HC Diet Research Articles

Comparative metabolome analysis of ruminal changes in Holstein dairy cows fed low- or high-concentrate diets

Currently, information on the comprehensive changes in the ruminal metabolites of dairy cows fed high-concentrate diet is limited. This study aimed to compare the composition of whole-ruminal metabolites in dairy cows that were fed a low concentrate diet or a high concentrate diet using modern metabolome analysis. Cows were fed a low-concentrate diet (LC; 40% concentrate feeds, dry matter (DM) basis) or a high-concentrate diet (HC; 70% concentrate feeds, DM basis). GC/MS was used to analyze rumen fluid samples. As compared with the LC group, HC diet significantly increased the concentration of bacterial degradation products (included xanthine, hypoxanthine, uracil, etc.), some toxic compounds (included lipopolysaccharide, biogenic amines, ethanolamine, etc.) and 15 amino acids (included alanine, leucine, glycine, etc.). The enrichment analysis of differentially expressed metabolites indicated that three pathways, including aminoacyl-tRNA biosynthesis; phenylalanine, tyrosine, and tryptophan biosynthesis; and valine, leucine and isoleucine biosynthesis, were significantly enriched after the diet treatments. Correlation network analysis revealed that HC diets altered the ruminal metabolic pattern, and the metabolites in the HC group were more complicated than those in the LC group. The correlations between ruminal metabolites and blood parameters were mainly centralized in the ruminal metabolites and albumin (40 metabolites), followed by globulin (18 metabolites) and total protein (6 metabolites). These findings revealed that HC feeding altered the concentrations of ruminal metabolites as well as the metabolic pattern, and the rumen metabolism could be reflected by blood metabolism to a certain degree.

Influence of malic acid-heat treatment for protecting sunflower protein against ruminal degradation on in vitro methane production: A comparison with the use of malic acid as an additive

Batch cultures of rumen microorganisms were used to compare the fermentation of sunflower seed (SS) and meal (SM) treated with malic acid and heat (MAH; 150°C for 1h) to protect the protein from ruminal degradation with that of the untreated samples supplemented the same amount of malate (as additive), either as free acid (MA) or disodium malate (DSM). As previous studies have shown the influence of donors’ diet on in vitro fermentation, cultures were inoculated with ruminal fluid from four sheep fed a medium-concentrate diet (MC; 50:50 hay:concentrate) or a high-concentrate diet (HC; 15:85 barley straw:concentrate) in a cross-over experimental design. Cultures were sampled at 6 and 16.5h of fermentation. In 16.5h incubations, MAH treatment reduced (P<0.001) CH4 production and ammonia-N concentrations for both substrates (by 60.3 and 45.3% for SS and by 23.7 and 17.2% for SM substrate, respectively). Whereas the MAH treatment reduced total volatile fatty (VFA) production by 16.5% for SS substrate, no effects were observed for SM substrate (P=0.441). For both substrates, adding the same amount of MA to untreated samples (as a feed additive) resulted in greater CH4 production and ammonia-N concentrations compared with the MAH treatment. A positive relationship (P<0.001) was observed between the concentrations of NH3N and CH4 production at both incubation times for both substrates. Compared with the untreated feeds, MA and DSM did not affect (P>0.05) CH4 production for either substrate, but reduced ammonia-N concentrations (13.8%; P=0.004) for SM substrate. Compared with DSM, MA treatment resulted in greater (P<0.05) production of gas, butyrate, isovalerate and valerate and tended (P<0.10) to greater total VFA, and propionate production for SS substrate at 6h incubation, but most differences disappeared after 16.5h incubation. For some fermentation parameters, the response to MAH treatment was more pronounced by using fluid from sheep fed the HC diet than that from MC-fed sheep. In conclusion, the MAH treatment was more effective at reducing CH4 emissions and NH3N concentrations than the supplementation of malic acid or disodium malate as a feed additive.

Impact of diet composition and temperature-humidity index on water and dry matter intake of high-yielding dairy cows.

The temperature-humidity index (THI) is widely used to characterize heat stress in dairy cattle. Diet composition is known to induce variation in metabolic-associated heat production. However, the relationships between THI and diet are poorly characterized with regard to performance and intake behaviour. Therefore, the objectives were to evaluate the impact of THI on water intake (WI), dry matter intake (DMI) and the frequency of drinking and feeding bouts in lactating dairy cows offered four dietary treatments: each contained 20% grass silage and additionally (i) 20% maize silage, 60% concentrate (M-HC); (ii) 60% maize silage, 20% concentrate (M-LC); (iii) 20% pressed beet pulp silage, 60% concentrate (BPS-HC); or (iv) 60% pressed beet pulp silage, 20% concentrate (BPS-LC) (DM basis). Individual WI and DMI were recorded from April to July 2013. Furthermore, dietary effects on milk production and reticular pH were estimated. Milk yield was lowest for M-LC, while energy-corrected milk was similar for all diets. Milk fat percentage was higher and milk protein amount lower for cows offered both LC diets. Reticular pH below 6.3, 6.0 and 5.8 lasted longest for BPS-LC. WI was higher for HC diets. However, the frequency of drinking bouts was not influenced by the ration. Lower DMI occurred for BPS-LC compared to M-LC. Frequency of feeding bouts was significantly higher for LC diets. THI was significantly related to WI, DMI as well as drinking and feeding bouts. Per increasing THI, WI increased slightly more for LC diets and DMI decreased more for HC diets. Frequency of drinking bouts increased slightly higher for BPS rations per rising THI, while the decrease in feeding bouts was highest for M-HC. In conclusion, TMR composition and moderate heat stress impacted WI and DMI of dairy cows, while both dietary energy density and ruminal filling might intensify the THI impact.

Glyceroneogenesis in the hepatopancreas of the crab Neohelice granulata: Diet, starvation and season effects.

We determined the activity of glyceroneogenesis from [2-14C]-pyruvate, the phosphoenolpyruvate carboxykinase activity, [2-14C]-pyruvate oxidation and total lipid levels in the hepatopancreas of the crab Neohelice granulata fed with a carbohydrate-rich (HC) diet or a high-protein (HP) diet and then subjected to 5weeks of starvation, in summer and winter, to determine whether the seasonal adjustments of lipid metabolism to food scarcity are modulated by the composition of the diet previously given to the crabs. The results demonstrated that glyceroneogenesis is an active pathway in N. granulata hepatopancreas, and is regulated by seasonal variations, diet composition and starvation. This study showed that in summer the increase in the hepatopancreas glyceroneogenesis activity is among the strategies used by N. granulata fed an HP diet, to maintain the triglyceride/fatty acid cycle during starvation, a normal condition in the biological cycle of this crab. However, the administration of an HC diet reduced the glyceroneogenesis capacity in response to starvation in summer. In winter, the decrease in the glyceroneogenesis capacity in both fed (HP and HC diets) and starved crabs seems to be a strategy to reduce energy consumption and/or requirement. In contrast to the summer results, the incorporation of [2-14C]-pyruvate into 14CO2 was markedly higher in both diet (HC and HP) groups and in starved crabs during the winter. Four decades after the first study describing the glyceroneogenesis pathway in rat white adipose tissue, this pathway is evidenced for the first time in a crustacean.

&lt;b&gt;Ruminal fermentation of Nellore steers fed crude glycerine replacing starch vs. fibre-based energy ingredient in low or high concentrate diets

Twelve ruminally cannulated steers (401.0 ± 41.5 kg) and 24 mo were used in a replicated arrangement truncated Latin Square with six animals in six treatments and four periods to evaluate the effect of crude glycerine (CG; 80.3% of glycerol) with starch or fiber-based energy ingredients in the concentrate on DMI, DM (DMD) and NDF digestibility (NDFD) and ruminal parameters. Experimental periods were 19 days (14 days for adaptation and 5 days to sampling). Diets were: CO - without CG and corn as ingredient of concentrate; CGC - inclusion of CG (10% of DM) with corn in the concentrate; and CGSH - inclusion of CG (10% of DM) with soybean hulls (SH) in the concentrate. All three diets were offered at low (LC) or high (HC) concentrate level, CL (40 or 60%). Animals fed LC or HC diets had similar DMI, DMD and NDFD. Animals fed diets with CG associated with corn or SH had higher propionate concentrations and lower A:P ratio. Diets with HC increase the propionate but do not affect the NDFD. CG (10% of DM) can be used to replace corn or SH in diets with 40 or 60% of concentrate, without affect NDFD.

Post-weaning and whole-of-life performance of pigs is determined by live weight at weaning and the complexity of the diet fed after weaning.

The production performance and financial outcomes associated with weaner diet complexity for pigs of different weight classes at weaning were examined in this experiment. A total of 720 weaner pigs (360 entire males and 360 females) were selected at weaning (27 ± 3 d) and allocated to pens of 10 based on individual weaning weight (light weaning weight: pigs below 6.5 kg; medium weaning weight: 6.5 to 8 kg; heavy weaning weight: above 8.5 kg). Pens were then allocated in a 3 × 2 × 2 factorial arrangement of treatments with the respective factors being weaning weight (heavy, medium and light; H, M and L, respectively), weaner diet complexity (high complexity/cost, HC; low complexity/cost, LC), and gender (male and female). Common diets were fed to both treatment groups during the final 4 weeks of the weaner period (a period of 39 days). In the first 6 d after weaning, pigs offered the HC diets gained weight faster and used feed more efficiently than those offered the LC diets (P = 0.031). Pigs fed a HC diet after weaning tended to be heavier at the sale live weight of 123 d of age compared with pigs fed the LC diet (P = 0.056). There were no other main effects of the feeding program on growth performance through to slaughter. Weaning weight had a profound influence on lifetime growth performance and weight at 123 d of age, with H pigs at weaning increasing their weight advantage over the M and L pigs (101.3, 97.1, 89.6 kg respectively, P < 0.001). Cost-benefit analyses suggested there was a minimal benefit in terms of cost per unit live weight gain over lifetime when pigs were offered a HC feeding program to L, with a lower feed cost/kg gain. The results from this investigation confirm the impact of weaning weight on lifetime growth performance, and suggest that a HC feeding program should be focused on L weaner pigs (i.e., weaning weight less than 6.5 kg at 27 d of age) in order to maximise financial returns.

Long-term administration of benfotiamine benefits the glucose homeostasis of juvenile blunt snout bream Megalobrama amblycephala fed a high-carbohydrate diet

This study aimed to investigate the effects of benfotiamine on growth performance, glucose homeostasis and intermediary glucose metabolism of blunt snout bream (Megalobrama amblycephala). A total of 240 fish (average body weight: 24.24±0.05g) were randomly distributed into 12 tanks and fed three diets (control, diet with 30% carbohydrate; HC, diet with 43% carbohydrate; HCB, HC diet supplemented with 2.85mg/kg benfotiamine) for 12weeks. The results indicated that fish fed the HCB diet had a lower specific growth rate and feed conversion ratio compared to those fed the control and HC diets, whereas protein efficiency ratio, nitrogen retention efficiency and energy retention efficiency showed an opposite trend. Additionally, fish offered HCB diet showed relatively high values of whole-body lipid contents, tissue glycogen and lipid contents and plasma insulin levels, whereas plasma levels of glucose, advanced glycation end products and glycated serum protein significantly decreased compared with other treatments. Furthermore, benfotiamine improved the glucose tolerance of fish fed HC diet after a glucose load. High carbohydrate intake significantly up-regulated the mRNA levels of glucokinase, pyruvate kinase, glucose transporter 2, glycogen synthase, fatty acid synthetase and glucose-6-phosphate dehydrogenase, whereas the opposite was true for phosphoenolpyruvate carboxykinase, fructose-1,6-biphosphatase, glucose-6-phosphatase, carnitine palmitoyl transferase I and acyl-CoA oxidase. This also held true for fish fed HCB diet except for fatty acid oxidation process which was further enhanced. Overall, these findings suggested that benfotiamine benefited the glucose metabolism of Megalobrama amblycephala fed high-carbohydrate diet through the stimulation of glycolysis, glycogenesis, lipogenesis and fatty acid oxidation coupled with the depression of gluconeogenesis.Statement of relevanceThis study investigated the effects of dietary benfotiamine supplementation on growth performance, glucose homeostasis and intermediary metabolism of blunt snout bream. The data obtained here will facilitate our understanding of the potential mechanisms concerning the beneficial effects of benfotiamine on the carbohydrate metabolism of fish. It is also helpful for the development of low-protein and high-energy feed for fish.

Glucomannan- and glucomannan plus spirulina-enriched pork affect liver fatty acid profile, LDL receptor expression and antioxidant status in Zucker fa/fa rats fed atherogenic diets

ABSTRACTWe evaluated the effects of glucomannan or glucomannan plus spirulina-restructured pork (RP) on liver fatty acid profile, desaturase/elongase enzyme activities and oxidative status of Zucker fa/fa rats for seven weeks. Control (C), glucomannan (G) and glucomannan/spirulina (GS)-RP; HC (cholesterol-enriched control), HG and HGS (cholesterol-enriched glucomannan and glucomannan/spirulina-RP) experimental diets were tested. Increased metabolic syndrome markers were found in C, G and GS rats. Cholesterol feeding increased liver size, fat, and cholesterol and reduced antioxidant enzyme levels and expressions. Cholesterolemia was lower in HG and HGS than in HC. GS vs. G showed higher stearic but lower oleic levels. SFA and PUFA decreased while MUFA increased by cholesterol feeding. The arachidonic/linoleic and docosahexaenoic/alpha-linolenic ratios were lower in HC, HG, and HGS vs. C, G, and GS, respectively, suggesting a delta-6-elongase-desaturase system inhibition. Moreover, cholesterol feeding, mainly in HGS, decreased low-density-lipoprotein receptor expression and the delta-5-desaturase activity and increased the delta-9-desaturase activity. In conclusion, the liver production of highly unsaturated fatty acids was limited to decrease their oxidation in presence of hypercholesterolaemia. Glucomannan or glucomannan/spirulina-RP has added new attributes to their functional properties in meat, partially arresting the negative effects induced by high-fat-high-cholesterol feeding on the liver fatty acid and antioxidant statuses.

Citrus junos Tanaka peel ameliorates hepatic lipid accumulation in HepG2 cells and in mice fed a high-cholesterol diet.

Background Citrus junos Tanaka (yuja), a yellow-coloured citrus fruit has traditionally been consumed in Korea, Japan, and China and has been found effective in preventing certain diseases. However, the inhibitory effect of yuja on hepatic lipid accumulation has not been clearly elucidated thus far.MethodsThe inhibitory effect of yuja on hepatic lipid accumulation was investigated in both cell culture and mouse models. We investigated the inhibitory effect of ethanol extract of yuja peel (YE) using HepG2 cells. We next confirmed the effect of YE in mice fed a high cholesterol diet. Animals were divided into 4 groups (n = 8): a normal diet group (ND), a high-cholesterol diet group (HC), high-cholesterol diet plus 1% YE (YL), high-cholesterol diet plus 5% YE (YH).ResultSeventy percent ethanolic extracts of yuja peel (YE) reduced oleic acid-induced hepatic lipid accumulation in HepG2 cells. Treatment with YE at 100, 200 μg/mL up-regulated expression levels of cholesterol metabolism-related proteins such as AMPK, ACC, PPAR-α, and CPT1 and down-regulated the expression of 3-hydroxy-3-methylglutaryl coenzyme A reductase. The hypocholesterolemic effect of YE was further confirmed in mice fed a high-cholesterol diet. Compared to ND (normal diet) mice, HC (high-cholesterol diet) mice showed increased body weight, liver fat content, liver weight, and content of total cholesterol and low-density lipoprotein (LDL) cholesterol. On the contrary, administrations of YL (HC + 1% YE) or YH (HC + 5% YE) significantly reduced body weight, liver fat content, liver weight, total cholesterol, and LDL cholesterol compared to those of only HC fed mice group. As a result of in vitro data, protein expressions of PPAR-α and CPT1 were induced in mice fed YE diet compared to HC diet but HMGCR expression was decreased.ConclusionsYuja peel ameliorates hepatic lipid accumulation in both cell culture and mouse models and therefore, could serve as a useful supplement for hypercholesterolemia.

Evaluating the effects of refined carbohydrate and fat diets with acute ethanol consumption using a mouse model of alcoholic liver injury

Alcoholism is a multifactorial and complex disorder responsible for 5.9% of deaths worldwide. Excessive consumption of ethanol (Et-OH) induces alcoholic liver disease (ALD), a condition comprising a spectrum of clinical signs and morphological changes, ranging from fatty liver (steatosis) to more severe forms of chronic liver injury. Secondary cofactors, such as nutritional and hepatotoxic comorbid conditions, can also contribute to liver disease development. Here we investigated the effects in the progression of ALD following short-term exposure to diet high in refined carbohydrates (HC), a high-sugar and -butter (HSB) hypercaloric diet and acute Et-OH consumption. HSB diet increased the body weight (BW) and adiposity independently of acute Et-OH consumption. HC diet did not affect BW but increased the adiposity, while acute Et-OH alone did not affect BW and adiposity. All groups of mice developed steatosis except the control group. Exposure to acute Et-OH and HSB diet increased the number of neutrophils and macrophages, and apoptosis in the liver. This combination also increased the number of circulating neutrophils and reduced mononuclear cells in the blood. Thus, short-term exposure to HSB diet and acute Et-OH intake is linked to increased liver injury. These findings offer important clues to understand the hepatic injuries associated with short exposure to hypercaloric diets and acute Et-OH.

Chronic Moderate Alcohol Intakes Accelerate SR-B1 Mediated Reverse Cholesterol Transport.

Cholesterol is essential for all animal life. However, a high level of cholesterol in the body is strongly associated with the progression of various severe diseases. In our study, the potential involvement of alcohol in the regulation of high density lipoprotein (HDL) receptor scavenger receptor class B and type I (SR-B1)-mediated reverse cholesterol transport was investigated. We separated male C57BL/6 mice into four diets: control, alcohol, Control + HC and alcohol + HC. The SR-B1 level and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate- high- density lipoprotein (DiI-HDL) uptake were also measured in AML12 cells and HL7702 cells treated with alcohol. The control + HC diet led to increased hepatic triglyceride and cholesterol levels while alcohol + HC led no significant change. Compared with that of the control group, the SR-B1 mRNA level was elevated by 27.1% (P < 0.05), 123.8% (P < 0.001) and 343.6% (P < 0.001) in the alcohol, control + HC and alcohol + HC groups, respectively. In AML12 and HL7702 cells, SR-B1 level and DiI-HDL uptake were repressed by SR-B1 siRNA or GW9662. However, these effects were reversed through alcohol treatment. These data suggest that a moderate amount of alcohol plays a novel role in reverse cholesterol transport, mainly mediated by PPARγ and SR-B1.

Carbohydrate-enriched diet predispose to anxiety and depression-like behavior after stress in mice

Objectives: Obesity is a chronic disease frequently associated with serious co-morbidities, such as diabetes type II, metabolic syndrome, and psychiatric disorders. Little is known, however, regarding the behavioral consequences of modified diet constituents and the propensity to development of stress related disorders. Thus, the aim of this study was to verify whether chronic exposure to a normocaloric/high-carbohydrate diet will modify the animal's behavior after different stressful stimuli.Methods: BALB/c mice were fed for 12 weeks with a standard chow diet or high refined carbohydrate-containing diet (HC). Following this period, independent groups of animals were exposed to different stress paradigms: 1 – two hours of restraint stress followed by exposure to the Elevated Plus Maze test (EPM) 24 hours later; 2 – The contextual fear conditioning (CFC) test and 3 – the tail suspension test (TST).Results: Despite no change on total body weight, animals fed with HC diet showed increase in serum leptin levels and higher adiposity compared to diet control group. In behavioral tests, animals from HC diet group displayed reduction in the percentage of entries into the open arms of the EPM, evaluated 24 hours after restraint stress, suggesting an anxiogenic-like effect. It is also observed increase in aversive memory in the CFC test and depressive-like behavior in TST.Discussion: Our results suggest that a moderate obesity, induced by high refined carbohydrate diet, may facilitate the development of anxiety and depressive-like behaviors after the stress. The mechanisms responsible for such effects remain to be elucidated.

Gestational Exposure to Maternal Obesity Influences β3‐Adrenergic Agonist Induced Beiging of White Adipose Tissue in Offspring

Maternal obesity is now recognized as an important contributing factor to the currently unfolding epidemic of obesity. In animal models, maternal obesity and exposure to high‐fat diets in utero, unequivocally predisposes the offspring to greater weight gain, metabolic dysfunction and a host of other related comorbidities, such as fatty liver disease. Our previous work in both rat and mouse models of obesity showed that offspring of obese dams are hyper‐responsive to an obesogenic challenge post‐weaning, and gain greater weight and fat mass. In part this is associated with decreased energy expenditure, impaired expression of mitochondrial OXPHOS and lipid oxidative genes and lower expression uncoupling proteins in liver, skeletal muscle and white adipose tissue. Nonetheless, the consequences of maternal obesity on programming of adipose tissue development, especially brown/beige adipose tissue development are unknown. In this report, we investigated role of maternal obesity on beige fat development in offspring following β3‐adrenergic agonist. Female C57BL6/J mice were fed diets with either low (10%) or HFD (45% of calories) for 12 wk starting at 5 wk of age. Dams were mated with males on control diets and provided access to respective diets during pregnancy and lactation. At weaning, offspring were weaned onto control and HFD, creating four groups (CC, CH, HC, HH, denoting maternal and offspring diets respectively). Male offspring from the four groups were followed till 20 wk of age. HFD‐fed offspring from HF dams (HH) gained significantly (p&lt;0.05) greater body weights than offspring from lean dams (CH). At 20 wk of age, offspring were treated with either vehicle or β3‐adrenergic receptor agonist CL‐316,243 (1 mg/kg, ip) for one week (N=4–5 each). CL‐316,243 is a well established activator of beige adipocyte formation. Twenty weeks of diet intervention lead to significantly consistent increased weight gain in HH compared to other groups. One week of CL injection towards to end of study did not affect the weight gain. There were no significant differences in overall body weight between vehicle and CL treated animals. Gene expression analysis of inguinal adipose tissue depot (iWAT) revealed significant differences in vehicle and CL treated groups. Expression of brown‐fat specific genes such as Ucp1, Cidea and Pgc1a was affected by maternal HF diet feeding in both vehicle and CL treated group. Moreover, maternal obesity hampered the response to CL‐316,243 consistent with programming defect in adipose tissue. Further analysis of developmental aspects of beiging of adipose tissue is under investigation. Our findings indicate that maternal obesity influences beige cell formation in response to classical beiging inducers which might influence modulation of energy expenditure, metabolism and overall risk of obesity.Support or Funding InformationSupport USDA CRIS 6206‐51000‐010‐05S

Maternal Obesity Programs Offspring's Predisposition to Non‐Alcoholic Fatty Liver Disease and Steatohepatitis

Maternal obesity and unbalanced diets during pregnancy are key factors in determining the risk of obesity in later life. Non‐alcoholic fatty liver disease (NAFLD) is an important co‐morbidity associated with obesity and metabolic dysfunction and a precursor to steatohepatitis (NASH). However, the contributions of gestational and early neonatal influences on development of NAFLD and subsequent NASH remain poorly appreciated. Using a model of controlled overfeeding, we have previously demonstrated that in utero exposure to maternal obesity during pregnancy alone leads to metabolic programming of liver, characterized by metabolic inflexibility. Here we examine the hypothesis that maternal over‐nutrition via in utero exposure to high fat diet induces inflammation and leads to exacerbated hepatic responses to post‐natal high fat diets (HFD) and methionine choline deficient (MCD) diets in the offspring. Female C57BL6/J mice were fed diets with either low (10%) or HFD (45% of calories) for 12 wk starting at 5 wk of age. Dams were mated with males on control diets and provided access to respective diets during pregnancy and lactation. At weaning, offspring were weaned onto control and HFD, creating four groups (CC, CH, HC, HH, denoting maternal and offspring diets, respectively). Offspring of obese mice dams (HH n=7) gained greater body weight and fat mass when fed HFD, compared to offspring from lean dam (CH n=11). Serum cholesterol concentrations were higher in HH compared to CH groups. Global hepatic gene expression and microbial ecology of the cecal contents in offspring was studied using RNAseq and 16S rRNA sequencing. Hepatic expression of Mcp1 and MMP13 gene expression was higher in HH compared to CC and CH indicative of inflammation in response to maternal high fat diet. Analysis of microbial OTUs with Multivariate association with linear models (MaAsLin) and LeFSE showed higher abundance of Firmicutes and lower abundance of Bacteriodetes in HH as well as CH compared to CC. To assess the susceptibility of offspring to NASH, 8 wk old offspring in a separate study were challenged with either control or MCD diets for 4 wk. MCD diet led to significant weight reduction irrespective of maternal nutritional background. Serum ALT was modestly higher in offspring from obese dams (HFD/MCD) relative to lean dam cohorts. Offspring of HF dams showed similar inflammatory changes but greater pro‐fibrogenic alterations following MCD diet. MaAsLin and LeFSE analysis revealed association between serum ALT levels, Coprococcus and MCD treatment. Genome‐scale DNA methylation of livers showed alterations in CpGs associated with cell adhesion, cell communication and development with MCD challenge, including genes in the TGF‐β signaling pathways. Overall, our findings indicate that maternal obesity during gestation and lactation detrimentally alters epigenetic and gut microbiome pathways to favor development of fatty liver disease and its progressive sequelae.Support or Funding InformationSupport USDA CRIS 6206‐51000‐010‐05S

The Effects of Dietary Macronutrient Composition on Lipid Metabolism‐Associated Factor Gene Expression in the Adipose Tissue of Chickens are Influenced by 3 Hours of Fasting

Our objective was to determine how dietary macronutrient composition and fasting affect chick adipose tissue physiology during the early post‐hatch period. Broiler chicks were fed one of three isocaloric (3,000 kcal metabolizable energy (ME)/kg) and isonitrogenous (22% crude protein; CP) corn and soybean meal‐based diets after hatch: high carbohydrate (HC; control), high fat (HF; 30% of ME from soybean oil) or high protein (HP; 27% CP). At 4 days post hatch, chicks were fasted for 3 hours, and clavicular, gizzard and subcutaneous adipose depots were collected from fasted and fed chicks (n=10 per group). Total RNA was isolated, reverse transcribed and real‐time PCR performed to quantify 1‐acylglycerol‐3‐phosphate O‐acyltransferase 9 (AGPAT9), fatty acid binding protein 4 (FABP4), CCAAT/enhancer‐binding protein alpha (CEBPα), CEBPβ, peroxisome proliferator‐activated receptor gamma (PPARγ), monoglyceride lipase (MGLL), and sterol regulatory element‐binding transcription factor 1 (SREBP‐1) mRNA. The statistical model (within fat depot) included the main effects of diet and fasting, and the interaction. Means were separated within diet. There was greater CEBPα, PPARγ, and SREBP‐1, and less CEBPβ mRNA in clavicular, gizzard and subcutaneous fat in fasted than fed chicks (P &lt; 0.05). In clavicular (P = 0.002) and subcutaneous (P = 0.01) fat, FABP4 expression was lower in chicks fed HP than HF or HC. In clavicular (P = 0.003) and gizzard (P = 0.009) fat, CEPBβ mRNA was greater in HF and HC than HP‐fed chicks. MGLL mRNA was greater (P = 0.007) in clavicular fat of HF than HP‐fed chicks. There were interactions of diet and fasting on PPARγ (P = 0.04) and SREBP‐1 (P = 0.008) in clavicular, PPARγ (P = 0.04) in gizzard, and CEBPα (P = 0.007) in subcutaneous fat. Post‐hoc testing revealed that CEBPα in subcutaneous (P &lt; 0.003) and PPARg mRNA in gizzard (P &lt; 0.03) fat showed a more pronounced decrease in expression during fasting in HC and HF than HP‐fed chicks. In clavicular fat, expression of AGPAT9 (P = 0.001), PPARg (P &lt; 0.0001), and SREBP‐1 (P &lt; 0.0001) mRNA decreased after 3 hours of fasting only in chicks that consumed the HC diet; expression was not affected by food withdrawal in chicks that consumed HF or HP. These data demonstrate that consumption of diets differing in fat and protein content affect gene expression of some lipid metabolism‐associated factors during the first week post‐hatch; in general mRNA was greater in chicks fed HC and HF than the HP diet, with similar effects observed in different anatomical fat depots. Fasting also influenced expression of these factors, in an adipose depot‐dependent manner. These results may have implications for understanding how early‐life nutrition affects development of different adipose tissue depots in birds.Support or Funding InformationFunding for this work was provided in part, by the Virginia Agricultural Experiment Station and the Hatch Program of the National Institute of Food and Agriculture, U.S. Department of Agriculture, and by a John Lee Pratt Fellowship awarded to Guoqing Wang.

An Extract from Prickly Pear Peel (Opuntia ficus-indica) Affects Cholesterol Excretion and Hepatic Cholesterol Levels in Hamsters Fed Hyperlipidemic Diets

The effect of a prickly pear peel extract on cholesterol excretion and plasma and hepatic cholesterol levels in hamsters with induced hypercholesterolemia was investigated. Female and male hamsters were fed for 5 weeks with experimental diets: NC, normal cholesterol diet; HC, hypocholesterolemic diet; HCOp, HC with prickly pear peel extract and HCPh, HC with phytosterols. Male and females hamsters fed HCOp diet increased their neutral fecal sterols (cholesterol + coprastanol) excretion by 77% (p<0.042) and 120% (p<0.003), respectively, compared to counterparts fed HC diet. Plasma total cholesterol was significantly reduced only in male hamsters fed HCPh. The phytosterols content in HCOp diet was almost 5 fold less compared to HCPh, and both treatments have reduced 35% (p<0.0001) hepatic cholesterol compared to hamsters fed the HC diet. Thus, the prickly pear peel extract rich in phytosterols showed a hypocholesterolemic effect. This evidence suggests that the prickly pear byproduct tested herein has potential to be employed as ingredient or supplement to lower cholesterol and prevent cardiovascular diseases. Keywords: Byproduct, hamster, hypocholesterolemic effect, phytosterol, prickly pear peel.

Central Injection of Neuropeptide Y Affects Dietary Macronutrient Composition‐Mediated Changes in Broiler Adipose Tissue mRNA Abundance

Neuropeptide Y (NPY) is a potent orexigenic (hunger increasing) factor that also promotes adipose tissue development in birds and mammals. The purpose of this study was to investigate how dietary macronutrient composition and exogenous NPY affect expression of lipid metabolism‐associated factors in different anatomical adipose (fat) depots of chickens. Chicks were fed one of three isocaloric diets (3,000 kcal metabolizable energy (ME)/kg) and isonitrogenous (22% crude protein; CP) corn and soybean meal‐based diets after hatch: high carbohydrate (HC; control), high fat (HF; 30% of ME from soybean oil) or high protein (HP; 27% CP) post hatch. On day 4 post hatch, 0.2 nmol of NPY or vehicle was injected intra cerebroventricularly (into left lateral ventricle) and abdominal and subcutaneous fat were collected at 1‐hour post injection. Total RNA was isolated, reverse transcribed and real‐time PCR performed to quantify expression of genes associated with adipogenesis: Krüppel‐like factor 7 (KLF7), GATA‐binding protein 2 (GATA 2), CCAAT/enhancer‐binding protein alpha (CEBPα), CEBPβ, peroxisome proliferator‐activated receptor gamma (PPARγ), sterol regulatory element‐binding transcription factor 1 (SREBP‐1), and fatty acid binding protein 4 (FABP4). The statistical model (within fat depot) included the main effects of diet and NPY treatment, and the two‐way interaction. There was an effect of diet (P &lt; 0.05) for all of the genes measured in abdominal and subcutaneous fat, except for CEBPβ in subcutaneous fat. Chicks fed the HC diet expressed more (P &lt; 0.0001) KLF7, CEBPα, CEBPβ, PPARγ, SREBP‐1, and FABP4 in abdominal fat than chicks fed the HF or HP diet. There were interactions of diet and NPY treatment on PPARγ (P = 0.04) and FABP4 (P = 0.03) mRNA in abdominal fat, and CEBPα (P = 0.01), PPARγ (P = 0.001), SREBP‐1 (P = 0.001) and FABP4 (P = 0.001) mRNA in subcutaneous fat. Pairwise comparison tests revealed that CEBPα (P &lt; 0.003), PPARγ (P = 0.007), SREBP‐1 (P = 0.008) and FABP4 (P = 0.001) mRNA showed a more pronounced increase in subcutaneous fat of the HF than HC and HP fed chicks after NPY injection. PPARγ (P = 0.01), SREBP‐1 (P = 0.01) and FABP4 (P = 0.04) also showed a more accentuated decrease in subcutaneous fat of HC than HF and HP fed chicks after NPY injection. PPARγ (P = 0.003) and FABP4 (P = 0.003) in abdominal fat increased more in expression in HF than HC and HP fed chicks after NPY injection. These data demonstrate that consumption of diets differing in fat content affect adipose tissue physiology, with differences in gene expression of some adipogenesis‐associated factors during the first week post‐hatch; in general mRNA was greater in chicks fed HC than the HF or HP diet, with similar effects observed in different anatomical fat depots. Injection of NPY affected expression of these factors in an adipose depot‐dependent manner. These results may have implications for understanding how early‐life nutrition and appetite‐regulatory neuropeptides affect development of different adipose tissue depots in birds.Support or Funding InformationFunding for this work was provided in part, by the Virginia Agricultural Experiment Station and the Hatch Program of the National Institute of Food and Agriculture, U.S. Department of Agriculture, and by a John Lee Pratt Undergraduate Research Scholarship awarded to CW.

Long-term effects of very low-carbohydrate and high-carbohydrate weight-loss diets on psychological health in obese adults with type 2 diabetes: randomized controlled trial.

Very low-carbohydrate, high-fat (LC) diets are used for type 2 diabetes (T2DM) management, but their effects on psychological health remain largely unknown. This study examined the long-term effects of an LC diet on psychological health. One hundred and fifteen obese adults [age: 58.5 ± 7.1 years; body mass index: 34.6 ± 4.3 kg m(-2) ; HbA1c : 7.3 ± 1.1%] with T2DM were randomized to consume either an energy-restricted (~6 to 7 MJ), planned isocaloric LC or high-carbohydrate, low-fat (HC) diet, combined with a supervised exercise programme (3 days week(-1) ) for 1 year. Body weight, psychological mood state and well-being [Profile of Mood States (POMS), Beck Depression Inventory (BDI) and Spielberger State Anxiety Inventory (SAI)] and diabetes-specific emotional distress [Problem Areas in Diabetes (PAID) Questionnaire] and quality of life [QoL Diabetes-39 (D-39)] were assessed. Overall weight loss was 9.5 ± 0.5 kg (mean ± SE), with no difference between groups (P = 0.91 time × diet). Significant improvements occurred in BDI, POMS (total mood disturbance and the six subscales of anger-hostility, confusion-bewilderment, depression-dejection, fatigue-inertia, vigour-activity and tension-anxiety), PAID (total score) and the D-39 dimensions of diabetes control, anxiety and worry, sexual functioning and energy and mobility, P < 0.05 time. SAI and the D-39 dimension of social burden remained unchanged (P ≥ 0.08 time). Diet composition had no effect on the responses for the outcomes assessed (P ≥ 0.22 time × diet). In obese adults with T2DM, both diets achieved substantial weight loss and comparable improvements in QoL, mood state and affect. These results suggest that either an LC or HC diet within a lifestyle modification programme that includes exercise training improves psychological well-being.

Lipopolysaccharide derived from the digestive tract activates inflammatory gene expression and inhibits casein synthesis in the mammary glands of lactating dairy cows.

To meet the nutrition requirements of lactation, dairy cows are usually fed a high concentrate diet (HC). However, high-grain feeding causes subacute ruminal acidosis (SARA), a metabolic disorder that causes milk protein depression. This study aimed to investigate the effect of lipopolysaccharide (LPS) released in the rumen on inflammatory gene expression and casein synthesis in mammary glands of lactating dairy cows fed a HC diet. We found that milk protein was significantly decreased in the HC group after 15 weeks of feeding. Overall, LPS concentrations in the rumen fluid, lacteal artery and vein were increased in the HC group. Transcriptome microarray was used to evaluate alterations in the signaling pathway in mammary glands. Signaling pathways involved in inflammatory responses were activated, whereas those involved in protein synthesis were inhibited in the HC group. mRNA expression involved in inflammatory responses, including that of TLR4, NF-кB and pro-inflammatory genes, was increased in the HC group, while αs1-casein (CSN1S1), β-casein (CSN2), mTOR and S6K gene expression were decreased. Moreover, protein expression was consistent with the corresponding gene expression. After feeding with an HC diet, LPS derived from the rumen increased inflammatory gene expression and inhibited casein synthesis in the mammary glands of lactating dairy cows fed a HC diet.

Comparative proteomic analysis of the effects of high-concentrate diet on the hepatic metabolism and inflammatory response in lactating dairy goats.

BackgroundTo understand the impact of feeding a high-concentrate diet to mid-lactating goats for a long time on liver metabolism and inflammatory response, two dimensional polyacrylamide gel electrophoresis (2-DE) and real-time PCR method were employed to detect proteins differentially expressed in liver and their mRNAs expression in goats fed high concentrate diet (HC) or low concentrate diet (LC). Twelve lactating dairy goats were randomly assigned to either a HC diet group (65 % concentrate of dry matter; n = 6) or a LC diet group (35 % concentrate of dry matter; n = 6) for 10 wk.ResultsTwenty differentially expressed spots (≥2.0-fold changes) in the hepatic tissues were excised and successfully identified using MALDI TOF/TOF. Of these, 8 proteins were up-regulated, while the rest 12 proteins were down-regulated in HC goats compared to LC. Differential expressed proteins including alpha enolase 1 (ENO1), glutamate dehydrogenase 1 (GLUD1), glutathione S-transferase A1 (GSTA1), ATP synthase subunit 5β (ATP5β), superoxide dismutase [Cu-Zn] (SOD1), cytochrom c oxidase subunit Via (COX6A1) and heat shock protein 60 (HSP60) were further verified by real-time PCR and/or western blot at mRNA or protein expression level. Consistent with the 2-DE results, a significant decrease of β-actin protein expression and SOD enzyme activity was observed in liver of HC goats (P < 0.05), while ENO1 protein expression was significantly up-regulated in HC compared to LC goats (P < 0.05) . However, western blot analysis did not show a significant difference of hepatic HSP60 protein between HC and LC group, which did not match the decrease of HSP60 content detected by 2-DE analysis. Real-time PCR showed that glutathione S-transferase P1 (GSTP1) and SOD1 mRNA expression was significantly decreased in liver of HC goats, while cytochrom c oxidase (COX3) and ATPase 8 (ATP8) mRNAs expression were markedly increased compared to LC (P < 0.05). Gene Ontology (GO) analysis revealed that HC diet resulted in altered expression of proteins related to catalytic and mitochondrial metabolism in the liver, and may increase the stress response with up-regulating the expression of differentiation 14 (CD14) cluster and serum amyloid A (SAA) as well as C-reactive protein (CRP) in the liver.ConclusionsThese results suggest that feeding high concentrate diet to lactating goats for 10 wk leads to the activation of the inflammatory response, and decreases the anti-oxidant capacity, and subsequently impairs the mitochondrial function in the liver.Electronic supplementary materialThe online version of this article (doi:10.1186/s40104-016-0065-0) contains supplementary material, which is available to authorized users.