk yield for the response variables measured. The PA treatment increased milk fat percentage (3.40 vs. 3.29%) and yields of milk (46.0 vs. 44.9 kg/d), milk fat (1.53 vs. 1.45 kg/d), and 3.5% fat-corrected milk (44.6 vs. 42.9 kg/d), compared with SH. Concentrations and yields of protein and lactose were not affected by treatment. The PA treatment did not affect dry matter (DM) intake or body weight, tended to decrease body condition score (2.93 vs. 2.99), and increased feed efficiency (3.5% fat-corrected milk/DM intake; 1.60 vs. 1.54), compared with SH. The PA treatment increased total-tract digestibility of neutral detergent fiber (39.0 vs.35.7%) and organic matter (67.9 vs. 66.2%), but decreased fatty acid (FA) digestibility (61.2 vs. 71.3%). As total FA intake increased, total FA digestibility decreased (R(2) = 0.51) and total FA absorbed increased (quadratic R(2) = 0.82). Fatty acid yield response, calculated as the additional FA yield secreted in milk per unit of additional FA intake, was 11.7% for total FA and 16.5% for C16:0 plus cis-9 C16:1 FA. The PA treatment increased plasma concentration of nonesterified FA (101 vs. 90.0 µEq/L) and cholecystokinin (19.7 vs. 17.6 pmol/L), and tended to increase plasma concentration of insulin (10.7 vs. 9.57 µ IU/mL). Results show that palmitic acid fed at 2% of diet DM has the potential to increase yields of milk and milk fat, independent of production level without increasing body condition score or body weight. However, a small percentage of the supplemented FA was partitioned to milk.

Level of nutrient intake affects mammary gland gene expression profiles in preweaned Holstein heifers.

Bovine mammary parenchyma (PAR) and fat pad (MFP) development are responsive to preweaning level of nutrient intake. We studied transcriptome alterations in PAR and MFP from Holstein heifer calves (n=6/treatment) fed different nutrient intakes from birth to ca. 65 d age. Conventional nutrient intake received 441 g of dry matter (DM)/d of a control milk replacer (MR) [CON; 20% crude protein (CP), 20% fat, DM basis]. Calves in the accelerated nutrition groups received 951 g/d of high-protein/low-fat MR (HPLF; 28% CP, 20% fat, DM basis), 951 g/d of high-protein/high-fat MR (HPHF; 28% CP, 28% fat, DM basis), or 1,431 g/d of HPHF (HPHF+) MR. Out of 13,000 genes evaluated, over 1,500 differentially expressed genes (DEG) were affected (false discovery rate <0.10) by level of nutrient intake in PAR or MFP. Feeding HPLF versus CON resulted in the most dramatic changes in gene expression, with 278 and 588 DEG having ≥1.5-fold change in PAR and MFP. In PAR, the most-altered molecular functions were associated with metabolism of the cell (molecular transport and lipid metabolism) with most of the genes downregulated in HPLF versus CON. In MFP, DEG also were primarily associated with metabolism but changes also occurred in genes linked to cell morphology, cell-to-cell signaling, and immune response. Compared with CON, feeding HPHF or HPHF+ did not result in substantial additional effects on DEG beyond those observed with HPLF. The pentose phosphate, mitochondrial dysfunction, and ubiquinone biosynthesis pathways were among the most enriched due to HPLF versus CON in PAR and were inhibited, whereas glycosphingolipid biosynthesis, arachidonic acid metabolism, and eicosanoid synthesis pathways were among the most enriched due to HPLF versus CON in MFP and were inhibited. These responses suggest that, in PAR, doubling nutrient intake from standard feeding rates inhibited energy metabolism and activity of oxidative pathways that partly serve to protect cells against oxidative stress. The MFP in those heifers appeared to decrease production of lipid-derived metabolites that may play roles in signaling pathways within the adipocyte. Overall, results indicated that prepubertal/preweaned mammary transcriptome is responsive to long-term enhanced nutrient supply to achieve greater growth rates before weaning. The biological significance of these results to future milk production remains to be elucidated.

Adipose tissue lipogenic gene networks due to lipid feeding and milk fat depression in lactating cows.

Dietary lipid supplements have been extensively evaluated for their effects on mammary tissue mRNA abundance, including the classical lipogenic genes ACACA, SCD, FASN, and the transcription regulators SREBF1, THRSP, and PPARG. Novel gene isoforms with key regulatory roles in triacylglycerol synthesis have been recently identified including LPIN1 and AGPAT6. Transcriptional networks (i.e., genes whose mRNA expression is regulated by a transcription factor or nuclear receptor) coordinate adipogenesis and lipid filling in nonruminant adipose tissue. To investigate whether long-term milk fat depression affects adipogenic networks in subcutaneous adipose tissue, we characterized mRNA expression via quantitative PCR of 20 genes in cows fed saturated and polyunsaturated lipid for 3 wk. Adipose tissue from cows fed a control diet, control with fish (10 g/kg of dry matter) and soybean oil (25 g/kg of dry matter) (FSO), or control with saturated lipid (35 g/kg, EB100; Energy Booster 100, Milk Specialties, Dundee, IL) was biopsied after 21 d of feeding. Milk production did not differ across treatments (averaged 32 kg +/- 2.8 kg/d during the 21 d) but dry matter intake (DMI) decreased in cows fed FSO versus controls (averaged 18 vs. 22 kg/d during the 21 d). Despite the decrease in DMI, FSO resulted in similar energy intake as EB100 during the last 2 wk of the study. Cows fed FSO had a gradual decline in milk fat and energy yield leading to an overall 25% decrease in milk fat yield during the study (averaged 0.90 vs. 1.2 kg/d) compared with control or EB100. Thus, during the 21-d study, FSO led to a gradual increase in intake energy available for adipose tissue deposition. Relative mRNA expression of LPL and SCD as well as ADFP (coding for a protein involved in lipid droplet formation) and LPIN1 (coding for a protein involved in diacylglycerol synthesis/transcriptional regulation) was upregulated with FSO relative to other diets. Expression of the transcription regulator THRSP tended to be greater in cows fed FSO. Overall, results suggest that long-term milk fat depression caused by feeding FSO provided additional energy as well as long-chain fatty acids that, coupled with upregulation of a subset of adipogenic genes in subcutaneous adipose tissue, might have resulted in greater tissue lipid deposition.

Regeneration of rat calvarial defects using a bioabsorbable membrane technique: influence of collagen cross-linking.

The aim of the present study was to examine the influence of cross-linking on collagen membranes used for guided bone regeneration of calvarial defects in rats. In 48 Wistar rats, divided equally into 4 groups, 1 control and 3 experimental, standardized transosseous circular calvarial defects were made midparietally. In the control group, the defect was only covered by the soft tissue flap while in the 3 experimental groups, 3 differently cross-linked collagen membranes were interposed between the osseous defect and the overlying flap before suturing. The healing was assessed at 10, 20, and 30 days after surgery. The results showed that augmenting the degree of collagen cross-linking diminished the membrane resorption rate. Compared to the sham-operated sites, the membrane protected defects showed significantly more bone regeneration (on average 4 times more) as attested by histology and measured by histomorphometric analysis. Although the bone gain seemed to augment with increasing degrees of cross-linking, the results within the 3 experimental groups were not statistically different. Since longer healing periods might have been necessary to substantiate results within experimental groups, a study is currently undertaken to evaluate this aspect. This study demonstrated the efficacy of collagen membranes in guiding bone regeneration, as well as the importance of the type and degree of cross-linking.