Association of residual feed intake with abundance of ruminal bacteria and biopolymer hydrolyzing enzyme activities during the peripartal period and early lactation in Holstein dairy cows

Background: Residual feed intake(RFI) in dairy cattle typically calculated at peak lactation is a measure of feed efficiency independent of milk production level. The objective of this study was to evaluate differences in ruminal bacteria, biopolymer hydrolyzing enzyme activities, and overall performance between the most-and the leastefficient dairy cows during the peripartal period. Twenty multiparous Holstein dairy cows with daily ad libitum access to a total mixed ration from d-10 to d 60 relative to the calving date were used. Cows were classified into most-efficient(i.e. with low RFI, n = 10) and least-efficient(i.e. with high RFI, n = 10) based on a linear regression model involving dry matter intake(DMI), fat-corrected milk(FCM), changes in body weight(BW), and metabolic BW.Results: The most-efficient cows had ~ 2.6 kg/d lower DMI at wk 4, 6, 7, and 8 compared with the least-efficient cows. In addition, the most-efficient cows had greater relative abundance of total ruminal bacterial community during the peripartal period. Compared with the least-efficient cows, the most-efficient cows had 4-fold greater relative abundance of Succinivibrio dextrinosolvens at d-10 and d 10 around parturition and tended to have greater abundance of Fibrobacter succinogenes and Megaspheara elsdenii. In contrast, the relative abundance of Butyrivibrio proteoclasticus and Streptococcus bovis was lower and Succinimonas amylolytica and Prevotella bryantii tended to be lower in the most-efficient cows around calving. During the peripartal period, the most-efficient cows had lower enzymatic activities of cellulase, amylase, and protease compared with the least-efficient cows.Conclusions: The results suggest that shifts in ruminal bacteria and digestive enzyme activities during the peripartal period could, at least in part, be part of the mechanism associated with better feed efficiency in dairy cows.

Rumen-protected methionine during the peripartal period in dairy cows and its effects on abundance of major species of ruminal bacteria

Background: Extensive degradation of amino acids in the rumen via microbial deamination decreases the postruminal availability of dietary indispensable amino acids. Together with the normal decrease in voluntary dry matter intake(DMI) around parturition in dairy cows, microbial metabolism contributes to a markedly negative balance of indispensable amino acids, including methionine which may be the first-limiting for milk production. The main objective of the current study was to profile changes in major bacterial species with key functions in cel ulose and hemicel ulose digestion, xylan breakdown, proteolytic action, propionic acid production, lactate utilization and ruminal biohydrogenation in cows supplemented with rumen-protected Methionine(SM; Smartamine M, Adisseo NA, Alpharetta,GA, USA) from-23 through 30 d relative to parturition. Because ~90% of the methionine in SM bypasses the rumen,~10% of the methionine is released into the rumen and can be utilized by microbes.Results: As expected, there was an increase in overall DMI after parturition(Day, P < 0.05) during which cows consumed on average 19.6 kg/d versus 13.9 kg/d in the prepartum period. The postpartum diet contained greater concentrations of lipid and highly-fermentable carbohydrate from corn grain, which likely explains the increases in the relative abundance of Anaerovibrio lipolytica, Megasphaera elsdenii, Prevotella bryantii, Selenomonas ruminantium,Streptococcus bovis, and Succinimonas amylolytica. Despite similar DMI prepartum, cows fed SM had greater(Treatment × Day, P < 0.05) abundance prepartum of Fibrobacter succinogenes, Succinimonas amylolytica, and Succinivibrio dextrinosolvens. However, the greater DMI in cows fed SM after parturition(19.6 kg/d versus 13.9 kg/d) was associated with lower abundance of Fibrobacter succinogenes(2.13 × 10-3 versus 2.25 × 10-4) and Selenomonas ruminantium(2.98 × 10-1 versus 4.10 × 10-1). A lower abundance(Day, P < 0.05) was detected on d 20 compared with d-10 for Fibrobacter succinogenes and Succinivibrio dextrinosolvens. The relative abundance of Butyrivibrio proteoclasticus and Eubacterium ruminantium was stable across treatment and time.(Continued from previous page)Conclusions: In diets with proper balance of rumen-degradable protein and fermentable carbohydrate, the smal fraction of Methionine released from the rumen-protected supplement did not seem to compromise growth of major bacterial species in the rumen. In fact, it had a positive effect on 3 major species prepartum when DMI was similar between groups. Because the actual requirements of Methionine(and Lysine, for example) by the cow during the transition period are unknown, it appears warranted to study the rumen microbiome as it relates to supply of rumen-protected amino acids.

Structures and characteristics of carbohydrates in diets fed to pigs: a review

The current paper reviews the content and variation of fiber fractions in feed ingredients commonly used in swine diets.Carbohydrates serve as the main source of energy in diets fed to pigs.Carbohydrates may be classified according to their degree of polymerization: monosaccharides,disaccharides,oligosaccharides,and polysaccharides.Digestible carbohydrates include sugars,digestible starch,and glycogen that may be digested by enzymes secreted in the gastrointestinal tract of the pig.Non-digestible carbohydrates,also known as fiber,may be fermented by microbial populations along the gastrointestinal tract to synthesize short-chain fatty acids that may be absorbed and metabolized by the pig.These non-digestible carbohydrates include two disaccharides,oligosaccharides,resistant starch,and non-starch polysaccharides.The concentration and structure of non-digestible carbohydrates in diets fed to pigs depend on the type of feed ingredients that are included in the mixed diet.Cellulose,arabinoxylans,and mixed linked β-(1,3)(1,4)-D-glucans are the main cell wall polysaccharides in cereal grains,but vary in proportion and structure depending on the grain and tissue within the grain.Cell walls of oilseeds,oilseed meals,and pulse crops contain cellulose,pectic polysaccharides,lignin,and xyloglucans.Pulse crops and legumes also contain significant quantities of galacto-oligosaccharides including raffinose,stachyose,and verbascose.Overall,understanding the structure,characteristics and measurable chemical properties of fiber in feed ingredients may result in more accurate diet formulations,resulting in an improvement in the utilization of energy from less expensive high-fiber ingredients and a reduction in reliance on energy from more costly cereal grains.

Multifaceted role of one-carbon metabolism on immunometabolic control and growth during pregnancy, lactation and the neonatal period in dairy cattle

Dairy cattle undergo dramatic metabolic, endocrine, physiologic and immune changes during the peripartal period largely due to combined increases in energy requirements for fetal growth and development, milk production, and decreased dry matter intake. The negative nutrient balance that develops results in body fat mobilization,subsequently leading to triacylglycerol(TAG) accumulation in the liver along with reductions in liver function,immune dysfunction and a state of inflammation and oxidative stress. Mobilization of muscle and gluconeogenesis are also enhanced, while intake of vitamins and minerals is decreased, contributing to metabolic and immune dysfunction and oxidative stress. Enhancing post-ruminal supply of methyl donors is one approach that may improve immunometabolism and production synergistically in peripartal cows. At the cellular level, methyl donors(e.g. methionine, choline, betaine and folic acid) interact through one-carbon metabolism to modulate metabolism,immune responses and epigenetic events. By modulating those pathways, methyl donors may help increase the export of very low-density lipoproteins to reduce liver TAG and contribute to antioxidant synthesis to alleviate oxidative stress. Thus, altering one-carbon metabolism through methyl donor supplementation is a viable option to modulate immunometabolism during the peripartal period. This review explores available data on the regulation of one-carbon metabolism pathways in dairy cows in the context of enzyme regulation, cellular sensors and signaling mechanisms that might respond to increased dietary supply of specific methyl donors. Effects of methyl donors beyond the one-carbon metabolism pathways, including production performance, immune cell function,mechanistic target or rapamycin signaling, and fatty acid oxidation will also be highlighted. Furthermore, the effects of body condition and feeding system(total mixed ration vs. pasture) on one-carbon metabolism pathways are explored. Potential effects of methyl donor supply during the pepartum period on dairy calf growth and development also are discussed. Lastly, practical nutritional recommendations related to methyl donor metabolism during the peripartal period are presented. Nutritional management during the peripartal period is a fertile area of research, hence, underscoring the importance for developing a systems understanding of the potential immunometabolic role that dietary methyl donors play during this period to promote health and performance.

Maternal supply of methionine during late-pregnancy enhances rate of Holstein calf development in utero and postnatal growth to a greater extent than colostrum source

Background: Pregnancy and early life are critical periods of plasticity during which the fetus and neonate may be influenced by environmental factors such as nutrition.Maternal methionine(Met) supply in non-ruminants during pregnancy can affect offspring development and growth.Thus,the objective of this study was to investigate if increasing Met supply during late-pregnancy affects developmental parameters of the calf at birth and if either maternal Met or colostrum from Met-fed cows alters calf growth.Calves born to Holstein cows individually-fed a basal control [CON; 1.47 Mcal/kg dry matter(DM) and 15.3% crude protein] diet with no added Met or CON plus ethylcellulose rumen-protected Met(MET; Mepron? at 0.09% of diet DM; Evonik Nutrition & Care GmbH,Germany)during the last 28 ± 2 d of pregnancy were used.A total of 39 calves were in CON(n = 22 bulls,17 heifers) and 42 in MET(n = 20 bulls,22 heifers).At birth,calves were randomly allocated considering dam treatment and colostrum as fol ows: 1) calves from CON cows and colostrum from CON cows(n = 21); 2) calves from CON cows and colostrum from MET cows(n = 18); 3) calves from MET cows and colostrum from MET cows(n = 22); and 4) calves from MET cows and colostrum from CON cows(n = 20).Al calves were housed,managed,and fed individual y during the first 9 wk of life.Results: Despite greater daily DM intake pre-partum in cows fed MET(15.7 vs.14.4 ± 0.12 kg/d,P < 0.05),colostrum quality and quantity were not affected by maternal diet.At birth,MET calves had greater(P ≤ 0.05) body weight(BW,44.1 vs.42.1 ± 0.70 kg),hip height(HH,81.3 vs.79.6 ± 0.53 cm) and wither height(WH,77.8 vs.75.9 ± 0.47 cm).In contrast,concentrations of His,Lys,and Asn in plasma were lower(P ≤ 0.05) in MET calves.Regardless of colostrum source,the greater BW,HH,and WH in MET calves at birth persisted through 9 wk of age resulting in average responses of + 3.1 kg BW,+ 1.9 cm HH,and + 1.8 cm WH compared with CON.Average daily gain during the 9 wk was(P < 0.05) 0.72 ± 0.02 kg/d in MET compared with 0.67 ± 0.02 kg/d in CON calves.Respiratory scores were normal and did not differ(P > 0.05) due to maternal Met supply or colostrum source.However,fecal scores tended to be lower(P ≤ 0.10) in MET calves regardless of colostrum source.Conclusions: Increasing the maternal supply of MET during late-pregnancy enhanced growth in utero as wel as during the pre-weaning and early post-weaning periods.Although the ～ 1 kg/d greater DM intake during the last 2–3 wk prior to parturition could explain a portion of the 2 kg extra body mass of MET calves at birth,other mechanisms potential y encompassing nutrient assimilation efficiency likely played a role.Assessing the exact mechanisms sensitive to supply of Met or total amino acid supply during the latter stages of growth in utero merit further research.

Influence of the concentration of dietary digestible calcium on growth performance,bone mineralization, plasma calcium, and abundance of genes involved in intestinal absorption of calcium in pigs from 11 to 22 kg fed diets with different concentrations of

Background: A 21-day experiment was conducted to test the hypothesis that Ca requirements to maximize growth performance expressed as the standardized total tract digestible(STTD) Ca to STTD P ratio is less than 1.40:1. The second hypothesis was that increasing dietary Ca increases plasma Ca concentration and downregulates abundance of genes related to Ca absorption(TRPV6, S100 G, and ATP2 B1) in the duodenum, and tight junction proteins(OCLN, CLDN1, and ZO1) in the duodenum and ileum.Methods: Twenty corn-soybean meal diets were formulated using a 4 × 5 factorial design with diets containing 0.16%, 0.33%, 0.42%, or 0.50% STTD P, and 0.14%, 0.29%, 0.44%, 0.59%, or 0.74% STTD Ca. Six hundred and forty pigs(initial weight: 11.1 ± 1.4 kg) were allotted to 20 diets and 5 blocks in a randomized complete block design. On day21, weights of pigs and feed left in feeders were recorded and blood, duodenal tissue, ileal mucosa, and the right femur were collected from 1 pig per pen. Abundance of m RNA was determined in duodenal and ileal tissue via quantitative RT-PCR. Data were analyzed using a response surface model.Results: The predicted maximum ADG(614 g), G:F(0.65), and bone ash(11.68 g) was obtained at STTD Ca:STTD P ratios of 1.39:1, 1.25:1, and 1.66:1, respectively, when STTD P was provided at the requirement(0.33%). If dietary STTD P was below the requirement, increasing dietary Ca resulted in reduced(P < 0.05) ADG and G:F. However, if dietary STTD P was above the requirement, negative effects(P < 0.05) on ADG and G:F of increasing STTD Ca were observed only if dietary STTD Ca exceeded 0.6%. Plasma Ca concentration was positively affected by STTD Ca over the range studied(quadratic, P < 0.01) and negatively affected by increasing STTD P(linear, P < 0.01). There was a linear negative effect(P < 0.05) of STTD Ca on the abundance of S100 G, TRPV6, OCLN, and ZO1 in duodenum, and CLDN and ZO1 in ileum.Conclusions: The STTD Ca:STTD P ratio needed to maximize growth performance of 11-to 25-kg pigs is less than1.40:1, if P is at the estimated requirement. Increasing dietary Ca reduces transcellular absorption of Ca and increases paracellular absorption of Ca.

Body condition score prior to parturition is associated with plasma and adipose tissue biomarkers of lipid metabolism and inflammation in Holstein cows

Background: Previous research has revealed a strong inflammatory response within adipose(AT) tissue during the transition into lactation. Whether this effect is a result of oxidative stress induced by lipolysis and fatty acid oxidation associated with differences in prepartum body condition score remains to be determined. The objectives of this study were to investigate systemic biomarkers of energy balance and inflammation and the expression of lipid metabolismand inflammation-related genes in AT during the transition period in dairy cows.Results: Twenty multiparous Holstein cows were retrospectively divided by body condition score(BCS) prior to parturition into two groups(10 cows/group): BCS ≤ 3.25(Lo BCS) and BCS ≥ 3.75(Hi BCS). Subcutaneous adipose tissue was biopsied from the tail-head region at d-10, 7 and 20 relative to parturition. Plasma was used to evaluate biomarkers of energy balance(EBAL) [free fatty acids(NEFA), glycerol, insulin] and inflammation [IL-1β, haptoglobin, myeloperoxidase, and reactive oxygen metabolites(ROM)]. Although insulin concentration was not affected by BCS, NEFA was overal greater and glycerol lower in HiB CS cows. Greater activity of myeloperoxidase in plasma coincided with increased haptoglobin and IL-1β postpartum in LoB CS cows.Among genes related with oxidative stress, the expression of the cytosolic antioxidant enzyme SOD1 was greater in LoB CS compared to HiB CS. Cows in LoB CS compared with HiB CS had greater overal expression of ABDH5 and ATGL along with ADIPOQ, indicating enhanced basal lipolysis and secretion of adiponectin. Expression of CPT1 A, ACADVL, and ACOX1 was greater overal in HiB CS than LoB CS indicating enhanced NEFA oxidation. Although the temporal increase in plasma NEFA regardless of BCS coincided with the profile of CPT1 A, the gradual decrease in genes related with reesterification of NEFA(PCK1) and glycerol efflux(AQP7) coupled with an increase in glycerol kinase(GK) suggested some stimulation of NEFA utilization within adipose tissue. This idea is supported in part by the gradual decrease in insulin regardless of BCS. Although expression of the inflammation-related gene toll-like receptor 4( TLR4) was greater in Hi BCS versus Lo BCS cows at-10 d, expression of TLR9 was greater in Hi BCS versus Lo BCS at 20 d.These profiles did not seem to be associated with concentrations of pro-inflammatory biomarkers or ROM.Conclusions: Overall, data indicated that cows with BCS 3.25 or lower before calving experienced greater alterations in systemic inflammation and basal lipolysis without excessive increases in NEFA plasma concentrations.Despite the greater plasma NEFA around parturition, cows with BCS 3.75 or higher seemed to have a more active system for catabolism of NEFA and utilization of glycerol within adipose tissue. A linkage between those pathways and risk of disorders postpartum remains to be determined.

Knockout of butyrophilin subfamily 1 member A1(BTN1A1) alters lipid droplet formation and phospholipid composition in bovine mammary epithelial cells

Background: Milk lipids originate from cytoplasmic lipid droplets(LD) that are synthesized and secreted from mammary epithelial cells by a unique membrane-envelopment process. Butyrophilin 1 A1(BTN1 A1) is one of the membrane proteins that surrounds LD, but its role in bovine mammary lipid droplet synthesis and secretion is not well known.Methods: The objective was to knockout BTN1 A1 in bovine mammary epithelial cells(BMEC) via the CRISPR/Cas9 system and evaluate LD formation, abundance of lipogenic enzymes, and content of cell membrane phospholipid(PL) species. Average LD diameter was determined via Oil Red O staining, and profiling of cell membrane phospholipid species via liquid chromatography-tandem mass spectrometry(LC-MS/MS).Results: Lentivirus-mediated infection of the Cas9/sg RNA expression vector into BMEC resulted in production of a homozygous clone BTN1 A1^((-/-)). The LD size and content decreased following BTN1 A1 gene knockout. The m RNA abundance of fatty acid synthase(FASN) and peroxisome proliferator-activated receptor-gamma(PPARG) was downregulated in the BTN1 A1^((-/-))clone. Subcellular analyses indicated that BTN1 A1 and LD were co-localized in the cytoplasm. BTN1 A1 gene knockout increased the percentage of phosphatidylethanolamine(PE) and decreased phosphatidylcholine(PC), which resulted in a lower PC/PE ratio.Conclusions: Results suggest that BTN1 A1 plays an important role in regulating LD synthesis via a mechanism involving membrane phospholipid composition.

Hepatic metabolomics and transcriptomics to study susceptibility to ketosis in response to prepartal nutritional management

Background:Ketosis in dairy cows is associated with body fat mobilization during the peripartal period.Sub-clinical and clinical ketosis arise more frequently in cows that are overfed energy during the entire dry(last 50 to 45 days prior to parturition)or close-up period(last^28 days prepartum).Methods:A retrospective analysis was performed on 12 cows from a larger cohort that were fed a higher-energy diet[1.54 Mcal/kg of dry matter(DM);35.9%of DM corn silage and 13%of DM ground corn]during the close-up dry period,of which 6 did not develop clinical ketosis(OVE,0.83 mmol/L plasma hydroxybutyrate;BHB)and 6 were diagnosed with clinical ketosis(KET,1.4 mmol/L BHB)during the first week postpartum.A whole-transcriptome bovine microarray(Agilent Technologies)and metabolomics(GC-MS,LC-MS;Metabolon~?Inc.)were used to perform transcript and metabolite profiling of liver tissue harvested at-10 days relative to parturition which allowed to establish potential associations between prepartal transcriptome/metabolome profiles and susceptibility to clinical ketosis postpartum.Results:Cows in KET had greater(P=0.01)overall body weight between-2 and 1 week around parturition,but similar body condition score than OVE.Although dry matter intake(DMI)did not differ prepartum,KET cows had lower(P<0.01)DMI and similar milk yield as OVE cows during the first week postpartum.Transcriptome analysis revealed a total of 3065 differentially expressed genes(DEG;P≤0.05)in KET.Metabolomics identified 15 out of 313 total biochemical compounds significantly affected(P≤0.10)in KET.Among those,greater concentrations(P≤0.06,+2.3-fold)of glycochenodeoxycholate in KET cows also have been detected in humans developing non-alcoholic fatty liver disease.Bioinformatics analysis using the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway database and the DEG revealed that,among the top 20 most-impacted metabolic pathway categories in KET,65%were overall downregulated.Those included’Metabolism of cofactors and vitamins’,’Biosynthesis of other secondary metabolites’,’Lipid’,’Carbohydrate’,and’Glycan biosynthesis and metabolism’.The lower relative concentration of glucose-6-phosphate and marked downregulation of fructose-1,6-bisphosphatase 2 and pyruvate dehydrogenase kinase 4 support a strong impairment in gluconeogenesis in prepartal liver of cows developing KET postpartum.Among the top 20 most-impacted non-metabolic pathways,85%were downregulated.Pathways such as’mTOR signalling’and’Insulin signalling’were among those.’Ribosome’,’Nucleotide excision repair’,and’Adherens junctions’were the only upregulated pathways in cows with KET.Conclusions:The combined data analyses revealed more extensive alterations of the prepartal liver transcriptome than metabolome in cows overfed energy and developing ketosis postpartum.The causative link between these tissue-level adaptations and onset of clinical ketosis needs to be studied further.

Individual and Combined Effects of Nucleotides and Human Milk Oligosaccharides on Proliferation, Apoptosis and Necrosis in a Human Fetal Intestinal Cell Line

Nucleotides (NT) and human milk oligosaccharides (HMO) individually affect epithelial cell growth, but their combined effects had not been studied. Herein, the impact of NT and HMO on cell proliferation, apoptosis, necrosis and cell cycle in the fetal epithelial cell line (FHs-74 Int) was determined. Cells were incubated with media containing 2.5% FBS and no epidermal growth factor (Control);fucosyllactose (FL) mix [85% 2’FL/15% 3’FL], sialyllactose (SL) mix [40% 6’SL/10% 3’SL/50% sialic acid (SA)] or LNnT at 125, 250, 500 or 1000 μg/mL with and without 250 μg/mL NT (43% CMP, 18.5% UMP, 16.4% AMP, and 22.0% GMP) for 24 or 72 h. NT alone significantly increased proliferation, but did not affect cell cycle or apoptosis/necrosis. All HMO treatments at 1000 μg/mL significantly decreased proliferation and some were also inhibitory at 250 or 500 μg/mL. When NT and HMO were simultaneously added, NT ameliorated the anti-proliferative effect of HMO. FL significantly increased cells in S phase and SL and LNnT treatments significantly increased cells in G2/M and S phases, which concomitantly decreased cells in G0/G1. HMO with NT significantly decreased the percent of cells in the G2/M phase compared to HMO alone. Higher HMO doses significantly increased the percentage of apoptotic and necrotic cells compared to control. In conclusion, HMO reduced cell proliferation and this effect is partially ameliorated by NT. It appears that HMO initially induced apoptosis/necrosis, which was later evidenced by G2/M cell cycle arrest and decreased proliferation.

Prepartum dietary energy intake alters adipose tissue transcriptome profiles during the periparturient period in Holstein dairy cows

Background:The aim of the study was to investigate the effect of energy overfeeding during the dry period on adipose tissue transcriptome profiles during the periparturient period in dairy cows.Methods:Fourteen primiparous Holstein cows from a larger cohort receiving a higher-energy diet(1.62 Mcal of net energy for lactation/kg of dry matter;15%crude protein)for ad libitum intake to supply 150%(OVR)or 100%(CTR)of energy requirements from dry off until parturition were used.After calving,all cows received the same lactation diet.Subcutaneous adipose tissue(SAT)biopsies were collected at−14,1,and 14d fromparturition(d)and used for transcriptome profiling using a bovine oligonucleotide microarray.Data mining of differentially expressed genes(DEG)between treatments and due to sampling time was performed using the Dynamic Impact Approach(DIA)and Ingenuity Pathway Analysis(IPA).Results:There was a strong effect of over-feeding energy on DEG with 2434(False discovery rate-corrected P<0.05)between OVR and CTR at−14 d,and only 340 and 538 at 1 and 14 d.The most-impacted and activated pathways in the Kyoto Encyclopedia of Genes and Genomes(KEGG)database that were highlighted by DIA analysis at−14 d in OVR vs.CTR included 9 associated with carbohydrate metabolism,with‘Pyruvate metabolism’,‘Glycolysis/gluconeogenesis’,and‘Pentose phosphate pathway’among the most-activated.Not surprisingly,OVR led to marked activation of lipid metabolism(e.g.‘Fatty acid biosynthesis’and‘Glycerolipid metabolism’).Unexpected metabolic pathways that were activated at−14 d in OVR included several related to metabolism of amino acids(e.g.branched chain)and of cofactors and vitamins(thiamin).Among endocrine and immune system pathways,at−14 d OVR led to marked activation of‘PPAR signalling’and‘Antigen processing and presentation’.Among key pathways affected over time in OVR,a number were related to translation(e.g.mTOR signaling),endocrine/immune signaling(CXCR4 and IGF1),and lipid metabolism(oxidative phosphorylation)with greater activation in OVR vs.CTR specifically at−14 d.Although statistical differences for several pathways in OVR vs.CTR nearly disappeared at 1 and 14 vs.−14 d,despite the well-known catabolic state of adipose depots after calving,the bioinformatics analyses suggested important roles for a number of signaling mechanisms at−14 vs.14 than 1 vs.-14 d.This was particularly evident in cows fed to meet predicted energy requirements during the dry period(CTR).Conclusions:Data underscored a strong activation by overfeeding energy of anabolic processes in the SAT exclusively prepartum.The study confirmed that higher-energy diets prepartum drive a transcriptional cascade of events orchestrated in part by the activation of PPARγthat regulate preadipocyte differentiation and lipid storage in SAT.Novel aspects of SAT biology to energy overfeeding or change in physiologic state also were uncovered,including the role of amino acid metabolism,mTOR signaling,and the immune system.

Effect of fiber supplementation on the microbiota in critically ill patients

AIM:To determine tolerance to fiber supplementation of semi-elemental tube feeds in critically ill patients and measure its effect on colonic microbiota and fermentation.METHODS:Thirteen intensive care unit patients receiving jejunal feeding with a semi-elemental diet for predominantly necrotizing pancreatitis were studied.The study was divided into 2 parts:first,short-term (3-9 d)clinical tolerance and colonic fermentation as assessed by fecal short chain fatty acid(SCFA)concentrations and breath hydrogen and methane was measured in response to progressive fiber supplementation increasing from 4 g tid up to normal requirement levels of 8 g tid;second,4 patients with diarrhea were studied for 2-5 wk with maximal supplementation to additionally assess its influence on fecal microbiota quantitated by quantitative polymerase chain reaction (qPCR)of microbial 16S rRNA genes and Human Intestinal Tract Chip(HITChip)microarray analysis.Nearly all patients were receiving antibiotics(10/13)and acid suppressants(11/13)at some stage during the studies.RESULTS:In group 1,tolerance to progressive fiber supplementation was good with breath hydrogen and methane evidence(P=0.008 and P<0.0001,respectively)of increased fermentation with no exacerbation of abdominal symptoms and resolution of diarrhea in 2 of 4 patients.In group 2 before supplementation,fecal microbiota mass and their metabolites,SCFA,were dramatically lower in patients compared to healthy volunteers.From qPCR and HITChip analyses we calculated that there was a 97%reduction in the predominant potential butyrate producers and starch degraders.Following 2-5 wk of fiber supplementation there was a significant increase in fecal SCFA(acetate 28.4±4.1μmol/g to 42.5±3.1μmol/g dry weight,P=0.01;propionate 1.6±0.5 vs 6.22±1.1,P=0.006 and butyrate 2.5±0.6 vs 5.9±1.1,P=0.04)and microbial counts of specific butyrate producers,with resolution of diarrhea in 3 of 4 patients.CONCLUSION:Conventional management of critically ill patients,which includes the use of elemental diets and broad-spectrum antibiotics,was associated with gross suppression of the colonic microbiota and their production of essential colonic fuels,i.e.,SCFA.Our investigations show that fiber supplementation of the feeds has the potential to improve microbiota mass and function,thereby reducing the risks of diarrhea due to dysbiosis.

Digestibility and metabolism of copper in diets for pigs and influence of dietary copper on growth performance,intestinal health,and overall immune status:a review

The current contribution reviews absorption and metabolism of copper(Cu),Cu deficiency,Cu toxicity,Cu bioavailability,and effects of pharmacological levels of Cu on growth performance and intestinal health of pigs.Copper is a micro mineral involved in metabolic reactions including cellular respiration,tissue pigmentation,hemoglobin formation,and connective tissue development.Copper is mostly absorbed in the upper gastrointestinal tract,particularly in the duodenum,but some Cu is absorbed in the stomach.One way to evaluate the efficacy of sources of Cu is to measure relative bioavailability where responses include tissue concentrations of Cu,concentrations of metalloproteins,and enzymatic activity of animals fed test diets containing graded levels of Cu.The requirement for Cu by pigs is 5 to 10 mg/kg diet,however,Cu can be included at growth-promoting levels(i.e.,75 to 250 mg/kg diet)in diets for weanling and growing pigs to reduce post-weaning diarrhea and improve growth performance.The consistently observed improvement in growth performance upon Cu supplementation is likely a result of increases in lipase activity,growth hormone secretion,and expression of genes involved in postabsorptive metabolism of lipids.The growth-promoting effects of dietary Cu have also been attributed to its bacteriostatic and bactericidal properties because Cu may change bacterial populations in the intestine,and thereby reduce inflammation caused by pathogens.However,further research is needed to determine potential interactions between Cu and non-nutritive feed additives(e.g.,enzymes,probiotics,phytobiotics),and the optimum quantity of Cu as well as the optimum duration of feeding supplemental Cu in diets for pigs should be further investigated.These gaps needs to be addressed to maximize inclusion of Cu in diets to improve growth performance while minimizing diseases and mortality.

Peripheral leukocyte and endometrium molecular biomarkers of inflammation and oxidative stress are altered in peripartal dairy cows supplemented with Zn,Mn,and Cu from amino acid complexes and Co from Co glucoheptonate

Background: Immune dysfunction and a higher risk of uterine infections are characteristics of the transition into lactation in dairy cows. The supply of complexed trace minerals, which are more bioavailable, could help overcome the greater needs of these nutrients in tissues around parturition and early lactation.Results: Twenty Holstein cows received an oral bolus with a mix of inorganic trace minerals(INO) or complexed trace minerals(AAC) to achieve 75, 65, 11, and 1 ppm supplemental Zn, Mn, Cu, and Co, respectively, in the total diet dry matter from -30 d through +30 d relative to parturition. Blood for polymorphonuclear leukocyte(PMNL) isolation was collected at-30,-15, +10, and + 30 d relative to parturition, whereas endometrium biopsies were performed at +14 and +30 d. Feeding AAC led to greater PMNL expression of genes related with inflammation response(DDX58), oxidative stress response(MPO), eicosanoid metabolism(PLA2G4A and ALOX5AP), transcription regulation(PPARG), and cellular adhesion(TLN1). The upregulation by AAC in endometrium of genes related with inflammation response( TLR2, TLR4, NFKB1, TNF, IL6, IL1 B, IL10, IL8), prostaglandin synthesis(PTGS2, PTGES), and antioxidant responses(NFE2 L2, SOD1) indicated a faster remodeling of uterine tissue and potentially greater capacity to control a local bacterial invasion.Conclusions: Data indicate that trace mineral supplementation from amino acid complexes improves PMNL activity and allows the prompt recovery of uterine tissue during early lactation. As such, the benefits of complexed trace minerals extend beyond an improvement of liver function and productive performance.

A dynamic model of once-daily 5-aminosalicylic acid predicts clinical efficacy

New once daily mesalamine formulations may improve adherence to medication usage.Response to Asacol and other forms of 5-aminosalicyclic acid(5-ASA)is better correlated with tissue concentrations and best predicted by concentrations of the drug within the lumen of the colon.Our group used computer simulation to predict colonic 5-ASA levels after Asacol administration.In our study,the model simulated Asacol distribution in the healthy colon,and during quiescent and active ulcerative colitis.An Asacol dosage of 800 mg,threetimes a day,was compared to 2400 mg given once a day.Under ideal conditions,the predicted maximum drug in the total colon and individual colonic segments over 100 d differed by less than 3%between single and multiple doses.Despite changes in motility and defection rates,the predicted maximum and average 5-ASA concentrations in the total colon and individual colonic segments differed by less than 10%between dosing regimens.Asymmetric distribution of 5-ASA in the colon was influenced by frequency of bowel movements and colonic transit rate.In active colitis,sigmoid 5-ASA concentration becomes negligible.Our model supports once daily administration of Asacol as standard treatment for ulcerative colitis.

Supplemental Smartamine M in higher-energy diets during the prepartal period improves hepatic biomarkers of health and oxidative status in Holstein cows

Background： Feeding higher-energy prepartum is a common practice in the dairy industry. However, recent data underscore how it could reduce performance, deepen negative energy balance, and augment inflammation and oxidative stress in fresh cows. We tested the effectiveness of rumen-protected methionine in preventing the negative effect of feeding a higher-energy prepartum. Multiparous Holstein cows were fed a control lower-energy diet（CON, 1.24 Mcal/kg DM; high-straw） during the whole dry period（-50 d）, or were switched to a higher-energy（OVE, 1.54 Mcal/kg DM）, or OVE plus Smartamine M（OVE ＋ SM; Adisseo NA） during the last 21 d before calving.Afterwards cows received the same lactation diet（1.75 Mcal/kg DM）. Smartamine M was top-dressed on the OVE diet（0.07% of DM） from -21 through 30 d in milk（DIM）. Liver samples were obtained via percutaneous biopsy at -10, 7 and 21 DIM. Expression of genes associated with energy and lipid metabolism, hepatokines, methionine cycle, antioxidant capacity and inflammation was measured.Results： Postpartal dry matter intake, milk yield, and energy-corrected milk were higher in CON and OVE ＋ SM compared with OVE. Furthermore, milk protein and fat percentages were greater in OVE ＋ SM compared with CON and OVE. Expression of the gluconeogenic gene PCK1 and the lipid-metabolism transcription regulator PPARA was again greater with CON and OVE ＋ SM compared with OVE. Expression of the lipoprotein synthesis enzyme MTTP was lower in OVE ＋ SM than CON or OVE. Similarly, the hepatokine FGF21, which correlates with severity of negative energy balance, was increased postpartum only in OVE compared to the other two groups. These results indicate greater liver metabolism and functions to support a greater production in OVE ＋ SM. At 7 DIM, the enzyme GSR involved in the synthesis of glutathione tended to be upregulated in OVE than CON-fed cows, suggesting a greater antioxidant demand in overfed cows. Feeding OVE ＋ SM resulted in lower similar expression of GSR compared with CON. Expression of the methionine cycle enzymes SAHH and MTR, both of which help synthesize methionine endogenously, was greater prepartum in OVE ＋ SM compared with both CON and OVE, and at 7 DIM for CON and OVE ＋ SM compared with OVE, suggesting greater Met availability. It is noteworthy that DNMT3A, which utilizes S-adenosylmethionine generated in the methionine cycle, was greater in OVE and OVE ＋ SM indicating higher-energy diets might enhance DNA methylation, thus, Met utilization.Conclusions： Data indicate that supplemental Smartamine M was able to compensate for the negative effect of prepartal energy-overfeeding by alleviating the demand for intracellular antioxidants, thus, contributing to the increase in production. Moreover Smartamine M improved hepatic lipid and glucose metabolism, leading to greater liver function and better overall health.

Expression of fatty acid sensing G-protein coupled receptors in peripartal Holstein cows

Background： G-protein coupled receptors（GPCR）, also referred as Free Fatty Acid Receptors（FFAR）, are widely studied within human medicine as drug targets for metabolic disorders. To combat metabolic disorders prevalent in dairy cows during the transition period, which co-occur with negative energy balance and changes to lipid and glucose metabolism, it may be helpful to identify locations and roles of FFAR and other members of the GPCR family in bovine tissues.Results： Quantitative RT-PCR（qPCR） of subcutaneous adipose, liver, and PMNL samples during the transition period（-10, ＋7, and ＋20 or ＋30 d） were used for expression profiling of medium-（MCFA） and long-chain fatty acid（LCFA）receptors GPR120 and GPR40, MCFA receptor GPR84, and niacin receptor HCAR2/3. Adipose samples were obtained from cows with either high（HI; BCS ≥ 3.75） or low（LO; BCS ≤ 3.25） body condition score（BCS） to examine whether FFAR expression is correlated with this indicator of health and body reserves. Supplementation of rumen-protected methionine（MET）, which may improve immune function and production postpartum, was also compared with unsupplemented control（CON） cows for liver and blood polymorphonuclear leukocytes（PMNL） samples. In adipose tissue, GPR84 and GPR120 were differentially expressed over time, while GPR40 was not expressed; in PMNL, GPR40 was differentially expressed over time and between MET vs. CON, GPR84 expression differed only between dietary groups, and GPR120 was not expressed; in liver, GPCR were either not expressed or barely detectable.Conclusions： The data indicate that there is likely not a direct role in liver for the selected GPCR during the transition period, but they do play variable roles in adipose and PMN. In future, these receptors may prove useful targets and/or markers for peripartal metabolism and immunity.

Maternal dietary deficiencies in folic acid or choline worsen stroke outcomes in adult male and female mouse offspring

Maternal one-carbon metabolism plays an important role in early life programming.There is a well-established connection between the fetal environment and the health status of the offspring.Howeve r,there is a knowledge gap on how maternal nutrition impacts stro ke outcomes in offspring.The aim of our study was to investigate the role of maternal dietary deficiencies in folic acid or choline on stroke outcomes in 3-month-old offspring.Adult female mice were fed a folic acid-deficient diet,choline-deficient diet,or control diet 4 weeks before pregnancy.They we re continued on diets during pregnancy and la ctation.Male and female offspring were weaned onto a control diet and at 2 months of age were subjected to ischemic stroke within the sensorimotor cortex via photothrombotic damage.Mothers maintained on either a folic acid-deficient diet or choline-deficient diet had reduced levels of S-adenosylm ethionine in the liver and S-adenosylhomocysteine in the plasma.After ischemic stro ke,motor function was impaired in 3-month-old offspring from mothers receiving either a folic acid-deficient diet or choline-deficient diet compared to the animals receiving a control diet.In brain tissue,there was no difference in ischemic damage volume.When protein levels were assessed in ischemic brain tissue,there were lower levels of active caspase-3 and hypoxia-inducible factor 1α in males compared to females and betaine levels were reduced in offspring from the mothers receiving a choline-deficient diet.Our results demonstrate that a deficient maternal diet at critical time points in neurodevelopment results in worse stro ke outcomes.This study emphasizes the importance of maternal diet and the impact it can have on offspring health.

The LXRB-SREBP1 network regulates lipogenic homeostasis by controlling the synthesis of polyunsaturated fatty acids in goat mammary epithelial cells

Background:In rodents,research has revealed a role of liver X receptors(LXR) in controlling lipid homeostasis and regulating the synthesis of polyunsaturated fatty acids(PUFA).Recent data suggest that LXRB is the predominant LXR subtype in ruminant mammary cells,but its role in lipid metabolism is unknown.It was hypothesized that LXRB plays a role in lipid homeostasis via altering the synthesis of PUFA in the ruminant mammary gland.We used overexpression and knockdown of LXRB in goat primary mammary epithelial cells(GMEC) to evaluate abundance of lipogenic enzymes,fatty acid profiles,content of lipid stores and activity of the stearoyl-Co A desaturase(SCD1) promoter.Results:Overexpression of LXRB markedly upregulated the protein abundance of LXRB while incubation with si RNA targeting LXRB markedly decreased abundance of LXRB protein.Overexpression of LXRB plus T0901317(T09,a ligand for LXR) dramatically upregulated SCD1 and elongation of very long chain fatty acid-like fatty acid elongases 5–7(ELOVL 5–7),which are related to PUFA synthesis.Compared with the control,cells overexpressing LXRB and stimulated with T09 had greater concentrations of C16:0,16:1,18:1n7,18:1n9 and C18:2 as well as desaturation and elongation indices of C16:0.Furthermore,LXRB-overexpressing cells incubated with T09 had greater levels of triacylglycerol and cholesterol.Knockdown of LXRB in cells incubated with T09 led to downregulation of genes encoding elongases and desaturases.Knockdown of LXRB attenuated the increase in triacylglycerol and cholesterol that was induced by T09.In cells treated with dimethylsulfoxide,knockdown of LXRB increased the concentration of C16:0 at the expense of C18:0,while a significant decrease in C18:2 was observed in cells incubated with both si LXRB and T09.The abundance of sterol regulatory element binding transcription factor 1 precursor(p SREBP1) and its mature fragment(n SREBP1) was upregulated by T09,but not LXRB overexpression.In the cells cultured with T09,knockdown of LXRB downregulated the abundance for p SREBP1 and n SREBP1.Luciferase reporter assays revealed that the activities of wild type SCD1 promoter or fragment with SREBP1 response element(SRE) mutation were decreased markedly when LXRB was knocked down.Activity of the SCD1 promoter that was induced by T09 was blocked when the SRE mutation was introduced.Conclusion:The current study provides evidence of a physiological link between the LXRB and SREBP1 in the ruminant mammary cell.An important role was revealed for the LXRB-SREBP1 network in the synthesis of PUFA via the regulation of genes encoding elongases and desaturases.Thus,targeting this network might elicit broad effects on lipid homeostasis in ruminant mammary gland.

Maternal body condition during latepregnancy is associated with in utero development and neonatal growth of Holstein calves

Background:Nutritional management in the dry period can alter body condition score(BCS)in dairy cows,a subjective measure of body fat.As such,differences in BCS during late-pregnancy not only mirror nutrient utilization by fat depots,but also can play important roles on the metabolic and hormonal environment.We investigated the association between cow BCS during late-pregnancy on developmental parameters and blood variables of neonatal calves.Forty-nine multiparous Holstein cows were retrospectively divided by prepartal BCS into normal BCS≤3.25(NormBCS;3.02±0.17,n=30)or high BCS≥3.75(HighBCS;3.83±0.15,n=19)groups.Plasma samples were collected from cows at−10 d relative to parturition.Body weight,hip and wither height,hip width and body length were measured at birth and weekly through weaning(42 d of age)and until 9 weeks of age.Calf blood samples were collected from the jugular vein at birth(before receiving colostrum,0 d),24 h after first colostrum and at 7,21,42 and 50 d of age.The data were subjected to ANOVA using the mixed procedure of SAS.The statistical model included day,BCS,and their interactions.Results:Dry matter intake(kg/d or%of body weight)during the last 4 weeks of pregnancy was lower(P≤0.06)in HighBCS cows.Plasma concentrations of fatty acids,ceruloplasmin,and nitric oxide were greater overall(P<0.05)at d−10 prior to calving in HighBCS cows,and they tended(P=0.08)to have greater concentrations of reactive oxygen metabolites.Birth body weight was lower(P=0.03)in calves born to dams with HighBCS.In addition,plasma concentrations of fatty acids,albumin and urea(P<0.05)were greater in those calves.Although calves born to cows with HighBCS maintained a lower postnatal body weight(P=0.04),hip and wither height,hip width,and body length,there was no difference(P>0.05)in daily starter intake and average daily gain due to maternal BCS.Conclusions:Overall,results highlight an association between BCS during late-gestation on in utero calf development and postnatal growth.A high maternal BCS during late-gestation was associated with lower calf body weights,which could be due to lower maternal intakes and a state of inflammation and metabolic stress.