Effects of body condition score (BCS) on steroid‐ and eicosanoid‐metabolizing enzyme activity in various mare tissues during winter anoestrus

The objective of this study was to determine the activity of steroid‐ and eicosanoid‐metabolizing enzymes in horses with varying BCSs. The BCSs of twenty non‐pregnant, anoestrous mares were determined prior to euthanasia, and tissue samples were collected from the liver, kidney, adrenal gland, ovary and endometrium. Cytochrome P450 1A (CYP1A), 2C (CYP2C), 3A (CYP3A) and uridine 5′‐diphospho‐glucuronosyltransferase (UGT) activities were determined using luminogenic substrates. The MIXED procedure of SAS was used to test the effect of BCS on enzyme activity and differences between tissues. Activity of CYP1A in adrenals was increased (p ≤ .05) in BCS 5 versus BCSs 4 and 6. Activity of CYP1A in the liver was increased (p = .05) in BCS 4 versus BCSs 5 and 6. Activity of CYP1A was 100‐fold greater (p < .0001) in the liver than in the adrenal, ovary and kidney. Activity of CYP2C was 100‐fold greater (p < .0001) in the liver than in the adrenal, ovary and endometrium. Activity of CYP3A was only detectable in the liver. Activity of UGT in the kidney was decreased (p = .02) in BCS 4 versus BCSs 5 and 6. Activity of UGT was threefold greater (p < .0001) in the liver than in the kidney, whereas activity of UGT was ninefold greater (p < .0001) in the kidney than in the ovary and endometrium. In general, BCS did not alter the activity of steroid‐ and eicosanoid‐metabolizing enzymes in horses. However, tissue differences in these enzymes indicated abundant hepatic metabolism in horses, which is similar to other livestock species.     

Dexamethasone impacts zinc levels in goats by regulating zinc transportation in the colon and the metabolism in the liver

The aim of this study was to investigate the effects of dexamethasone (DEX) on zinc metabolism in goats. In this study, 10 goats were randomly divided into two groups. One group was injected with dexamethasone (Dex group) and the other group was injected with saline (Con group). Dex treatment significantly decreased hepatic zinc levels (p < .01) and increased Zn transporters 1 (ZNT‐1) expression (p < .05). The concentration of zinc in the cecal and colonic contents was significantly increased (p < .05). However, zinc levels were increased only in the colon tissues (p < .05) but not in the cecal tissues (p > .05). A dramatic increase in Zrt‐, Irt‐related proteins 14 (ZIP‐14) expression (p < .05) following Dex treatment was also observed and likely induced the elevated zinc levels in the colon, and a significant reduction in Zip‐14 methylation (p < .05) may be responsible for the observed increase in Zip‐14 expression. Together, these results indicate that Dex influences zinc homeostasis by increasing hepatic ZNT‐1 and colonic ZIP‐14 expression. Additionally, these results provide valuable information for the clinical application of Dex.     

Effects of milk thistle meal on performance,ileal bacterial enumeration,jejunal morphology and blood lipid peroxidation in laying hens fed diets with different levels of metabolizable energy

This study was conducted to evaluate the effects of different levels of milk thistle meal on performance, blood biochemical indices, ileal bacterial counts and intestinal histology in laying hens fed diets containing different levels of metabolizable energy. A total number of 200 Leghorn laying hens (Hy‐Line W‐36) were randomly assigned to eight experimental treatments with five cage replicates of five birds each. Dietary treatments consisted of four levels of milk thistle meal (0%, 15%, 30% and 60%) and two levels of AME_n (11.09 and 12.34 MJ/kg) fed over a period of 80 days. In vitro studies revealed that the total phenolic component of milk thistle meal was 470.64 mg gallic acid equivalent/g of the sample, and its antioxidant activity for inhibiting the 2‐2‐diphenyl‐1‐picrichydrazyl free radical and reducing ferric ions was about 21% higher than that of butylated hydroxyltoluene (p < .05). Diets containing high level of AME_n led to improved egg production (p < .05), egg weight (p < .05), egg mass (p < .01) and feed conversion ratio (p < .01). In addition, offering diets containing high energy significantly enhanced (p < .01) serum triglyceride and malondialdehyde (MDA) concentrations as well as jejunal villus height. Dietary supplementation of 3% milk thistle meal resulted in the best feed conversion ratio (p < .05), reduction of ileal Escherichia coli enumeration (p < .01) and an enhancement in the villus height‐to‐crypt depth ratio (p < .05). Furthermore, feeding incremental levels of this meal led to remarkable decrease in serum cholesterol, triglyceride and MDA (p < .01) concentrations while significant increase in blood high‐density lipoprotein content and goblet cell numbers (p < .05). The present findings indicate that milk thistle meal with high antioxidant and antibacterial properties in laying hen diets may improve health indices and productive performance.     

Exogenous protease supplementation of poultry by‐product meal‐based diets for broilers: Effects on growth,carcass characteristics and nutrient digestibility

An experiment was conducted to investigate the effects of three levels (0%, 3% and 6%) of poultry by‐product meal (PBM) with or without protease on broiler growth, carcass characteristics and nutrient digestibility from 1 to 35 days. Two hundred and forty birds (n = 240) were fed equi‐caloric and equi‐nitrogenous (ME 2850 kcal/kg; CP 20%) diets throughout the experiment. The enzyme supplementation increased feed intake (p < .01) and body weight gain (p < .01), but feed:gain remained unaffected (p > .05) from 1 to 21 days. Increasing level of PBM decreased feed intake (p < .05), but body weight gain was improved (p < .05) at 3% PBM level during 1 to 21 days. The feed:gain was improved (p < .05) in birds fed diets containing 3% PBM. The feed:gain was also improved in birds fed diets containing 3% PBM from 1 to 35 days. However, feed intake and body weight gain in birds fed diets containing PBM remained unaffected. An interaction (p < .01) on feed intake between enzyme and PBM was noticed during 1 to 21 days. However, no interaction was recorded for body weight gain and feed:gain. The per cent carcass yield improved (p < .01) in birds fed diets supplemented with enzyme. The per cent breast meat yield was depressed (p < .005) in birds fed diets containing PBM. Apparent metabolizable energy (p < .001), nitrogen retention (p < .01), apparent metabolizable energy corrected for nitrogen (p < .001), and apparent digestibility coefficient for nitrogen (p < .01) improved in birds fed diets containing enzyme; however, a reverse was noticed in those fed diets containing only PBM. In conclusion, inclusion of 3% PBM along with supplementation of exogenous protease improved performance and nutrient digestibility in broilers.     

Effects and interaction of dietary electrolyte balance and citric acid on growth performance,intestinal histomorphology,digestive enzyme activity and nutrient transporters expression of weaned piglets

Fifty-six piglets were weaned at 21 days and randomly assigned to 1 of 8 dietary treatments with 7 replicate pens for a 14-day experimental period. The eight experimental diets were prepared via a 2 × 4 factorial arrangement with citric acid (CA; 0 and 0.3%) and dietary electrolyte balance (dEB, Na +K − Cl mEq/kg of the diet; −50, 100, 250, and 400 mEq/kg). Varying dEB values were obtained by altering calcium chloride and sodium bicarbonate contents. Dietary CA significantly increased (p < .05) villus height (VH) and villus height:crypt depth (VH:CD) in the jejunum. Piglets fed a 250 mEq/kg diet increased (p < .05) VH and VH:CD values in the duodenum. Jejunal VH and VH:CD increased (quadratic; p < .05), and ileal VH:CD (liner and quadratic; p < .05) decreased as dEB was increased in diets without CA, but no such effect was observed on the diets containing CA (dEB ×CA; p < .05). The CD in jejunum (quadratic; p < .05) increased as dEB was increased in diets containing CA, whereas it was decreased (linear; p < .05) in the diets without CA (dEB ×CA; p < .001). Dietary CA increased maltase activity and reduced the number of Ki67-positive cells (p < .05). Increasing dEB values in diets without CA increased sucrose and lactase activities (quadratic; p < .05), but no such effect was observed in the diets with CA (dEB ×CA; p < .05). An interaction effect between dEB and CA on the number of Ki67-positive cells was observed (p < .001). In conclusion, 250 mEq/kg dEB diet with CA improved piglet intestinal digestion and absorption function by improving intestinal morphology and increasing digestive enzyme activities. However, these improvements were also observed in piglets fed the 100 mEq/kg dEB diet without CA.     

Fertility of Angus cross beef heifers after GnRH treatment on day 23 and timing of insemination in 14‐day CIDR protocol

This study compared artificial insemination pregnancy rate (AI‐PR) between 14‐day CIDR‐GnRH‐PGF2α‐GnRH and CIDR‐PGF2α‐GnRH synchronization protocol with two fixed AI times (56 or 72 hr after PGF2α). On day 0, heifers (n = 1311) from nine locations assigned body condition score (BCS: 1, emaciated; 9, obese), reproductive tract score (RTS: 1, immature, acyclic; 5, mature, cyclic) and temperament score (0, calm; and 1, excited) and fitted with a controlled internal drug release (CIDR, 1.38 g of progesterone) insert for 14 days. Within herd, heifers were randomly assigned either to no‐GnRH group (n = 635) or to GnRH group (n = 676), and heifers in GnRH group received 100 μg of GnRH (gonadorelin hydrochloride, IM) on day 23. All heifers received 25 mg of PGF2α (dinoprost, IM) on day 30 and oestrous detection aids at the same time. Heifers were observed for oestrus thrice daily until AI. Within GnRH groups, heifers were randomly assigned to either AI‐56 or AI‐72 groups. Heifers in AI‐56 group (n = 667) were inseminated at 56 hr (day 32 PM), and heifers in AI‐72 group (n = 644) were inseminated at 72 hr (day 33 AM) after PGF2α administration. All heifers were given 100 μg of GnRH concurrently at the time AI. Controlling for BCS (p < .05), RTS (p < .05), oestrous expression (p < .001), temperament (p < .001) and GnRH treatment by time of insemination (p < .001), the AI‐PR differed between GnRH treatment [GnRH (Yes – 60.9% (412/676) vs. No – 55.1% (350/635); p < .05)] and insemination time [AI‐56 – 54.6% (364/667) vs. AI‐72 – 61.8% (398/644); (p < .01)] groups. The GnRH treatment by AI time interaction influenced AI‐PR (GnRH56 – 61.0% (208/341); GnRH72 – 60.9% (204/335); No‐GnRH56 – 47.9% (156/326); No‐GnRH72 – 62.8% (194/309); p < .001). In conclusion, 14‐day CIDR synchronization protocol for FTAI required inclusion of GnRH on day 23 if inseminations were to be performed at 56 hr after PGF2α in order to achieve greater AI‐PR.     

Predictors of beef calf temperament at weaning and its impact on temperament at breeding and reproductive performance

Two experiments were conducted to determine (i) factors influencing calf temperament at weaning, (ii) association between heifer–calf temperament at weaning and temperament at breeding and (iii) effect of heifer–calf temperament on pregnancy rate per artificial insemination (P/AI). In experiment 1, beef cows and their calves (n = 285) from three farms were used. Sire docility estimated progeny difference (EPD) score, birth type (normal or assisted), calf gender, calf behaviour (during 1st 4 weeks) and calf health status (until weaning) were recorded. Cows and calves were assigned a temperament score (0—calm; 1—excitable), and all cows were given a body condition score (BCS, 1–9; 1—emaciated; 9—obese) at weaning. Calf's illness (p < .05), low sire docility EPD score (p < .05), altered gait (p < .05), altered resting behaviour (p < .01), reduced/no play behaviour (p < .05) and cow excitable temperament (p < .001) increased calf excitable temperament at weaning. In experiment 2, replacement heifer–calves (n = 758) from 12 farms were assigned a temperament score at weaning and later at breeding. Blood from 40 calves at weaning and 31 heifers at initiation of synchronization (same animals) was collected by coccygeal venipuncture for determination of circulating cortisol and substance P concentrations. Heifers were assigned a BCS and reproductive tract score (RTS, 1–5; 1—immature, acyclic; 5—mature, cyclic), synchronized for fixed time AI, observed for oestrus and were artificially inseminated. Cortisol concentrations were increased in excitable heifer–calves compared to calm heifer–calves at weaning (p < .05), and substance P was increased in excitable compared to calm females both at weaning and breeding (p < .05). Low sire EPD docility score (p < .01), heifer–calf excitable temperament at weaning increased excitable temperament at breeding (p < .01). Controlling for BCS categories (p < .01), oestrous expression (p < .0001) and temperament at breeding by oestrous expression (p < .05), the calf's excitable temperament at weaning (p < .001) reduced P/AI (Calm, 62.7 (244/389) vs. Excitable, 53.4% (197/369); p < .01). In conclusion, selection of docile cows and sires with greater docility EPD score should be given consideration to reduce calf excitement. Temperament in beef female can be detected earlier in their life and could be used as a tool in the selection process and to improve their performances.     

Cardiopulmonary and sedative effects of intravenous or epidural methadone in conscious dogs

Cardiopulmonary and sedative effects of intravenous or epidural methadone were compared. Six beagles were randomly assigned to group MIV (methadone 0.5 mg/kg IV + NaCl 0.9% epidurally) or MEP (methadone 0.5 mg/kg epidurally + NaCl 0.9% IV). Cardiopulmonary, blood gas and sedation were assessed at time (T) 0, 15, 30, 60, 120, 240 and 480 min after drug administration. Compared to T0, heart rate decreased at T15–T120 in MIV (p < .001) and T15–T240 in MEP (p < .05); mean arterial pressure was reduced at T15–T60 in MEP (p < .01); respiratory rate was higher at T15 and T30 in both groups (p < .05); pH was lower at T15–T120 in MIV (p < .01) and T15, T30 and T120 in MEP (p < .05); PaCO₂ was higher at T15–T60 in MIV (p < .01) and T15, T30 and T120 in MEP (p < .01); sedation scores were higher at T15 and T30 in MIV and T15–T60 in MEP (p < .05). At T120 and T240, sedation score was higher in group MEP compared with group MIV (p < .01) In conclusion, cardiopulmonary and sedative effects of identical methadone doses are similar when administered IV or epidurally to conscious healthy dogs.     

Effects of polyphenolic extract from Eucommia ulmoides Oliver leaf on growth performance,digestibility, rumen fermentation and antioxidant status of fattening lambs

To investigate the effects of polyphenolic extract from Eucommia ulmoides Oliver leaf (PEEU) on growth performance, digestibility, rumen fermentation and antioxidant status of fattening lambs, 30 weaned male Huzhou lambs were equally divided into three treatments and fed the basal diet supplemented with 0 (CON), 5 (PEEU5) or 10 (PEEU10) g/kg PEEU. Dietary PEEU supplementation did not affect growth performance and apparent digestibility of nutrients. Compared with the CON group, lambs in the PEEU10 group had lower ammonia nitrogen concentration (p < .01) and acetic acid to propionic acid ratio (p < .05) and higher the molar proportion of propionate (p < .05) in rumen fluid. Ammonia nitrogen concentration (p < .01) and the molar proportion of propionate (p < .05) were affected by an interaction between PEEU and sampling day. Dietary PEEU supplementation at 10 g/kg increased total antioxidant capacity (p < .05), superoxide dismutase (p < .05) and glutathione peroxidase (p < .01) activities in serum and glutathione peroxidase activity (p < .05) in liver and decreased (p < .05) malondialdehyde content in serum and liver compared with the CON group. In conclusion, dietary PEEU supplementation affected rumen fermentation patterns and improved antioxidant status of fattening lambs.     

Effects of nanocalcium carbonate on egg production performance and plasma calcium of laying hens

This experiment was conducted to evaluate the effects of nanocalcium carbonate (NCC) instead of calcium carbonate (CC) on egg production, egg weight, egg mass, FCR, blood calcium and egg quality characteristics in laying hens. A total of 120 laying hens were used in a 10‐weeks trial, from week 23 to 33 of age. Laying hens were randomly assigned to six treatments with four replications, five hens each. The experimental treatments involved replacing 50% of the CC in the diet by decreasing amounts of NCC and were T1 Basal diet (BD) with 8.06% CC; T2 (6.045% of CC as a negative control); T3 (4.03% of CC replaced by 2.015% NCC); T4 (4.03% of CC replaced by 1.01% NCC); T5 (4.03% of CC replaced by 0.252% NCC) and T6 (4.03 of CC replaced with 0.126%NCC).Egg weight was unaffected by dietary treatments (p > .05). However, the egg production percentage and egg mass in T6 were less than that of other treatments (p < .05). The laying hens in the control group had the best average feed conversion ratio (p < .05). Also, the lowest concentration of calcium in hens’ blood was recorded for birds fed T6 (p < .05). The best egg shell quality (relative egg shell weight and egg shell weight/surface) was observed in T1 (p < .05).Collectively, our results demonstrated that NCC could replace CC at a lower inclusion level but extreme reduction of calcium concentration in diets (to 1.43% Ca in the T6 group) reduced production performance, egg quality characteristics, Tibia thickness and blood calcium of laying hens.     

Differential gene expression of Eph‐ephrin A1 and LEPR‐LEP with high or low number of embryos in pigs during implantation

The objective of this study was to ascertain whether mRNA and protein expressions of implantation‐related genes (erythropoietin‐producing hepatocellular receptor–ligand A1, Eph‐ephrin A1 and leptin receptor–leptin, LEPR‐LEP) differed between pigs with high and low number of embryos, and whether these differences in gene expression might affect embryo implantation. Experimental pig groups (n = 24) for high and low number of embryos were prepared by altering the number of eggs ovulated in pre‐pubertal gilts treated with 1.5 × (High) or 1.0 × (Low) PG600 ([400 IU PMSG + 200 IU hCG]/dose, AKZO‐NOBEL). Gilts expressing oestrus were artificially inseminated twice and maintained in breeding and gestation until the reproductive tract was collected on day 22 of pregnancy. At slaughter, the reproductive tracts from each pregnant gilt from each treatment were immediately processed to collect samples for RNA and protein analysis. Within each gilt, three conceptus points were sampled, one from each horn and then a random conceptus within the tract. At each conceptus point, endometrial attachment site, chorion–allantois and embryo were collected and immediately frozen in liquid nitrogen. Number of corpus luteum (CL) (35.4 vs. 12.6) and total embryo number (18.8 vs. 10.2) were greater in the high‐embryo compared to the low‐embryo group, respectively (p < .05). Real‐time qPCR results showed that Eph‐ephrin A1 mRNA expression was less in the high‐embryo (p < .05) compared to the low‐embryo group. In addition, Western blotting analysis indicated that Eph‐ephrin A1 and LEP protein expression at endometrial attachment site in high‐embryo was less (p < .05) compared to low‐embryo group. It was also noted that mRNA expression of Eph‐ephrin A1 and LEPR‐LEP was greater in pregnant than non‐pregnant gilts (p < .05). Moreover, mRNA expression of Eph‐ephrin A1 (p < .05) and LEPR‐LEP was greatest at endometrial attachment site among all three tissues. There was a positive correlation between expressions of Eph‐ephrin A1, LEPR‐LEP and embryo length with the correlation coefficient 0.31–0.59. For Eph‐ephrin A1, the highest correlation coefficient appeared between Eph A1 expression and normal embryo number, between ephrin A1 expression and embryo length. For LEPR‐LEP, the highest correlation coefficient appeared between LEPR‐LEP expression and ovary weight (0.79 for both, p < .05), followed by embryo length and weight. The results of this study suggest that low expression of Eph‐ephrin A1 and LEPR‐LEP is somehow related to increased embryo number during implantation and that endometrial attachment site might be the main target tissue of these gene products. Yet, the increased expression of Eph‐ephrin A1 and LEPR‐LEP appeared associated with increased embryo growth (length and weight) and ovary weight, Eph‐ephrin A1 and LEPR‐LEP might play roles in the regulation of embryo implantation in pigs.     

Effect of lactic acid bacteria‐treated King grass silage on the performance traits and serum metabolites in New Zealand white rabbits (Oryctolagus cuniculus)

This study examined the effects of Lactobacillus plantarum and Pediococcus acidilactici‐treated silage of King grass (Pennisetum purpureophoides) on the productive traits and blood biochemistry of New Zealand white (NZW) rabbits. King grass was ensiled without or with L. plantarum (T1), P. acidilactici (T2) and P. acidilactici + L. plantarum (T3). A total of 72 male NZW rabbits (6 weeks) of similar weight and appearance were housed in groups in metallic cages in a completely randomized design. The performance traits, daily feed intake, body weight gain and feed convention ratio were not affected by the silage treatment. Triglyceride and alanine aminotransferase (ALT) concentration was increased (p < .05) in T2 (0.92 mmol/L) compared to the other treatments. The superoxide dismutase (SOD), phosphorous (P) and magnesium (Mg) increased (p < .05) in T1 (658.01 nmol/ml, 5.1 mg/kg and 2.43 mg/kg, respectively) compared to the control and other experimental groups. Cu and Mn decreased (p < .05), while Zn increased (p < .05) in the treated groups compared to the control. The CP decreased (p < .05) in the treatment groups compared to the control. Neutral detergent fibre (NDF) increased (p < .05) in T2 (54.40%) compared to the other treatments. The inoculation of lactic acid bacteria‐treated King grass silage supplementation positively affected rabbit performance and improved blood cholesterol profile, antioxidant status and improve nutrients digestibility.     

Evaluation of haemodynamic changes of uterine arteries using Doppler ultrasonography during different stages of pregnancy in Bos indicus cows

The objectives of the study were to evaluate haemodynamic changes and their relationships among ipsilateral (IPS) and contralateral (CONT) uterine arteries (UA) during different stages of pregnancy in Bos indicus cows. Multiparous pregnant cows (n = 40) having a gestation length 30.47 ± 0.54 (mean ± SD) days were randomly enrolled and subjected to Doppler ultrasonography sequentially at 1st, 2nd, 4th, 6th and 8th months of gestation. Blood flow indices including diameter of UA (mm), blood flow volume (BFVo, ml/min), blood flow velocity (BFVe, cm/s), time-averaged maximum velocity (TAMV, cm/s), pulsatility index (PI) and resistance index (RI) were recorded. Data were analysed with mixed models using the PROC MIXED procedures, and Pearson correlation coefficients were calculated using the PROC CORR statement in SAS. The final statistical models included the fixed effects of side of UA, gestation month and the interaction between side of UA and gestation month. Results revealed that the mean diameter of the UA (12.13 ± 0.22 vs. 10.09 ± 0.22), BFVo (1236.33 ± 0.55 vs. 770.41 ± 0.55), BFVe (17.18 ± 0.42 vs. 15.58 ± 0.42) and TAMV (17.11 ± 0.44 vs. 15.77 ± 0.44) was higher (p < .05) in IPS as compared to CONT side of the UA in cows. However, PI and RI did not differ between IPS and CON arteries of uterus in cows. A very high and positive correlation (r = .89; p < .05) existed between the diameter of UA and BFVo starting from 1st to 8th months of gestation in IPS as well as CONT sides of UA. Moreover, TAMV was highly and positively correlated (r = .91; p < .05) with BFVe throughout the gestation. In conclusion, these haemodynamic changes in the UA could be used as a valuable validity tool to differentiate the compromised pregnancy in Bos indicus cows.     

Carcass traits,meat quality,antioxidant status and antioxidant gene expression in muscle and liver of Hu lambs fed perilla seed

The effects of perilla (Perilla frutescens L.) seed on carcass traits, meat quality, antioxidant status and antioxidant gene expression in the liver and muscle of Hu lambs were investigated in this study. Sixty Hu lambs (23.02 ± 1.36 kg) were randomly divided into four experimental groups receiving diets containing 0%, 5%, 10% or 15% perilla seed (CD, 5%PFSD, 10%PFSD and 15%PFSD, respectively). The addition of perilla seed had no significant impacts on carcass traits (p > .05). There were no differences in pH, meat colour, drip loss, cooking loss or shear force among the four treatments (p > .05). Addition of perilla seed increased (p < .05) deposition of intramuscular lipids but had no effect on other chemical components in the longissimus dorsi (LD) (p > .05). The 15%PFSD diet decreased the total antioxidant capacity (T‐AOC) and malondialdehyde (MDA) content in the liver (p < .05 for both) but increased the activity of these antioxidant enzymes in LD (p < .05 for both). Compared to CD, addition of perilla seed increased superoxide dismutase (SOD) and glutathione peroxidase (GPX) expression in the liver and LD (p < .05 for all). These results indicate that perilla seed supplementation in lambs’ diets can increase deposition of intramuscular lipids and improve muscular oxidative status and meat quality.     

m‐carboxycinnamic acid bishydroxamide improves developmental competence,reduces apoptosis and alters epigenetic status and gene expression pattern in cloned buffalo (Bubalus bubalis) embryos

Incomplete or aberrant reprogramming of nuclear genome is one of the major problems in somatic cell nuclear transfer. In this study, we studied the effect of histone deacetylase inhibitor m‐carboxycinnamic acid bishydroxamide (CBHA) on in vitro development of buffalo embryos produced by Hand‐made cloning. Cloned embryos were treated with CBHA (0, 5, 10, 20 or 50 μM) for 10 hr from the start of reconstruction till activation. At 10 μM, but not at other concentrations examined, CBHA increased (p < .05) the blastocyst rate (63.77 ± 3.97% vs 48.63 ± 3.55%) and reduced (p < .05) the apoptotic index of the cloned blastocysts (8.91 ± 1.94 vs 4.36 ± 1.08) compared to untreated controls, to levels similar to those in IVF blastocysts (4.78 ± 0.74). CBHA treatment, at all the concentrations examined, increased (p < .05) the global level of H3K9ac in cloned blastocysts than in untreated controls to that observed in IVF blastocysts. Treatment with CBHA (10 μM) decreased (p < .05) the global level of H3K27me3 in cloned blastocysts than in untreated controls but it was still higher (p < .05) than in IVF blastocysts. CBHA (10 μM) treatment increased (p < .05) the relative expression level of pluripotency‐related genes OCT‐4 and NANOG, and anti‐apoptotic gene BCL‐XL, and decreased (p < .05) that of pro‐apoptotic gene BAX than in untreated controls but did not affect the relative expression level of apoptosis‐related genes p53 and CASPASE3 and epigenetics‐related genes DNMT1, DNMT3a and HDAC1. These results suggest that treatment of cloned embryos with 10 μM CBHA improves the blastocyst rate, reduces the level of apoptosis and alters the epigenetic status and gene expression pattern.     

The effect of maternal obesity on fatty acid transporter expression and lipid metabolism in the full‐term placenta of lean breed swine

This study was conducted to evaluate the influence of back‐fat thickness (BF), at mating of sows, on the maternal and newborn circulating lipids, expression of placental fatty acids (FA) transporters and lipid accumulation in placenta. Full‐term placentas were obtained by vaginal delivery from BFI (9–14 mm; n = 37), BFII (15–19 mm; n = 43) and BFIII (20–27 mm; n = 38) sows according to BF at mating, and frozen placental sections were analysed for fat accumulation. Blood samples were collected from the sows of day 105 pregnancy and from cord blood at delivery. mRNA and protein expression levels were evaluated with real‐time RT‐PCR and Western blotting. Our results demonstrated that BFII females had significantly increased litter weight and placental efficiency, decreased maternal triglyceride (TG) and non‐esterified fatty acids (NEFA) levels, decreased maternal IL‐6, TNFα and leptin levels compared to BFIII females (p < .05). BFIII sows were associated with significantly decreased newborn TG levels, increased newborn glucose, IL‐6 and TNFα levels compared to BFI or BFII sows (p < .05). BFI and BFII females had significantly decreased placental TG, NEFA and cholesterol (CHOL) contents compared to BFIII females (p < .05). Moreover, decreased CD36, FATP1, FABP4, and FABP1 mRNA and protein and FATP4 protein expression, and increased LPL activity were also observed in BFIII group compared with BFII group (p < .05). PPARγ mRNA and protein and lipogenic genes such as SREBP‐1c, ACSL1, ACCα, FAS and SCD mRNA expression were downregulated or upregulated, respectively, in the placentas of BFIII sows compared to BFI or BFII sows (p < .05). Overall, this study demonstrated that there is no advantage, in terms of litter live size, litter weight and placental FA transport and metabolism, in performing the mating of sows with BF>19 mm.     

Effect of Oestrus Synchronization with PGF2α/eCG/hCG on Luteal P4 Synthesis in Early Pregnant Gilts

Administration of hormones to synchronize oestrus is a useful tool in animal breeding. However, exogenous ovarian stimulation may be detrimental to reproductive function. This study was aimed to examine whether an oestrus synchronization with PGF2α/eCG/hCG could affect luteal P4 synthesis in early pregnant gilts. Corpora lutea (CLs) were collected on days 9, 12 and 16 of pregnancy from gilts with natural (n = 16) and synchronized (n = 18) oestrus and analysed for (i) the expre‐ssion of steroidogenic acute regulatory protein (StAR), cytochrome P450 family 11 subfamily A polypeptide (CYP11A1), and 3β‐hydroxysteroid dehydrogenase (3βHSD); (ii) the concentration of P4 in the luteal tissue and blood; and (iii) the expression of luteinizing hormone receptors (LHR) and oestrogen receptors (ERα and ERβ). Additionally, the effect of LH on P4 secretion from CL slices collected from synchronized and naturally ovulated animals has been studied in vitro. PGF_2α/eCG/hCG administration increased mRNA expression of StAR, CYP11A1, 3βHSD, and LHR on day 9 and CYP11A1 and LHR on day 12 of pregnancy compared with the control group (p < 0.05). CYP11A1, 3βHSD, LHR, ERα and ERβ proteins were not affected by synchronization; only StAR protein increased in hormonally treated animals (p = 0.017). The concentration of P4 in luteal tissue was greater on day 9 (p < 0.01), but lower on day 16 (p < 0.05) in gilts with hormonally induced oestrus compared with control animals. Blood serum levels of P4 were lower in synchronized than control gilts (p < 0.001). Synchronization did not affect LH‐stimulated P4 secretion from luteal slices; however, greater basal concentration of P4 in incubation medium was detected for CLs collected from synchronized than control gilts (p < 0.05). In conclusion, synchronization of oestrus with PGF2α/eCG/hCG protocol in gilts did not impair the expression of luteal P4 synthesis system, although decreased P4 concentration in the blood.