Dose- and substrate-dependent reduction of enteric methane and ammonia by natural additives in vitro.

Ruminants contribute to global warming by emitting greenhouse gasses, particularly methane (CH4) which is a product of rumen fermentation. The use of feed additives able to modulate rumen fermentation is a promising strategy to reduce enteric CH4 and ammonia (NH3) emissions. Among the various strategies investigated, plant secondary metabolites (PSMs) have attracted attention due to their apparent potential to reduce enteric CH4 and NH3 emissions, and it would be possible to use such compounds as feed additives in organic production systems. In an in vitro system simulating rumen fermentation, we have tested the impact of different classes of naturally occurring PSMs; catechin and quercetin (flavonoids), salicylic acid (phenolic acid) and tannic acid (hydrolysable tannin). The PSMs were added to two different basal feeds (maize and grass silages) at three inclusion doses 1.5, 3 and 6% of the feed dry matter (DM). CH4 production was significantly lowered upon addition of quercetin to two basal feeds at doses of 3 and 6%, and this without changes in concentrations of total volatile fatty acid (VFA) produced during fermentation. Quercetin, as the only tested additive, reduced CH4 production, and when added to maize silage and grass silage, the reduction increased linearly with increasing dose, ie., by 51 and 43%, respectively, at a dose of 3% of feed DM and by 86 and 58%, respectively, at a dose of 6% of feed DM. Moreover, quercetin significantly reduced NH3 concentration by >12% at doses of 3 and 6% in feed DM irrespective of the basal feed used as compared to when the basal feeds were incubated alone. Although none of the other additives affected CH4 formation, several additives had significant impacts on concentrations of NH3 and VFAs in the incubated fluid after fermentation. This study demonstrated a dose-dependent ability of quercetin to reduce CH4 emission from rumen fermentation, however, the magnitude of the suppression of CH4 depended on the basal feed. Furthermore, quercetin reduced NH3 concentration irrespective of the basal feed type. These findings encourage to in vivo studies to verify whether quercetin can reduce CH4 emission also in cows.

Methane reduction by quercetin, tannic and salicylic acids: influence of molecular structures on methane formation and fermentation in vitro.

Plant secondary metabolites (PSMs) can potentially reduce ruminal methane formation. However, related to differences in their molecular structures, it is not yet clear what causes an anti-methanogenic effect. In an in vitro system simulating rumen fermentation, we investigated the impact of eight compounds with distinct chemical characteristics (gallic and salicylic acids, tannic acid, catechin, epicatechin, quercetin, rutin, and salicin) when added to a basal feed (maize silage) at a concentration of 12% of the feed dry matter. After 48 h of incubation in buffered rumen fluid, methane production was significantly lowered by quercetin (43%), tannic acid (39%) and salicylic acid (34%) compared to the control (maize silage alone) and without changes in total volatile fatty acid production during fermentation. No other PSM reduced methane formation as compared to control but induced significant differences on total volatile fatty acid production. The observed differences were related to lipophilicity, the presence of double bond and carbonyl group, sugar moieties, and polymerization of the compounds. Our results indicate the importance of distinct molecular structures of PSMs and chemical characteristics for methane lowering properties and volatile fatty acid formation. Further systematic screening studies to establish the structure-function relationship between PSMs and methane reduction are warranted.

Enteric methane emission of dairy cows supplemented with iodoform in a dose-response study.

Enteric methane (CH4) emission is one of the major greenhouse gasses originating from cattle. Iodoform has in studies been found to be a potent mitigator of rumen CH4 formation in vitro. This study aimed to quantify potential of iodoform as an anti-methanogenic feed additive for dairy cows and investigate effects on feed intake, milk production, feed digestibility, rumen microbiome, and animal health indicators. The experiment was conducted as a 4 × 4 Latin square design using four lactating rumen, duodenal, and ileal cannulated Danish Holstein dairy cows. The treatments consisted of four different doses of iodoform (1) 0 mg/day, (2) 320 mg/day, (3) 640 mg/day, and (4) 800 mg/day. Iodoform was supplemented intra-ruminally twice daily. Each period consisted of 7-days of adaptation, 3-days of digesta and blood sampling, and 4-days of gas exchange measurements using respiration chambers. Milk yield and dry matter intake (DMI) were recorded daily. Rumen samples were collected for microbial analyses and investigated for fermentation parameters. Blood was sampled and analyzed for metabolic and health status indicators. Dry matter intake and milk production decreased linearly by maximum of 48% and 33%, respectively, with increasing dose. Methane yield (g CH4/kg DMI) decreased by maximum of 66%, while up to 125-fold increases were observed in hydrogen yield (g H2/kg DMI) with increasing dose of iodoform. Total tract digestibility of DM, OM, CP, C, NDF, and starch were unaffected by treatments, but large shifts, except for NDF, were observed for ruminal to small intestinal digestion of the nutrients. Some indicators of disturbed rumen microbial activity and fermentation dynamics were observed with increasing dose, but total number of ruminal bacteria was unaffected by treatment. Serum and plasma biomarkers did not indicate negative effects of iodoform on cow health. In conclusion, iodoform was a potent mitigator of CH4 emission. However, DMI and milk production were negatively affected and associated with indications of depressed ruminal fermentation. Future studies might reveal if depression of milk yield and feed intake can be avoided if iodoform is continuously administered by mixing it into a total mixed ration.

Gene coexpression network analysis reveals perirenal adipose tissue as an important target of prenatal malnutrition in sheep.

We have previously demonstrated that pre- and early postnatal malnutrition in sheep induced depot- and sex-specific changes in adipose morphological features, metabolic outcomes, and transcriptome in adulthood, with perirenal (PER) as the major target followed by subcutaneous (SUB) adipose tissue. We aimed to identify coexpressed and hub genes in SUB and PER to identify the underlying molecular mechanisms contributing to the early nutritional programming of adipose-related phenotypic outcomes. Transcriptomes of SUB and PER of male and female adult sheep with different pre- and early postnatal nutrition histories were used to construct networks of coexpressed genes likely to be functionally associated with pre- and early postnatal nutrition histories and phenotypic traits using weighted gene coexpression network analysis. The modules from PER showed enrichment of cell cycle regulation, gene expression, transmembrane transport, and metabolic processes associated with both sexes' prenatal nutrition. In SUB (only males), a module of enriched adenosine diphosphate metabolism and development correlated with prenatal nutrition. Sex-specific module enrichments were found in PER, such as chromatin modification in the male network but histone modification and mitochondria- and oxidative phosphorylation-related functions in the female network. These sex-specific modules correlated with prenatal nutrition and adipocyte size distribution patterns. Our results point to PER as a primary target of prenatal malnutrition compared to SUB, which played only a minor role. The prenatal programming of gene expression and cell cycle, potentially through epigenetic modifications, might be underlying mechanisms responsible for observed changes in PER expandability and adipocyte-size distribution patterns in adulthood in both sexes.

Dietary inclusion of methanotrophic microbial cell-derived protein in the early postweaning period sustains growth performance and intestinal health of weaner piglets.

The global demand for sustainably produced protein feeds for animal production is increasing. Methanotrophic bacteria grow on methane and convert it into microbial cell protein (MCP) that has been shown to have high nutritive value for growing pigs. The present aimed to investigate how increasing amounts of MCP in diets fed during the first 15 days after weaning affect the growth performance of piglets from weaning until day 43 postweaning. Furthermore, the effect of MCP on intestinal morphology and histopathology was assessed on day 15 after weaning. During seven consecutive weeks, approximately 480 piglets were recruited for the experiment per batch. The piglets were divided into four groups and housed in eight double pens with 60 piglets per pen. The piglets were fed one of four experimental diets with 0, 3, 6, or 10% of MCP included at the expense of fishmeal and subsequently potato protein for the first 15 days postweaning. Thereafter, all pigs were fed commercial weaner diets in two phases (days 16-30 and days 31-43) until day 43 postweaning. All diets were without medicinal zinc. Feed intake and growth were registered on double pen level during all three phases. On day 15 after weaning, 10 piglets per treatment were randomly selected, autopsied, and sampled for intestinal morphology and histopathology. Daily gain during the first 15 days postweaning tended (P = 0.09) to be affected by the inclusion of MCP in the weaning diet being lowest in the group fed 10% MCP. Treatment did not affect daily feed intake; however, Feed Conversion Ratio (FCR) was significantly affected (P = 0.003) showing the highest FCR in piglets fed 10% MCP. Growth performance was not affected by the experimental treatment during the following phases. In the small intestine, villous height tended (P = 0.09) to show a quadratic response to level of MCP in the diet with the longest villi observed after feeding 6% MCP. Dietary treatment did not affect crypt depth. The villous height to crypt depth (VC) ratio showed a quadratic response to increased dietary inclusion of MCP (P = 0.02) with piglets fed 6% MCP having the highest VC ratio. In conclusion, this study demonstrated that MCP could constitute 6% of diets as-fed (22% of total CP), at the expense of fishmeal and potato protein, for newly weaned piglets without negative effects on growth rates and FCR. The inclusion of MCP in diets for newly weaned piglets could be part of improving the sustainability of pig production.

Impact of dietary fat levels and fatty acid composition on milk fat synthesis in sows at peak lactation.

BACKGROUND: Dietary fat is important for energy provision and immune function of lactating sows and their progeny. However, knowledge on the impact of fat on mammary transcription of lipogenic genes, de novo fat synthesis, and milk fatty acid (FA) output is sparse in sows. This study aimed to evaluate impacts of dietary fat levels and FA composition on these traits in sows. Forty second-parity sows (Danish Landrace × Yorkshire) were assigned to 1 of 5 dietary treatments from d 108 of gestation until weaning (d 28 of lactation): low-fat control diet (3% added animal fat); or 1 of 4 high-fat diets with 8% added fat: coconut oil (CO), fish oil (FO), sunflower oil (SO), or 4% octanoic acid plus 4% FO (OFO). Three approaches were taken to estimate de novo milk fat synthesis from glucose and body fat. RESULTS: Daily intake of FA was lowest in low-fat sows within fat levels (P < 0.01) and in OFO and FO sows within high-fat diets (P < 0.01). Daily milk outputs of fat, FA, energy, and FA-derived carbon reflected to a large extent the intake of those. On average, estimates for de novo fat synthesis were 82 or 194 g/d from glucose according to method 1 or 2 and 255 g de novo + mobilized FA/d according to method 3. The low-fat diet increased mammary FAS expression (P < 0.05) and de novo fat synthesis (method 1; P = 0.13) within fat levels. The OFO diet increased de novo fat synthesis (method 1; P < 0.05) and numerically upregulated mammary FAS expression compared to the other high-fat diets. Across diets, a daily intake of 440 g digestible FA minimized milk fat originating from glucose and mobilized body fat. CONCLUSIONS: Sows fed diets with low-fat or octanoic acid, through upregulating FAS expression, increased mammary de novo fat synthesis whereas the milk FA output remained low in sows fed the low-fat diet or high-fat OFO or FO diets, indicating that dietary FA intake, dietary fat level, and body fat mobilization in concert determine de novo fat synthesis, amount and profiles of FA in milk.

Microbiome Research as an Effective Driver of Success Stories in Agrifood Systems - A Selection of Case Studies.

Increasing knowledge of the microbiome has led to significant advancements in the agrifood system. Case studies based on microbiome applications have been reported worldwide and, in this review, we have selected 14 success stories that showcase the importance of microbiome research in advancing the agrifood system. The selected case studies describe products, methodologies, applications, tools, and processes that created an economic and societal impact. Additionally, they cover a broad range of fields within the agrifood chain: the management of diseases and putative pathogens; the use of microorganism as soil fertilizers and plant strengtheners; the investigation of the microbial dynamics occurring during food fermentation; the presence of microorganisms and/or genes associated with hazards for animal and human health (e.g., mycotoxins, spoilage agents, or pathogens) in feeds, foods, and their processing environments; applications to improve HACCP systems; and the identification of novel probiotics and prebiotics to improve the animal gut microbiome or to prevent chronic non-communicable diseases in humans (e.g., obesity complications). The microbiomes of soil, plants, and animals are pivotal for ensuring human and environmental health and this review highlights the impact that microbiome applications have with this regard.

Untargeted Metabolomics Combined with Solid Phase Fractionation for Systematic Characterization of Bioactive Compounds in Hemp with Methane Mitigation Potential.

This study systematically evaluates the presence of methane mitigating metabolites in two hemp (Cannabis sativa L.) varieties, Futura 75 and Finola. Hemp metabolites were extracted with methanol and fractionated using Solid Phase Extraction (SPE). Extracts, fractions, and the remaining pulp were screened for their methane mitigating potential using an in vitro model of rumen fermentation. The bioactive metabolites were identified with Liquid Chromatography-Mass Spectrometry (LC-MS). When incubated with a standard feed (maize silage), the extract of Futura 75 significantly reduced methane production compared to that of control (without added extract) and without negative effects on feed degradability and volatile fatty acid patterns. The compounds responsible for the methane mitigating effect were assigned to flavonoid glycosides. However, none of the fractions of Futura 75 or the pulp exhibited similar effect on methane emission. Butyric acid concentration in the fermentation inoculum was significantly increased, which could indicate why methane production was higher, when incubated with the fractions and the pulp. The extract of Finola did not show a similar significant effect, however, there was a numerical tendency towards lower methane production. The difference in methane mitigating properties between Cannabis sativa L. Futura 75 and Finola, could be related to the content of bioactive flavonoids.

Supplementation of a lacto-fermented rapeseed-seaweed blend promotes gut microbial- and gut immune-modulation in weaner piglets.

BACKGROUND: The direct use of medical zinc oxide in feed will be abandoned after 2022 in Europe, leaving an urgent need for substitutes to prevent post-weaning disorders. RESULTS: This study investigated the effect of using rapeseed-seaweed blend (rapeseed meal added two brown macroalgae species Ascophylum nodosum and Saccharina latissima) fermented by lactobacilli (FRS) as feed ingredients in piglet weaning. From d 28 of life to d 85, the piglets were fed one of three different feeding regimens (n = 230 each) with inclusion of 0%, 2.5% and 5% FRS. In this period, no significant difference of piglet performance was found among the three groups. From a subset of piglets (n = 10 from each treatment), blood samples for hematology, biochemistry and immunoglobulin analysis, colon digesta for microbiome analysis, and jejunum and colon tissues for histopathological analyses were collected. The piglets fed with 2.5% FRS manifested alleviated intraepithelial and stromal lymphocytes infiltration in the gut, enhanced colon mucosa barrier relative to the 0% FRS group. The colon microbiota composition was determined using V3 and V1-V8 region 16S rRNA gene amplicon sequencing by Illumina NextSeq and Oxford Nanopore MinION, respectively. The two amplicon sequencing strategies showed high consistency between the detected bacteria. Both sequencing strategies indicated that inclusion of FRS reshaped the colon microbiome of weaned piglets with increased Shannon diversity. Prevotella stercorea was verified by both methods to be more abundant in the piglets supplied with FRS feed, and its abundance was positively correlated with colonic mucosa thickness but negatively correlated with blood concentrations of leucocytes and IgG. CONCLUSIONS: FRS supplementation relieved the gut lymphocyte infiltration of the weaned piglets, improved the colon mucosa barrier with altered microbiota composition. Increasing the dietary inclusion of FRS from 2.5% to 5% did not lead to further improvements.

Transcriptomics analysis of differentially expressed genes in subcutaneous and perirenal adipose tissue of sheep as affected by their pre- and early postnatal malnutrition histories.

BACKGROUND: Early life malnutrition is known to target adipose tissue with varying impact depending on timing of the insult. This study aimed to identify differentially expressed genes in subcutaneous (SUB) and perirenal (PER) adipose tissue of 2.5-years old sheep to elucidate the biology underlying differential impacts of late gestation versus early postnatal malnutrition on functional development of adipose tissues. Adipose tissues were obtained from 37 adult sheep born as twins to dams fed either NORM (fulfilling energy and protein requirements), LOW (50% of NORM) or HIGH (110% of protein and 150% of energy requirements) diets in the last 6-weeks of gestation. From day 3 to 6 months of age, lambs were fed high-carbohydrate-high-fat (HCHF) or moderate low-fat (CONV) diets, and thereafter the same moderate low-fat diet. RESULTS: The gene expression profile of SUB in the adult sheep was not affected by the pre- or early postnatal nutrition history. In PER, 993 and 186 differentially expressed genes (DEGs) were identified in LOW versus HIGH and NORM, respectively, but no DEG was found between HIGH and NORM. DEGs identified in the mismatched pre- and postnatal nutrition groups LOW-HCHF (101) and HIGH-HCHF (192) were largely downregulated compared to NORM-CONV. Out of 831 DEGs, 595 and 236 were up- and downregulated in HCHF versus CONV, respectively. The functional enrichment analyses revealed that transmembrane (ion) transport activities, motor activities related to cytoskeletal and spermatozoa function (microtubules and the cytoskeletal motor protein, dynein), and responsiveness to the (micro) environmental extracellular conditions, including endocrine and nervous stimuli were enriched in the DEGs of LOW versus HIGH and NORM. We confirmed that mismatched pre- and postnatal feeding was associated with long-term programming of adipose tissue remodeling and immunity-related pathways. In agreement with phenotypic measurements, early postnatal HCHF feeding targeted pathways involved in kidney cell differentiation, and mismatched LOW-HCHF sheep had specific impairments in cholesterol metabolism pathways. CONCLUSIONS: Both pre- and postnatal malnutrition differentially programmed (patho-) physiological pathways with implications for adipose functional development associated with metabolic dysfunctions, and PER was a major target.

Targeted and Untargeted Metabolic Profiling to Discover Bioactive Compounds in Seaweeds and Hemp Using Gas and Liquid Chromatography-Mass Spectrometry.

Greenhouse gas emissions are a global problem facing the dairy/beef industry. Novel feed additives consisting of seaweeds and hemp containing bioactive compounds are theorized to reduce enteric methane emissions. In this study we aimed to investigate the metabolic profiles of brown, red and green seaweeds and hemp using gas chromatography and liquid chromatography mass spectrometry. We used targeted and untargeted approaches, quantifying known halomethanes and phenolics, as well as identifying potentially novel bioactive compounds with anti-methanogenic properties. The main findings were: (a) Asparagopsis taxiformis contained halomethanes, with high concentrations of bromoform (4200 µg/g DW), six volatile halocarbons were tentatively identified; (b) no halomethanes were detected in the other studied seaweeds nor in hemp; (c) high concentrations of lignans were measured in hemp; (d) a high numbers of sulfated phenolic acids and unidentified sulfuric acid-containing compounds were detected in all seaweeds; (e) flavonoid glucosides and glucuronides were mainly identified in hemp; and (f) the condensed tannin gallocatechin was tentatively identified in Fucus sp. Using the combined metabolomics approach, an overview and in-depth information on secondary metabolites were provided. Halomethanes of Asparagopsis sp. have already been shown to be anti-methanogenic; however, metabolic profiles of seaweeds such as Dictyota and Sargassum have also been shown to contain compounds that may have anti-methanogenic potential.

The influence of feed ingredients on CP and starch disappearance rate in complex diets for broiler chickens.

The influence of feed ingredients on digestion kinetics of N and starch in complex diets was investigated in the current experiment. A total of 34 diets with different inclusion levels of 10 commonly used feed ingredients (corn, wheat, sorghum, soybean meal, canola meal, full-fat soybean meal [FFSB], palm kernel meal, meat and bone meal, wheat distillers grain with solubles and wheat bran) were randomly allocated to 170 cages with 8 birds in each. Apparent jejunal and ileal digestibility of N and starch was determined on a cage level in broilers feed the experimental diets ad libitum from 21 to 24 d after hatch. Disappearance rate of N and starch from the intestine was estimated through a first-order decay function fitted to the digesta data from the jejunum and ileum. The fit of the decay functions was evaluated with root mean squared error as percentage of the observed mean. The influence of the feed ingredients on the disappearance rates were found through a linear regression model, including the effect of the single ingredients, 2-way and 3-way interactions and evaluated with a Student t test test. Starch digestion kinetics were in general faster than N digestion kinetics. The N disappearance rate was both influenced by single ingredients and interaction amongst ingredients, whereas starch disappearance rate mainly was influenced by single ingredients. A combination of FFSB and soybean meal decreased the N digestion rate by 22 to 25% compared with diets with only soybean meal or FFSB, respectively. These results indicate that nutrients from 1 feed ingredient can influence the rate of disappearance of nutrients from other feed ingredients in a complex diet. This highlights the importance of understanding nutrient digestion kinetics and how these are influenced both additively and nonadditively by different feed ingredients in complex diets.

Depot and sex-specific implications for adipose tissue expandability and functional traits in adulthood of late prenatal and early postnatal malnutrition in a precocial sheep model.

The aim was to investigate long-term, tissue and sex-specific impacts of pre and postnatal malnutrition on expandability and functional traits of different adipose tissues. Twin-pregnant ewes were fed NORM (~requirements), LOW (50% of NORM) or HIGH (150%/110% of energy/protein) diets the last 6 weeks prepartum (term ~147-days). Lambs received moderate, low-fat (CONV) or high-carbohydrate-high-fat (HCHF) diets from 3 days until 6 months of age, and thereafter CONV diet. At 2½ years of age (adulthood), histomorphometric and gene expression patterns were characterized in subcutaneous (SUB), perirenal (PER), mesenteric (MES), and epicardial (EPI) adipose tissues. SUB had sex-specific (♂<♀) upper-limits for adipocyte size and cell-number indices, irrespective of early life nutrition. PER mass and contents of adipocytes were highest in females and HIGH♂, whereas adipocyte cross-sectional area was lowest in LOW♂. Pre/postnatal nutrition affected gene expression sex-specifically in SUB + PER, but unrelated to morphological changes. In PER, LOW/LOW♂ were specific targets of gene expression changes. EPI was affected by postnatal nutrition, and HCHF sheep had enlarged adipocytes and upregulated expressions for adipogenic and lipogenic genes. Conclusion: upper-limits for SUB expandability were markedly lower in males. Major targets for prenatal malnutrition were PER and males. LOW♂ had the lowest PER expandability, whereas HIGH♂ had an adaptive advantage due to increased hypertrophic ability equivalent to females. Fixed expandability in SUB meant PER became a determining factor for MES and ectopic fat deposition, rendering LOW♂ particularly predisposed for obesity-associated metabolic risks. EPI, in contrast to other tissues, was targeted particularly by early postnatal obesity, resulting in adipocyte hypertrophy in adulthood.

Effects of Increasing Doses of Lactobacillus Pre-Fermented Rapeseed Product with or without Inclusion of Macroalgae Product on Weaner Piglet Performance and Intestinal Development.

This study evaluated the effects of increasing doses of pre-fermented rapeseed meal (FRM) without or with inclusion of the brown macroalgae Ascophyllum nodosum (AN) on weaner piglets' performance and gut development. Ten days pre-weaning, standardized litters were randomly assigned to one of nine isoenergetic and isoproteic diets comprising (on DM basis): no supplement (negative control, NC), 2500 ppm ZnO (positive control, PC), 8, 10, 12, 15 or 25% FRM, and 10% FRM plus 0.6 or 1.0% AN. Fifty piglets receiving the same pre-weaning diets were weaned at 28 days of age and transferred to one pen, where they continued on the pre-weaning diet until day 92. At 41 days, six piglets per treatment were sacrificed for blood and intestinal samplings. The average daily gain was at least sustained at any dose of FRM (increased at 8% FRM, 28-41 days) from 18-41 days similar to PC but unaffected by inclusion of AN. The percentage of piglets that completed the experiment was increased by FRM compared to NC, despite detection of diarrhea symptoms. FRM showed quadratic dose-response effects on colon and mid-jejunum crypts depth, and enterocyte and mid-jejunum villus heights with optimum development at 8% or 10% FRM, respectively, but this was abolished when AN was also added. In conclusion, FRM sustained piglet growth performance and intestinal development similar to ZnO with an optimum inclusion level of 8-10% of dietary DM.

The Effect of Feeding Roughages of Varying Digestibility Prepartum on Energy Status and Metabolic Profiles in Beef Cows around Parturition.

Resource efficient winter-feeding of mature pregnant beef cows requires knowledge of how different roughage-based feeding strategies affect cow intake and energy status. Four diets based on traditional timothy-meadow fescue silage (TM), festulolium silage plus urea (FE), reed canarygrass silage (RC) or barley straw supplemented with urea and rapeseed meal (BR), were fed ad libitum for 16 weeks prepartum to 36 Hereford cows. Postpartum, cows were fed the same diet before release on pasture. Individual data on cow intake, changes in body weight (BW), body condition score (BCS) and plasma metabolites, calf birth and weaning weights were recorded. The TM and FE diets resulted in increased BW and BCS prepartum (p < 0.001), while the RC and BR diets resulted in a catabolic state, as indicated by a loss of BCS, lower insulin levels and higher non-esterified fatty acid levels in cows fed BR (p < 0.001). There were no dietary effects on calf parameters (p > 0.29). Feeding RC or BR prepartum might be a possible alternative to traditional timothy-meadow fescue silage if cows are allowed to regain lost BCS during the grazing period. The influence on cow reproductive- and calf performance should be considered before making this management change.

Is Foetal Programming by Mismatched Pre- and Postnatal Nutrition Contributing to the Prevalence of Obesity in Nepal?

Nepal and many developing countries are currently suffering from increased prevalence of obesity, type 2 diabetes, and other metabolic disorders. Unhealthy dietary habits and physical inactivity are traditionally considered as responsible factors for these disorders. The relatively new concept of foetal programming suggests that development of metabolic diseases later in life may be associated with poor nutritional status in utero, and such phenomenon could be amplified by subsequent exposure to unhealthy diets after birth. We suggest that foetal programming and mismatched nutritional situations during foetal and postnatal life are important causative factors for increased prevalence of obesity and metabolic disorders in Nepal. Issues highlighted in this paper may also be relevant to other developing countries with similar socioeconomic status. Undernutrition in foetal life can predispose for visceral fat deposition and may alter dietary preferences towards unhealthy diets, amplifying the risk of nutritional mismatch after birth; this can lead to metabolic disturbances in a number of pathways including glucose and lipid metabolism. Providing attention to early life nutrition could therefore be an important tool to reduce the prevalence of lifestyle diseases in Nepal. Future national health policies should thus include changes in research and intervention activities towards preventing averse early life nutritional programming. Availability of free-of-cost and mandatory nutritional education and medical services to pregnant women and their families and better management of national health care systems including digitalization of national health data could be viable strategies to achieve these goals.

Hypothalamic oxidative stress and inflammation, and peripheral glucose homeostasis in Sprague-Dawley rat offspring exposed to maternal and postnatal chocolate and soft drink.

BACKGROUND: Predisposition to obesity and type 2 diabetes can arise during foetal development and in early postnatal life caused by imbalances in maternal nutritional overload. We aimed to investigate the effects of maternal and postnatal intake of chocolate and soft drink on hypothalamic anti-oxidative stress markers, inflammation and peripheral glucose homeostasis. METHODS: Pregnant Sprague-Dawley rats were fed ad libitum chow diet only (C) or with chocolate and high sucrose soft drink supplements (S). At birth, litter size was adjusted into 10 male offspring per dam. After weaning at 3 weeks of age, offspring from both dietary groups were assigned to either S or C diet, giving four groups until the end of the experiment at 26 weeks of age. RESULTS: Offspring exposed to maternal S had up-regulated hypothalamic anti-oxidative markers such as SOD2 and catalase at 3 weeks of age as an indication of oxidative stress. However, at 12 weeks of age these anti-oxidative markers tended to decrease while pro-inflammatory markers such as TNF and IL-1ß became up-regulated of all offspring exposed to S diet during some point of their life. Thus, despite an increase in anti-oxidative stress response, offspring exposed to maternal S had a reduced ability to counteract hypothalamic inflammation. At the same time point, postnatal S resulted in increased adiposity, reduced glucose tolerance and insulin sensitivity with no effect on body weight. However, at 25 weeks of age, the impaired glucose tolerance was reversible to the response of the control regardless of increased adiposity and body weight pointing towards a compensatory response of the insulin sensitivity or insulin secretion. CONCLUSION: Indications of hypothalamic oxidative stress was observed prior to the inflammatory response in offspring exposed to maternal S. Both maternal and postnatal S induced hypothalamic inflammation prior to increased weight gain and thus contributing to obese phenotype.

Impacts of pre- and postnatal nutrition on glucagon regulation and hepatic signalling in sheep.

To evaluate the long-term impacts of early-life nutritional manipulations on glucagon secretion and hepatic signalling, thirty-six twin-pregnant ewes during their last trimester were exposed to NORM (fulfilling 100% of daily energy/protein requirements), HIGH (fulfilling 150/110% of daily energy/protein requirements) or LOW (50% of NORM) diets. Twin lambs were assigned after birth to a moderate (CONV) or high-carbohydrate high-fat (HCHF) diet until 6 months. Then, responses in plasma glucagon concentrations and glucagon ratios relative to previously reported values for insulin, glucose and lactate were determined after intravenous bolus injections of glucose or propionate (fed and 2-day fasting state). Hepatic mRNA expressions of glucagon receptor (GCGR), glucose-6-phosphatase (G6PC), phosphoenolpyruvate carboxykinase (PEPCK) and fructose 1,6-biphosphatase (FBP) were also determined in a sub group of autopsied lambs. Expression of GCGR and all three enzymes were supressed by prenatal LOW compared to NORM (except PEPCK) and HIGH (except FBP) nutrition. The postnatal HCHF diet reduced plasma glucagon responses to propionate and hepatic mRNA expression of all genes. In response to propionate, insulin/glucagon ratio was decreased (fasted state), but lactate/glucagon and glucose/glucagon increased in HCHF compared to CONV lambs. In conclusion, prenatal undernutrition and postnatal overnutrition had similar long-term implications and reduced hepatic glucagon signalling. Glucagon secretory responses to propionate were, however, not related to the prenatal nutrition history, but negatively affected by the postnatal obesogenic diet. The pancreatic α-cell compared to ß-cells may thus be less sensitive towards late gestation malnutrition, whereas hepatic glucagon signalling appears to be a target of prenatal programming.

Impacts of prenatal nutrition on animal production and performance: a focus on growth and metabolic and endocrine function in sheep.

The concept of foetal programming (FP) originated from human epidemiological studies, where foetal life nutrition was linked to health and disease status later in life. Since the proposal of this phenomenon, it has been evaluated in various animal models to gain further insights into the mechanisms underlying the foetal origins of health and disease in humans. In FP research, the sheep has been quite extensively used as a model for humans. In this paper we will review findings mainly from our Copenhagen sheep model, on the implications of late gestation malnutrition for growth, development, and metabolic and endocrine functions later in life, and discuss how these implications may depend on the diet fed to the animal in early postnatal life. Our results have indicated that negative implications of foetal malnutrition, both as a result of overnutrition and, particularly, late gestation undernutrition, can impair a wide range of endocrine functions regulating growth and presumably also reproductive traits. These implications are not readily observable early in postnatal life, but are increasingly manifested as the animal approaches adulthood. No intervention or cure is known that can reverse this programming in postnatal life. Our findings suggest that close to normal growth and slaughter results can be obtained at least until puberty in animals which have undergone adverse programming in foetal life, but manifestation of programming effects becomes increasingly evident in adult animals. Due to the risk of transfer of the adverse programming effects to future generations, it is therefore recommended that animals that are suspected to have undergone adverse FP are not used for reproduction. Unfortunately, no reliable biomarkers have as yet been identified that allow accurate identification of adversely programmed offspring at birth, except for very low or high birth weights, and, in pigs, characteristic changes in head shape (dolphin head). Future efforts should be therefore dedicated to identify reliable biomarkers and evaluate their effectiveness for alleviation/reversal of the adverse programming in postnatal life. Our sheep studies have shown that the adverse impacts of an extreme, high-fat diet in early postnatal life, but not prenatal undernutrition, can be largely reversed by dietary correction later in life. Thus, birth (at term) appears to be a critical set point for permanent programming in animals born precocial, such as sheep. Appropriate attention to the nutrition of the late pregnant dam should therefore be a priority in animal production systems.

Long-Term Impacts of Foetal Malnutrition Followed by Early Postnatal Obesity on Fat Distribution Pattern and Metabolic Adaptability in Adult Sheep.

We aimed to investigate whether over- versus undernutrition in late foetal life combined with obesity development in early postnatal life have differential implications for fat distribution and metabolic adaptability in adulthood. Twin-pregnant ewes were fed NORM (100% of daily energy and protein requirements), LOW (50% of NORM) or HIGH (150%/110% of energy/protein requirements) diets during the last trimester. Postnatally, twin-lambs received obesogenic (HCHF) or moderate (CONV) diets until 6 months of age, and a moderate (obesity correcting) diet thereafter. At 2½ years of age (adulthood), plasma metabolite profiles during fasting, glucose, insulin and propionate (in fed and fasted states) tolerance tests were examined. Organ weights were determined at autopsy. Early obesity development was associated with lack of expansion of perirenal, but not other adipose tissues from adolescence to adulthood, resulting in 10% unit increased proportion of mesenteric of intra-abdominal fat. Prenatal undernutrition had a similar but much less pronounced effect. Across tolerance tests, LOW-HCHF sheep had highest plasma levels of cholesterol, urea-nitrogen, creatinine, and lactate. Sex specific differences were observed, particularly with respect to fat deposition, but direction of responses to early nutrition impacts were similar. However, prenatal undernutrition induced greater metabolic alterations in adult females than males. Foetal undernutrition, but not overnutrition, predisposed for adult hypercholesterolaemia, hyperureaemia, hypercreatinaemia and hyperlactataemia, which became manifested only in combination with early obesity development. Perirenal expandability may play a special role in this context. Differential nutrition recommendations may be advisable for individuals with low versus high birth weights.