Distribution of ghrelin-producing cells in the gastrointestinal tract of pigs at different ages

Ghrelin is involved in many biological processes, ranging from appetite regulation and the release of growth hormone to the regulation of gastrointestinal motility and secretion processes. Ghrelin expression is not homogenously distributed throughout the gastrointestinal tract; expression is species-specific and can also depend on the animal age. This study was performed to investigate ghrelin immunolocalization in the gastrointestinal tract of pigs at different ages: 1 day (birth), 28 days (weaning), 2 months, 4 months, and 7 months (pre-puberty). Tissue samples were collected along the entire gastrointestinal tract and were examined by immunohistochemistry and double-immunofluorescence. Histometry was performed by counting the number of endocrine ghrelin immunopositive cells in the gastrointestinal mucosa. Ghrelin was found to be present along the swine alimentary canal from the stomach to the caecum. In all regions of the alimentary canal of the animals studied, ghrelin-immunoreactive (IR) cells co-localized with chromogranin-A and were therefore identified as endocrine cells. In the gastric fundus, ghrelin-immunoreactivity was partially detected in co-localization with H-K-adenosine triphosphatase and pepsinogen. Ghrelin-IR endocrine cells were abundant in the oxyntic mucosa but less present in the small intestine and rare in the large intestine. The cell density of the ghrelin-IR endocrine cells was lowest in the oxyntic mucosa of 1-day-old pigs. We can conclude that gastric ghrelin expression is not related merely to age but could also potentially be influenced by food intake.     

Effects of Pre‐Weaning Energy Substitutions on Post‐Weaning Follicle Development,Steroid Hormones and Subsequent Litter Size in Primiparous Sows

The present study investigated the effects of pre‐weaning energy substitutions on follicular development, endocrine characteristics and subsequent litter size in primiparous sows. Sows were fed a standard lactation diet (14.1 DE MJ/kg) and then allocated to a Control (C, n = 24), Fat (F, n = 23), Sugar (S, n = 23) or post‐weaning Regumate (positive control; R, n = 22) treatment at 9 days before weaning of the C, F and S treatments. During the treatment period (8 days), 1 kg of the lactation diet was substituted with 1 kg of a fat‐rich (F, 23.85 DE MJ/kg) or sugar‐rich (S, 15.75 DE MJ/kg) substitution for F and S sows, respectively. For the R treatment, sows were weaned 8 days earlier than other treatments and fed a lactation diet at 3.5 kg with two doses of altrenogest as topdressing from 1 day before weaning until the day on which the other sows were weaned. The F treatment aimed to increase energy intake, and the S treatment aimed to elevate post‐prandial glucose and insulin concentrations. Weaning‐to‐ovulation interval tended to be reduced in the S treatment compared with C (p = 0.06) and F (p = 0.08) treatments. Body weight (BW) loss during the treatment period, post‐weaning follicle development, plasma oestradiol and pre‐weaning leptin did not differ among C, F and S sows, although BW loss was lower and leptin was higher in the R treatment. Post‐ovulatory progesterone concentration in the S treatment was higher (p < 0.05). Sows in the S and R treatments had a greater proportion of litters with larger litter sizes (p < 0.05). The outcome suggests that increasing circulating insulin and glucose concentrations during late lactation or a week of metabolic recovery positively improves subsequent litter size in primiparous sows.     

Effects of Dietary Supplementation of β‐hydroxy‐β‐methylbutyrate on Sow Performance and mRNA Expression of Myogenic Markers in Skeletal Muscle of Neonatal Piglets

The effects of dietary β‐hydroxy‐β‐methylbutyrate (HMB) supplementation during gestation on reproductive performance of sows and the mRNA expression of myogenic markers in skeletal muscle of neonatal pigs were determined. At day 35 of gestation, a total of 20 sows (Landrace × Yorkshire, at third parity) were randomly assigned to two groups, with each group receiving either a basal diet or the same diet supplemented with 4 g/day β‐hydroxy‐β‐methylbutyrate calcium (HMB‐Ca) until parturition. At parturition, the total and live litter size were not markedly different between treatments, however, the sows fed HMB diet had a decreased rate of stillborn piglets compared with the sows fed the control (CON) diets (p < 0.05). In addition, piglets from the sows fed HMB diet tended to have an increased birth weight (p = 0.08), and a reduced rate of low birth weight piglets (p = 0.05) compared with piglets from the CON sows. Nevertheless, lower feed intake during lactation was observed in the sows fed the HMB diet compared with those on the CON diet (p < 0.01). The relative weights of the longissimus dorsi (LD) and semitendinosus (ST) muscle were higher (p < 0.05) in neonatal pigs from the HMB than the CON sows. Furthermore, maternal HMB treatment increased the mRNA levels of the myogenic genes, including muscle regulatory factor‐4 (MRF4, p < 0.05), myogenic differentiation factor (MyoD) and insulin‐like growth factor‐1 (IGF‐1, p < 0.01). In conclusion, dietary HMB supplementation to sows at 4 g/day from day 35 of gestation to term significantly improves pregnancy outcomes and increases the expression of myogenic genes in skeletal muscle of neonatal piglets, but reduces feed intake of sows during lactation.     

Carcass characteristics,chemical and fatty acid composition and oxidative stability of meat from broiler chickens fed black cumin (Nigella sativa) seeds

A study was conducted to investigate the dietary supplementation of black cumin seeds (BCS) on carcass characteristics, chemical and fatty acid (FA) composition and antioxidant properties of thigh and breast meat of broiler chickens at 42 days of age. Three hundred sixty 1‐day‐old broiler chickens were allocated to five dietary treatment groups (each group containing eight replicate pens with each pen containing nine broiler chickens): basal diet (control; CON), CON + 0.05 g/kg of bacitracin methylene disalicylate (AB), CON + 5 g/kg of BCS (low dose of BCS), CON + 10 g/kg of BCS (medium dose of BCS) and CON + 20 g/kg of BCS (high dose of BCS). Weight (g) of slaughtered birds (p = .03), hot carcass (p = .007), breast (p = .03), thigh (p < .001), wing (p = .06), neck (p = .01), liver (p = .09), abdominal fat (p = .01) and total edible parts (p = .01) increased or tended to increase due to BCS supplementation compared with the CON. The concentrations of dry matter, crude protein and ether extract in chicken thigh and breast meat increased (p = .038 to <.001) with increasing doses of BCS in diets. The ferric reducing antioxidant activity in blood and meat increased linearly with increasing doses of BCS in the diets. However, peroxide values in meat were not affected by BCS and AB on both days 1 and 7 of storage at 4°C. Supplementation of BCS increased the concentrations of C14:1, C18:3n‐6, C20:1, C20:2 FA and PUFA linearly (p < .05) and tended to increase (p = .098) the concentration of C18:2cis linearly. However, the concentrations of C16:0 and C16:1 FA decreased linearly with increasing doses of BCS in the diets. In conclusion, dietary supplementation of BCS at 20 g/kg diet may improve slaughter body weight, beneficial FA concentrations and antioxidant properties of broiler chicken meat.     

Flutamide Influences Placental Aldo–Keto Reductase Family 1 Member C1 (AKR1C1) Expression in Pigs

Aldo–keto reductase family 1 member C1 (AKR1C1) catalyses the conversion of progesterone into inactive 20α‐dihydroxyprogesterone. It is suggested that AKR1C1 expression in the placenta prevents from the cytotoxic effect of progesterone on foetuses during late pregnancy. The aim of the study was to determine whether the anti‐androgen flutamide administered during late pregnancy (83–89 days of gestation) or before parturition (101–107 days of gestation) influences AKR1C1 expression in the porcine placenta. AKR1C1 mRNA and protein levels were measured using real‐time PCR and western blotting, respectively. Immunolocalization of AKR1C1 within placentas was also performed. Flutamide significantly increased AKR1C1 mRNA (p = 0.008) and protein (p = 0.019) expression only during the pre‐parturient period in pigs. AKR1C1 protein was immunolocalized in the epithelial and stromal cells of foetal and maternal part of placenta at both stages of gestation. Following flutamide treatment, the intensity of staining was higher (p = 0.045) on day 108 of gestation. In conclusion, porcine placental AKR1C1 expression seems to be regulated by an androgen signalling pathway and may be involved in foetal survival by preventing the detrimental effect of progesterone.     

Dietary levels of protein and free amino acids affect pancreatic proteases activities,amino acids transporters expression and serum amino acid concentrations in starter pigs

The dietary contents of crude protein and free amino acids (AA) may affect the protein digestion and AA absorption in pigs. Trypsin and chymotrypsin activities, AA serum concentrations and expression of AA transporters in the small intestine of pigs fed a low protein, AA‐supplemented (19.2%, LPAA) or a high protein (28.1%, HP), wheat‐soybean meal diet were measured in two 14‐d trials. The LPAA diet contained free L‐Lys, L‐Thr, DL‐Met, L‐Leu, L‐Ile, L‐Val, L‐His, L‐Trp and L‐Phe. All pigs were fed the same amount of feed (890 and 800 g/d for trial 1 and 2 respectively). In trial 1, samples of mucosa (duodenum, jejunum and ileum) and digesta (duodenum and jejunum) were collected from 14 pigs (17.2 ± 0.4 kg); in trial 2, blood samples were collected from 12 pigs (12.7 ± 0.3 kg). The trypsin and chymotrypsin activities in both intestinal segments were higher in pigs fed the HP diet (p < 0.01). Trypsin activity was higher in jejunum than in duodenum regardless the dietary treatment (p < 0.05). Pigs fed the LPAA diet expressed more b^0,+AT in duodenum, B⁰AT1 in ileum (p < 0.05), and tended to express more y⁺LAT1 in duodenum (p = 0.10). In pigs fed the LPAA diet, the expression of b^0,+AT was higher in duodenum than in jejunum and ileum (p < 0.01), but no difference was observed in pigs fed the HP diet. Ileum had the lowest b^0,+AT expression regardless the diet. The serum concentrations of Lys, Thr and Met were higher in LPAA pigs while serum Arg was higher in HP pigs (p < 0.05). Serum concentrations of AA appear to reflect the AA absorption. In conclusion, these data indicate that the dietary protein contents affect the extent of protein digestion and that supplemental free AA may influence the intestinal site of AA release and absorption, which may impact their availability for growth of young pigs.     

Essential oils improved weight gain,growth and feed efficiency of young dairy calves fed 18 or 20% crude protein starter diets

The objective was to evaluate interactions between starter protein (180 vs. 200 g/kg, DM basis) and a mixture of essential oils (EOs; containing thymol, eugenol, vanillin, limonene and guaiacol) on growth, metabolic and ruminal functions of Holstein dairy calves. In a completely randomized 2 × 2 factorial design, 48 calves, 3 days old (averaging BW 42.7 ± 1.9 kg), were allocated into groups fed the following diets: (i) 180 g/kg CP with no EO (180P‐NEO); (ii) 180 g/kg CP with EO (180P‐EO); (iii) 200 g/kg CP with no EO (200P‐NEO); and (iv) 200 g/kg CP with EO (200P‐EO). The EO was supplemented as 1 g/kg of starter DM. Calves were fed ad libitum starter diet and were weaned at day 59 of age, but diets continued until day 80. There were no interactive effects of CP and EO on intake and growth. Pre‐weaning feed efficiency tended to be increased for 200P‐EO (p = .09). Average daily gain and feed efficiency during pre‐weaning period as well as weaning weight were increased (p < .05) by EO, whereas wither height was increased by EO (p = .03) and tended to be increased for 200P vs. 180P (p = .06). Post‐weaning blood urea nitrogen concentration tended to be lower in 180P vs. 200P (p = .08). Ruminal short‐chain fatty acids concentration was greatest in 200P‐EO. The EO increased both butyrate (p = .02) and propionate proportions (p = .01) and reduced acetate proportional ratio (p < .01). Ruminal ammonia‐N was tended to be lower in calves‐fed EO (p = .05) and was lower in those fed 180P vs. 200P (p < .01). In conclusion, supplementation of the starter diet with essential oil improved weight gain, growth and feed efficiency of dairy calves, irrespective of dietary protein content.     

Effect of dietary fish oil on fatty acid deposition and expression of cholesterol homeostasis controlling genes in the liver and plasma lipid profile: comparison of two animal models

The objective of the present study was to compare hepatic fatty acid deposition, plasma lipid level and expression of cholesterol homeostasis controlling genes in the liver of rats (Wistar Albino; n = 32) and pigs (Large White × Landrace; n = 32) randomly assigned into two groups of 16 animals each and fed 10 weeks the diet with either 2.5% of fish oil (F; source of eicosapentaenoic and docosahexaenoic acid, EPA+DHA) or 2.5% of palm oil (P; high content of saturated fatty acids; control). F‐rats deposited in the liver three times less EPA, but 1.3 times more DHA than F‐pigs (p < 0.05). Dietary fish oil relative to palm oil increased PPARα and SREBP‐2 gene expression much strongly (p < 0.01) in the pig liver in comparison with the rat liver, but expression of Insig‐1 and Hmgcr genes in the liver of the F‐pigs relative to the expression of these genes in the liver of the P‐pigs was substantially lower (p < 0.01 and p < 0.05 respectively) as compared to rats. When plasma lipid concentration in the F‐animals was expressed as a ratio of the plasma concentration in the P‐counterparts, dietary fish oil decreased HDL cholesterol less (p < 0.01), but LDL cholesterol and triacylglycerols more (p < 0.05 and p < 0.001 respectively) in rats than in pigs: more favourable effect of fish oil on rat plasma lipids in comparison with pigs can therefore be concluded. Concentration of total cholesterol and both its fractions in the rat plasma was negatively correlated (p < 0.01) with hepatic DHA, but also with unsaturated myristic and palmitic acid respectively. It has been concluded that regarding the similarity of the plasma lipid levels to humans, porcine model can be considered superior; however, using this model, dietary fish oil at the tested amount (2.5%) was not able to improve plasma lipid markers in comparison with saturated palm oil.     

Differential Ovarian Morphometry and Follicular Expression of BMP15, GDF9 and BMPR1B Influence the Prolificacy in Goat

The variation in the kidding size of Black Bengal and Sirohi breed of goats makes them an interesting genetic material to study the underlying genetic mechanism of prolificacy. Accordingly, we studied the comparative ovarian morphometry including disparity in numbers of antral follicles of different sizes between these two breeds. Further, we evaluated the differential expression of the important candidate genes (viz., BMP15, GDF9 and BMPR1B) known to influence the ovulation rates and the prolificacy. The ovaries of Black Bengal (n = 20) goat were lighter (p < 0.01) in weight and smaller (p < 0.01) in diameter than those of Sirohi (n = 19) goats but possessed more numbers (p < 0.05) of corpus luteum (CL), large and small antral follicles. Quantitative real‐time PCR (RT‐qPCR) analysis revealed differential expression of mRNAs encoding for the BMP15 and GDF9. Small antral follicles of Black Bengal goats expressed 2.78‐fold more (p < 0.05) of BMP 15 than those of Sirohi goat. Expression of BMP15 (p < 0.01) and GDF9 (p < 0.05) mRNAs was more abundant in the small than the large antral follicles of Black Bengal goat. The more numbers of antral follicles per unit of ovarian mass and differential expression of BMP15 and GDF9 may serve as an important clue for higher prolificacy.     

Comparison of Diverse Differential Plating Methods to Enrich Bovine Spermatogonial Cells

Spermatogonial stem cells (SSC) have important applications in domestic animal reproduction and advanced biotechnologies. Because differential plating is one of the most common methods used for SSC enrichment, the goal of this study was to compare three differential plating methods for the enrichment of bovine SSC. To achieve this goal, testicular parenchyma from pre‐pubertal calves was minced and single cells were obtained after two enzymatic digestions. We compared three coating methods for differential plating: laminin (20 ng/ml), BSA (0.05 mg/ml) and PBS. Cells were incubated at 37°C, 5% CO₂ in air for 15 min onto laminin‐coated dishes or 2 h onto BSA‐ or PBS‐coated dishes. Cell viability was assessed by trypan blue exclusion method. Recovered cells were analysed for the expression of SSC molecular markers by quantitative RT‐PCR (GFRA1, CXCR4, ITGA6, THY1) and flow cytometry (GFRA1, CXCR4 and ITGA6). Cells at time 0, adherent cells on laminin and non‐adherent cells from BSA and PBS groups had the same cell viability (p = 0.0655). GFRA1, CXCR4 and THY1 relative gene expression was higher (p = 0.0402, p = 0.0007, p = 0.0117, respectively) for non‐adherent cells selected in PBS group. Flow cytometry analysis revealed that the presence of GFRA‐positive (GFRA+) cells was higher in non‐adherent cells from BSA and PBS groups (p < 0.001). However, laminin‐adherent cells had higher number of ITGA6+ cells (p < 0.001) and lower presence of CXCR4+ cells (p = 0.0012). In conclusion, differential plating is an effective method for the enrichment of bovine undifferentiated spermatogonia and higher expression of SSC markers is obtained without laminin or BSA coating.     

Ghrelin Accelerates In Vitro Maturation of Bovine Oocytes

Ghrelin, apart from its metabolic role, is nowadays considered as a basic regulator of reproductive functions of mammals, acting at central and gonadal levels. Here, we investigated for possible direct actions of ghrelin on in vitro maturation of bovine oocytes and for its effects on blastocyst yield and quality. In experiment 1, cumulus oocyte complexes (COCs) were matured in the presence of four different concentrations of ghrelin (0, 200, 800 and 2000 pg/ml). In vitro fertilization and embryo culture were carried out in the absence of ghrelin, and blastocyst formation rates were examined on days 7, 8 and 9. In experiment 2, only the 800 pg/ml dose of ghrelin was used. Four groups of COCs were matured for 18 or 24 h (C18, Ghr18, C24 and Ghr24), and subsequently, they were examined for oocyte nuclear maturation and cumulus layer expansion; blastocysts were produced as in experiment 1. The relative mRNA abundance of various genes related to metabolism, oxidation, developmental competence and apoptosis was examined in snap‐frozen cumulus cells, oocytes and day‐7 blastocysts. In experiment 1, ghrelin significantly suppressed blastocyst formation rates. In experiment 2, more ghrelin‐treated oocytes matured for 18 h reached MII compared with controls, while no difference was observed when maturation lasted for 24 h. At 18 and 24 h, the cumulus layer was more expanded in ghrelin‐treated COCs than in the controls. The blastocyst formation rate was higher in Ghr18 (27.7 ± 2.4%) compared with Ghr24 (17.5 ± 2.4%). Differences were detected in various genes’ expression, indicating that in the presence of ghrelin, incubation of COCs for 24 h caused over‐maturation (induced ageing) of oocytes, but formed blastocysts had a higher hatching rate compared with the controls. We infer that ghrelin exerts a specific and direct role on the oocyte, accelerating its maturational process.     

Paraoxonase (PON) 1, 2 and 3 Expression in Granulosa Cells and PON1 Activity in Follicular Fluid of Dairy Cows

Normal metabolic activity in ovarian follicles may result in oxidative stress and damage to oocytes. The aim of this study was to evaluate expression of the natural anti‐oxidants paraoxonase (PON) 1, 2 and 3 in granulosa cells and PON1 activity in follicular fluid (FF) and plasma of dairy cows. For the first experiment, ovaries were collected from cows at slaughter, after which follicles were dissected and classified as oestrogen active (EAF) or atretic (ATF). Expression of PON1, PON2 and PON3 mRNA was evaluated in granulosa cells, and activity of PON1 was measured in FF. PON1 mRNA was undetectable in granulosa cells, PON2 mRNA expression was not different between follicle types, and PON3 mRNA tended to be higher in EAF (p = 0.11). The activity of PON1 in FF was higher (p = 0.01) for EAF (82.6 ± 8.0 kU/L) than ATF (53.9 ± 6.8 kU/L), as were high‐density lipoproteins (HDL), low‐density lipoproteins (LDL) and total cholesterol concentrations. In the second experiment, we aimed to compare plasma and FF PON1 activity in early lactation Holstein cows (n = 15) with pre‐ovulatory EAF. Activity of PON1 was twofold higher (p < 0.0001) in plasma (122.5 ± 11.1 kU/L) than in FF (61.4 ± 5.2 kU/L). Plasma concentrations were also higher (p < 0.0001) for HDL, LDL and total cholesterol when compared to FF. In conclusion, FF concentrations of PON1, HDL, LDL and total cholesterol were higher in healthy oestrogen active bovine follicles than in atretic follicles. PON1 was not expressed by granulosa cells indicating that high PON1 activity in bovine FF is apparently derived by transfer from blood in association with HDL.     

Effects of supplementation with green tea by‐products on growth performance,meat quality,blood metabolites and immune cell proliferation in goats

Forty‐eight castrated male goats were used to determine the effects of feeding green tea by‐products (GTB) on growth performance, meat quality, blood metabolites and immune cell proliferation. Experimental treatments consisted of basal diets supplemented with four levels of GTB (0%, 0.5%, 1.0% or 2.0%). Four replicate pens were assigned to each treatment with three goats per replicate. Increasing dietary GTB tended to linearly increase the overall average weight gain and feed intake (p = 0.09). Water holding capacity, pH and sensory attributes of meat were not affected by GTB supplementation, while cooking loss was reduced both linearly and quadratically (p < 0.01). The redness (linear; p = 0.02, quadratic; p < 0.01) and yellowness (quadratic; p < 0.01) values of goat meat were improved by GTB supplementation. Increasing dietary GTB quadratically increased protein and decreased crude fat (p < 0.05), while linearly decreased cholesterol (p = 0.03) content of goat meat. The proportions of monounsaturated fatty acid, polyunsaturated fatty acid (PUFA) and n‐6 PUFA increased linearly (p < 0.01) and n‐3 PUFA increased quadratically (p < 0.05) as GTB increased in diets. Increasing dietary GTB linearly increased the PUFA/SFA (saturated fatty acid) and tended to linearly and quadratically increase (p ≤ 0.10) the n‐6/n‐3 ratio. The thiobarbituric acid‐reactive substances values of meat were lower in the 2.0% GTB‐supplemented group in all storage periods (p < 0.05). Dietary GTB linearly decreased plasma glucose and cholesterol (p < 0.01) and quadratically decreased urea nitrogen concentrations (p = 0.001). The growth of spleen cells incubated in concanavalin A and lipopolysaccharides medium increased significantly (p < 0.05) in response to GTB supplementation. Our results suggest that GTB may positively affect the growth performance, meat quality, blood metabolites and immune cell proliferation when supplemented as a feed additive in goat diet.     

Incorporation of sunflower oil or linseed oil in equine compound feedstuff: 1 Effects on haematology and on fatty acids profiles in the red blood cells membranes

Eight trained horses (6 mares – 2 geldings, 6 Selle Français, 2 Trotteur Français, 12 ± 5.8 years old, 538 ± 72.5 kg) were offered three diets to potentially affect haematology and the fatty acids (FA) profiles in red blood cells (RBC) membranes. The control diet was composed of 50% hay and 50% concentrate containing mainly rolled barley (48%) and whole spelt (48%). In the case of sunflower oil diet, sunflower oil (62.0% of α‐linoleic acid, LA) was incorporated at a rate of 8% and substituted by an equal proportion of barley. In the linseed oil diet, first cold‐pressed linseed oil (56.0% of α‐linolenic acid, ALA) was utilised at a similar incorporation rate of 8%. The experimental design consisted of three 3 × 3 latin squares with one being incomplete. Each period lasted 8 weeks. On average, the total feed intake (straw excluded) was 6.2 kg/day and the oil intake 0.278 kg/day. The oils significantly increased the concentrations of RBC, haemoglobin and haematocrit. The oils had no significant impact on the haematology profiles except that platelets tended to decrease in both oil‐based diets. The most abundant FA in the RBC membranes of the control diet samples were in the decreasing order LA, C18:1n9‐7, C18:0, C16:0 and the arachidonic acid (ARA) respectively. The sunflower oil supplementation slightly increased the amount of LA (36.23 vs. 34.72 mg/dl, p = 0.55) and C22:4n‐6 (0.21 vs. 0.09 mg/dl, p = 0.22), while the decrease was observed in case of other FA (C16:1n‐7, 1.08 vs. 1.42 mg/dl, p = 0.03), C20:3n‐6 (0.22 vs. 0.31 mg/dl, p = 0.02), and ARA (1.17 vs. 1.63 mg/dl, p = 0.08). Linseed oil induced similar effects in the n‐6 series FA profiles. In the context of practical applications, our results show that linseed oil incorporation in the diet could improve the haematology and the n‐3 FA profiles potentially leading to an increased performance.     

Methyl donors dietary supplementation to gestating sows diet improves the growth rate of offspring and is associating with changes in expression and DNA methylation of insulin‐like growth factor‐1 gene

The study aimed to investigate the effects of maternal dietary methyl donors on the performance of sows and their offspring, and the associated hepatic insulin‐like growth factor‐1 (IGF‐1) expression of the offspring. A total of 24 multiparous sows were randomly fed the control (CON) or the CON diet supplemented with methyl donors (MD) at 3 g/kg betaine, 15 mg/kg folic acid, 400 mg/kg choline and 150 μg/kg VB₁₂, from mating until delivery. After farrowing, sows were fed a common lactation diet through a 28‐days lactation period and six litters per treatment were selected to be fed until at approximately 110 kg BW. Maternal MD supplementation resulted in greater birthweight (p < 0.05) and increased the piglet weights (p < 0.01) and litter weights (p < 0.05) at the age of day 28, compared with that in CON group. The offspring pigs in the MD group had greater ADG (p < 0.05) and tended to lower F:G ratio (p = 0.07) compared with that of CON group from day 28 to 180 of age. The offspring pigs from MD group had greater serum IGF‐1 concentrations and expressions of hepatic IGF‐1 gene and muscular IGF‐1 receptor (IGF‐1r) protein at birth (p < 0.05), and greater hepatic IGF‐1 protein (p = 0.03) and muscular IGF‐1r gene expressions (p < 0.05) at slaughter, than that from the CON group. Moreover, the methylation at the promoter of IGF‐1 gene in the liver of newborn piglets and finishing pigs was greater in the MD group than that of the CON group (p < 0.05). In conclusion, maternal MD supplementation throughout gestation could enhance the birthweight and postnatal growth rate of offspring, associated with an increased expression of the IGF‐1 gene and IGF‐1r, as well as the altered DNA methylation of IGF‐1 gene promotor.     

Effect of supplemental β‐carotene compared to retinyl palmitate on fatty acid profile and expression of mRNA from genes involved in vitamin A metabolism in beef feedlot cattle

To examine the effects of dietary β‐carotene (βC) or retinyl palmitate (RP) on fatty acid (FA) profile and mRNA expression, samples were collected from 24 Angus‐cross calves that were allotted to four treatments consisting of RP supplemented at 2200 IU/kg, and synthetic β‐carotene (SβC) supplemented at one, five or 10 times RP. Longissimus muscle (LM) cis‐9, trans‐11 conjugated linoleic acid was greater in RP compared to SβC1X (P = 0.04). The polyunsaturated:saturated FA increased linearly (P = 0.04) in the LM as dietary SβC increased. Expression of βC oxygenase 2 (βCO2), an enzyme that cleaves β‐carotene, was greater in the LM for SβC1X compared to RP and decreased linearly as SβC increased (P ≤ 0.02). Peroxisome proliferator activated receptor γ (PPARγ) expression in the LM increased in SβC1X compared to RP (P = 0.03); however, PPARγ and retinoic acid X receptor α (RXRα) expression decreased linearly (P = 0.02) in the LM with increasing SβC. Retinoic acid receptor α (RARα) expression tended (P = 0.10) to decrease linearly in the LM with increased SβC. In conclusion, SβC supplementation increased mRNA expression of some lipogenic genes in the LM, but increasing dietary SβC inhibited their expression and tended to increase polyunsaturated FA.     

An Investigation of Uterine Nitric Oxide Production in Mares Susceptible and Resistant to Persistent Breeding‐Induced Endometritis and the Effects of Immunomodulation

The first objective of this study was to evaluate intrauterine nitric oxide (NO) and endometrial inducible NO synthase (iNOS) in mares susceptible or resistant to persistent breeding‐induced endometritis (PBIE) within 24 h after breeding. Mares susceptible (n = 6) or resistant (n = 6) to PBIE were inseminated over five cycles, and uterine secretions and endometrial biopsies were collected before and 2, 6, 12 and 24 h after insemination. Uterine secretions were analysed for NO and biopsies were analyzed for iNOS expression. A second experiment evaluated the effect of treatment with dexamethasone or mycobacterial cell wall extract (MCWE) on uterine NO production and endometrial iNOS mRNA expression. Six susceptible mares were inseminated over three cycles with (i) killed spermatozoa without treatment (control), (ii) killed spermatozoa with 50 mg of dexamethasone IV or (iii) MCWE IV 24 h prior to insemination with killed spermatozoa. Six resistant mares were inseminated with killed spermatozoa as a control. Six hours after breeding, uterine biopsies and secretions were collected and evaluated for NO and iNOS mRNA. In Experiment 1, resistant mares had an increase in iNOS mRNA expression 2 h post‐breeding compared to baseline (p = 0.045), 12 h (p = 0.014) and 24 h (p = 0.001). Susceptible mares had higher expression 2 h compared to 6 h (p = 0.046). No differences were observed in mRNA or protein expression of iNOS between resistant and susceptible mares. Resistant mares had a relatively steady amount of total intrauterine NO over 24 h, while susceptible mares had an increase over time, with a significantly higher increase in total NO than resistant mares at 6 (p = 0.04) and 12 h (p = 0.032). In Experiment 2, no differences were observed for iNOS mRNA expression. Susceptible mares had increased NO when compared to resistant mares (p = 0.008) and MCWE decreased NO (p = 0.047).     

Influence of vitamin E on organic matter fermentation,ruminal protein and fatty acid metabolism,protozoa concentrations and transfer of fatty acids

Vitamin E (Vit. E) is discussed to influence ruminal biohydrogenation. The objective of this study was to investigate the influence of a Vit. E supplementation on rumen fermentation characteristics, ruminal microbial protein synthesis as well as ruminal organic matter fermentation. Furthermore, we aimed to investigate the influence of Vit. E supplementation on short‐chain fatty acids (SCFA) and protozoa concentrations in the rumen and, in addition, on transfer rates of middle‐chain and long‐chain fatty acids into the duodenum in lactating dairy cows. Eight rumen and duodenum fistulated German Holstein cows were assigned to either a group receiving 2,327 IU/d Vit. E (138.6 IU/kg DM DL‐α‐tocopherylacetate; n = 4) or a control group (23.1 IU/kg DM; n = 4). Neither ruminal protein synthesis nor organic matter fermentation was influenced by treatment. Vit. E did not act on the concentrations of short‐chain fatty acids and protozoa in rumen fluid. Duodenal flow of C13:0 (1.3 versus 0.2 g/d, p = 0.014) and iso‐C14:0 (1.0 versus 0.5 g/d, p = 0.050) was higher in the Vit. E group. We observed a trend for higher duodenal flows for C12:0 (1.6 versus 0.9 g/d, p = 0.095) and anteiso‐C15:0 (12.2 versus 8.9 g/d, p = 0.084). Transfer rate of C12:0 tended to be higher in the Vit. E group (125.61 versus 73.96, p = 0.082). No other transfer rates were affected by treatment. Further studies are necessary to investigate the influence of Vit. E on rumen microbiota and their fatty acid production as well as on the impact of different doses of Vit. E supplementation on variables of protein synthesis efficiency.     

Effect of dietary Schizochytrium microalga oil and fish oil on plasma cholesterol level in rats

The purpose of the study was to test the hypothesis that the dietary oils with different content of n‐3 polyunsaturated fatty acids (PUFA) eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) affect plasma lipid level in rats in a different degree. The diets with 6% of fish oil (FO) and Schizochytrium microalga oil (SchO; EPA+DHA content in the diets 9.5 + 12.3 and 2.6 + 29.5% of the sum of total fatty acids, respectively) were used; the diet with 6% of safflower oil (high content of n‐6 PUFA linoleic acid, 65.5%; EPA+DHA content 0.7 + 0.9%) was used as a control. The difference between FO and SchO was established only in the case of plasma triacylglycerol (TAG) level: plasma TAG of the FO‐fed rats did not differ from the control rats (p > 0.05), while SchO decreased (p < 0.05) plasma TAG to 46% of the control. On the other hand, FO and SchO decreased (p < 0.05) total plasma cholesterol (TC) in rats in the same extent, to 73% of the control. Regarding the underlying mechanisms for the TC decrease, both SchO and FO up‐regulated hepatic Insig‐1 gene (181 and 133% of the control; p < 0.05), which tended (p = 0.15 and p = 0.19 respectively) to decrease the amount of hepatic nSREBP‐2 protein (61 and 66% of the control). However, neither SchO nor FO influenced hepatic 3‐hydroxy‐3‐methyl‐glutaryl‐CoA reductase gene expression (p > 0.05); SchO (but not FO) increased (p < 0.05) low‐density lipoprotein receptor mRNA in the liver. It was concluded that the decrease of total plasma cholesterol might be caused by an increased cholesterol uptake from plasma into the cells (in the case of SchO), but also by other (in the present study not tested) mechanisms.