Maternal obesity stimulates lipotoxicity and up‐regulates inflammatory signaling pathways in the full‐term swine placenta

This study aimed to investigate the effects of back‐fat thickness (BF), at mating of sows, on placental lipotoxicity, oxidative stress, and inflammation. We performed iTRAQ labeling‐based proteomic analysis on term placentas obtained by vaginal delivery from BFI (15–20 mm, control) and BFII (21–27 mm, obese) sows formed according to BF at mating. Proteomic analysis revealed 413 proteins to be significantly different in placenta from BFII sows by ≥1.2‐fold. Gene ontology (GO) analysis identified proteins related to lipid metabolism and inflammatory response to be altered in placenta from obese sows. Indicative of a lipotoxic placental environment, increased placental lipid, and up‐regulated mRNA expression of lipogenic genes, including ADRP (p = .06), PPARD, FASN, ACACA, DGAT1, and LIPIN3, were associated with decreased AMPK and increased activation of WNT signaling in placenta from BFII group (p < .05). Furthermore, we observed a 18% decrease in total antioxidant capacity (TAC), increased mRNA content of pro‐inflammatory cytokines IL‐6, IL‐18, and TNF‐α, and increased activation of inflammatory NF‐κB and JNK signaling in placenta from BFII sows that was significantly associated with macrophage accumulation (p < .05). These findings suggest that maternal obesity aggravates a lipotoxic environment in pig term placenta that may be associated with placental dysfunction and impaired fetal growth.     

Maternal dietary linoleic acid supplementation promotes muscle fibre type transformation in suckling piglets

As meat quality is basically dependent on muscle fibre characteristics, it is important to know how muscle fibres are regulated and transformed. This study aimed to investigate the effect of maternal dietary supplementation on muscle fibre types using 3% saturated fatty acid (palmitic acid, PA) or 3% unsaturated fatty acid (linoleic acid, LA) from 80 days of gestation to the weaning of offspring (25 days post‐natal). The results indicated that higher mRNA levels of MyHCI type genes were found in the soleus muscles of piglets that suckled from LA‐supplemented sows than from PA‐supplemented sows. In addition, LA treatment increased the gene expression of the type I muscle fibre marker troponin I (p < 0.01), suggesting that LA promoted muscle fibre type transformation to type I fibres. Moreover, PGC‐1α (p < 0.01) and MEF2c (p < 0.05) mRNA levels were higher in the piglets from the LA treatment group than in those from the PA treatment group. Furthermore, LA supplementation also significantly increased AMP‐activated protein kinase (AMPK) mRNA levels (p < 0.05), which is an upstream regulator of PGC‐1α. Collectively, these findings demonstrated that maternal dietary LA supplementation promoted muscle fibre transformation to type I fibre and that this process may be mediated through an AMPK‐dependent pathway.     

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.     

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.     

Evaluation of three different patterns of feed intake during early lactation in lactating sows

Forty‐five multiparous sows (Landrace × Yorkshire; parity: 3.58 ± 1.30) were used to determine the effects of three patterns of feed intake during early lactation on the performance of lactating sows. Experimental treatments were as follows: IFI‐1.4, the amount of feed increased by 1.4 kg per day for the first 5 days post‐farrowing, followed by ad libitum feeding until weaning; IFI‐1.0, the amount of feed increased by 1.0 kg per day for the first 7 days post‐farrowing, followed by ad libitum feeding until weaning; IFI‐0.7, the amount of feed increased by 0.7 kg per day for the first 10 days post‐farrowing, followed by ad libitum feeding until weaning. The number of live piglets at birth/litter in three dietary treatments was 11.50, 10.07, and 10.85, respectively. Sows in the IFI‐1.4 treatment had lower backfat loss during lactation, greater daily feed intake during days 1–6, 7–12, and 1–24 compared with those in the IFI‐0.7 treatment (p < .05). The litter weaning weight, litter weight gain, and average litter daily gain in the IFI‐1.4 treatment were greater compared with those in the IFI‐0.7 treatment (p < .05). In conclusion, the results indicated that the IFI‐1.4 feed intake pattern allowed lactating sows and their litters to obtain optimal performance.     

An investigation into uterine capacity based on litter and placental characteristics in two sow lines with different prolificacy (Danish Landrace x Danish Yorkshire versus German Saddleback)

Litter size in modern so called hyperprolific pig (Sus scrofa Linnaeus) breeds such as of crossbred Danish Landrace x Danish Yorkshire (LY/YL) sows increased remarkably over recent years, however, commonly associated with reduced piglet birth weight and higher within litter birth weight variability likely due to a limited uterine capacity. Since investigation into this issue is patchy, the aim of this study was to investigate uterine capacity based on litter and placental characteristics in two sow lines with different prolificacy, that is crossbred Danish genetic (Danish Landrace x Danish Yorkshire; DG; n = 14) and purebred German Saddleback (GS) sows (n = 12). Parameters recorded were litter size, piglet birth weight and vitality, placental weight and surface area as well as placental vascularization. Litters of DG were on average larger than of GS (p < .001). Piglets of DG weighed on average less than GS (p < .001) and were less vital (p < .001–.142). Increasing litter size was associated with reduced piglet birth weight and increased within litter birth weight variability in GS, but not in DG. DG had on average a lower placental weight (p < .001) and smaller placentae (p < .001) than GS, but the placenta was on average more efficient than of GS (based on the quotient of piglet and corresponding placental weight; p < .001). Vascularization of placentae was on average not or only slightly different between breeds (p < .05 – .982). Remarkably, however, vascularization of the lateral and apical chorionic epithelium of the chorionic ridges as the immediate foetal/maternal interface was on average slightly higher in DG than GS (p < .05–.111). Results thus demonstrate that uterine capacity based on litter and placental characteristics is higher in DG than GS sows.     

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 Inulin Supplementation in Low‐ or High‐Fat Diets on Reproductive Performance of Sows and Antioxidant Defence Capacity in Sows and Offspring

This experiment was conducted to investigate the effects of inulin supplementation in low‐ or high‐fat diets on both the reproductive performance of sow and the antioxidant defence capacity in sows and offspring. Sixty Landrace × Yorkshire sows were randomly allocated to four treatments with low‐fat diet (L), low‐fat diet containing 1.5% inulin (LI), high‐fat diet (H) and high‐fat diet containing 1.5% inulin (HI). Inulin‐rich diets lowered the within‐litter birth weight coefficient of variation (CV, p = 0.05) of piglets, increased the proportion of piglets weighing 1.0–1.5 kg at farrowing (p < 0.01), reduced the loss of body weight (BW) and backfat thickness (BF) during lactation (p < 0.05) and decreased the duration of farrowing as well as improved sow constipation (p < 0.05). Sows fed fat‐rich diets gained more BW during gestation (p < 0.01), farrowed a greater number of total (+1.65 pigs, p < 0.05) and alive (+1.52 pigs p < 0.05) piglets and had a heavier (+2.06 kg, p < 0.05) litter weight at birth as well as a decreased weaning‐to‐oestrous interval (WEI, p < 0.01) compared with sows fed low‐fat diets. However, it is worth noting that the H diet significantly decreased the serum activities of superoxide dismutase (T‐SOD) and glutathione peroxidase (GSH‐Px) and increased the serum malondialdehyde (MDA) levels in sows and piglets (p < 0.05). In contrast, HI diet enhanced the activities of T‐SOD and GSH‐Px and decreased the serum MDA concentrations (p < 0.05) in sows and piglets. In summary, the fat‐rich diets fed to sows during gestation had beneficial effects on reproductive performance, but aggravated the oxidative stress in sow and piglets. Inulin‐rich diets fed to sow during gestation had beneficial effects on within‐litter uniformity of piglet birthweight and enhanced the antioxidant defence capacity of sows and piglets.     

Effects of Saccharomyces cerevisiae supplementation on growth performance,plasma metabolites and hormones,and rumen fermentation in Holstein calves during pre‐ and post‐weaning periods

This study aimed to evaluate the effects of supplementing Saccharomyces cerevisiae (SC) during the pre‐ and post‐weaning periods on growth, metabolic and hormonal responses, and rumen fermentation in calves. Three‐week‐old Holstein calves were assigned to either control (n = 12) or SC group (n = 12), the latter of which received 2 × 10⁹ cfu/day of SC. The experiment was conducted over a period of 7 weeks around weaning. Daily gain (DG) in the SC group was higher (p < .05) than that in the control group. In the SC group, plasma glucose, insulin, and growth hormone (GH) concentrations were higher (p < .05) and concentrations of glucagon and insulin‐like growth factor 1 (IGF‐1) tended to be higher (p < .1) than in the control group. Proportion of rumen propionate and concentration of rumen ammonia nitrogen at 10 weeks of age were greater (p < .05) in the SC group than that in the control group. Supplementation of SC around weaning may improve dietary nutrient and energy availability and increase plasma GH and IGF‐1 concentrations. These changes observed in SC‐supplemented calves could be closely related to the improvement of DG.     

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.     

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.     

Effects of dietary resveratrol supplementation during gestation and lactation of sows on milk composition of sows and fat metabolism of sucking piglets

The purpose of this article was to investigate the effects of dietary resveratrol supplementation during gestation and lactation of sows on the milk composition of sows and the fat metabolism of sucking piglets. Forty sows were allotted to two experimental treatment groups that included the following: (a) control sows (CON treatment, n = 20) fed with a corn–soybean meal control diet and (b) treatment sows (RES treatment, n = 20) fed with a control diet with addition of 300 mg/kg resveratrol. The results showed that the content of lactose in the colostrum was increased (p < 0.05) and the content of fat in 21‐day milk was increased (p < 0.05) by dietary resveratrol supplementation. In the RES treatment group, the concentrations of high‐density lipoprotein‐cholesterol (HDL‐C), low‐density lipoprotein‐cholesterol (LDL‐C), lipase activity and insulin (INS) in plasma of sucking piglets were increased (p < 0.05). In the adipose tissue, the enzyme activities of hormone‐sensitive lipase (HSL), acetyl‐CoA carboxylase (ACC) and lipoprotein lipase (LPL) increased significantly by RES treatment (p < 0.05), and the mRNA levels of acetyl coenzyme A‐alpha (ACCα), LPL, fatty acid transport protein (FATP1) and CCAAT–enhancer‐binding protein gene (C/EBPα) were higher in the RES treatment group (p < 0.05). In conclusion, resveratrol supplementation on gestational and lactating sows improved the content of lactose in the colostrum and the content of fat in milk at day 21 of lactation. In addition, resveratrol supplementation on sows increased HDL and LDL in the plasma of piglets. In piglet adipose tissue, the enzyme activity and mRNA level related to lipolysis, fatty acid uptake from circulating triacylglycerols and lipogenesis are partially improved by resveratrol supplementation on sows. These aspects affect fat metabolism in piglets.     

Effects of deficiency and surplus dietary threonine on reproductive performance of primiparous pregnant gilts

Dietary threonine imbalance is known to impair reproductive performances of gestating sows, but the underlying mechanisms are largely unknown. In this study, effects of deficiency and surplus dietary threonine during gestation on reproductive performance, serum metabolites and hormones concentration, and colostral nutrient and immunoglobulin contents of primiparous sows were investigated. Ninety primiparous pregnant gilts were assigned to one of the three dietary treatments with different standardized ileal digestible threonine/lysine ratios at 0.59, 0.72 and 0.85, which represented deficient (DT), adequate (AT) and surplus (ST) dietary threonine concentration respectively. Maternal body weight gain from day 80–110 of gestation was highest (p < .05) for gilts fed AT than for gilts fed DT or ST. On days 30 and 110, serum threonine concentration increases in a dose‐dependent manner with the increasing of dietary threonine concentration in (p < .01), serum urea nitrogen concentration was lower (p < .01) in gilts fed AT than DT or ST, and serum insulin‐like growth factor‐I (IGF‐I) was lowest (p < .05) for gilts fed DT. On day 110, gilts fed AT had lower serum progesterone concentration but higher concentrations of serum prolactin (p < .05) compared to DT and ST. Concentration of colostral immunoglobulin A and G from gilts fed DT was lower (p < .05) compared with gilts fed AT or ST. In conclusion, gilts with the adequate threonine intake were more able to conserve dietary amino acids to support foetal and maternal tissue gain. Deficient or ST threonine intake may induce a delay in changes in progesterone and prolactin concentrations during the prepartum period impeding the transition from pregnancy to lactation.     

Effectiveness of controlled internal drug release device treatment to alleviate reproductive seasonality in anestrus lactating or dry Barki and Rahmani ewes during non‐breeding season

This study aimed to evaluate the effectiveness of hormonal treatments on ovarian activity and reproductive performance in Barki and Rahmani ewes during non‐breeding season. Forty‐eight multiparous ewes, 24 Barki and 24 Rahmani ewes were divided into two groups, 12 lactating and 12 dry ewes for each breed. Controlled internal drug release (CIDR) device was inserted in all ewes for 14 days in conjunction with intramuscular 500 IU equine chronic gonadotrophin (eCG) at day of CIDR removal. Data were analysed using PROC MIXED of SAS for repeated measures. Breed, physiological status and days were used as fixed effects and individual ewes as random effects. Barki ewes recorded higher (p < .05) total number of follicles, number of large follicles, serum estradiol concentration and estradiol: progesterone (E₂:P₄) ratio compared to Rahmani ewes. Lactating ewes recorded higher (p < .05) number of small follicles and lower concentration of total antioxidant capacity (TAC) compared to dry ewes. Number and diameter of large follicles recorded the highest (p < .05) values accompanied with disappearance of corpora lutea at day of mating. Serum progesterone concentration recorded lower (p < .05) value at day of mating and the highest (p < .05) value at day 35 after mating. CIDR‐eCG protocol induced 100% oestrous behaviour in both breeds, but Rahmani ewes recorded longer (p < .05) oestrous duration compared to Barki. Conception failure was higher (p < .05) in Barki compared to Rahmani ewes. In conclusion, CIDR‐eCG protocol was more potent in improving ovarian activity in Barki compared to Rahmani ewes, but this protocol seems to induce hormonal imbalance in Barki ewes that resulted in increasing conception failure compared to Rahmani ewes.     

Effects of tail docking and/or teeth clipping on behavior,lesions, and physiological indicators of sows and their piglets

To evaluate effects of tail docking and/or teeth clipping on sows and their piglets, a total of 24 sows and their 302 piglets at 3 days of age were randomly allocated to one of four treatments: teeth clipping and tail docking (TCTD), teeth clipping (TC), tail docking (TD), or intact teeth and tail (Intact). Behavior of piglets and sows, lesions on the body and tail of piglets and sows' teats were inspected. Heart rates of processed piglets were increased (p < .01) during the procedures. Teeth clipping decreased body surface temperature (p < .01) of piglets during and after the procedures but tail docking did not (p > .01). Processed piglets spent more (p < .05) time lying alone and playing/fighting than sham‐processed piglets. Tail docked piglets spent less (p < .01) time standing than tail sham‐docked piglets. Intact teeth increased (p < .05) the avoidance behaviors of sows. Teeth clipping decreased (p < .05) the lesion scores on the anterior, middle, and posterior teats. Taken together, piglet teeth clipping had more impact on sows and their piglets than tail docking did in the lactation period based on our findings.     

Effects of voltage strength during electroporation on the development and quality of in vitro‐produced porcine embryos

This study was conducted to determine suitable conditions for an experimental method in which the CRISPR/Cas9 system is introduced into in vitro‐produced porcine zygotes by electroporation. In the first experiment, when putative zygotes derived from in vitro fertilization (IVF) were electroporated by either unipolar or bipolar pulses, keeping the voltage, pulse duration and pulse number fixed at 30 V/mm, 1 msec and five repeats, respectively, the rate of blastocyst formation from zygotes electroporated by bipolar pulses decreased compared to zygotes electroporated by unipolar pulses. In the second experiment, the putative zygotes were electroporated by electroporation voltages ranging from 20 V/mm–40 V/mm with five 1‐msec unipolar pulses. The rate of cleavage and blastocyst formation of zygotes electroporated at 40 V/mm was significantly lower (p < .05) than that of zygotes electroporated at less than 30 V/mm. Moreover, the apoptotic nuclei indices of blastocysts derived from zygotes electroporated by voltages greater than 30 V/mm significantly increased compared with those from zygotes electroporated by voltages less than 25 V/mm (p < .05). When zygotes were electroporated with Cas9 mRNA and single‐guide RNA (sgRNA) targeting site in the FGF10 exon 3, the proportions of blastocysts with targeted genomic sequences were 7.7% (2/26) and 3.6% (1/28) in the embryos derived from zygotes electroporated at 25 V/mm and 30 V/mm, respectively. Our results indicate that electroporation at 25 V/mm may be an acceptable condition for introducing Cas9 mRNA and sgRNA into pig IVF zygotes under which the viability of the embryos is not significantly affected.     

Effects of 25‐hydroxycholecalciferol supplementation in maternal diets on reproductive performance and the expression of genes that regulate lactation in sows

One hundred Yorkshire × Landrace sows were randomly assigned to one of two dietary treatments (diet ND: 6,000 IU vitamin D₃/d feed; diet 25‐D: 200 μg/day 25OHD₃ feed). The experiment began on d 90 of gestation and continued until weaning on day 21 of lactation. In sows that received 25OHD₃, the growth rate of the piglets before weaning was significantly accelerated (0.266 kg/day, p < .05). Sow serum was collected after weaning, and those in the 25OHD₃ group were found to have significantly higher serum calcium (CA) and phosphorus (PI) levels (p < .05). Interestingly, the oestrus cycle of sows fed 25OHD₃ was significantly shortened (p < .05), the oestrus time was concentrated on the fifth day after weaning, and the piglets were born with a higher degree of uniformity (p < .05). Colostrum was collected on the day of delivery, and the colostrum of sows fed 25OHD₃ contained higher milk fat content than the control group (p < .05). 25OHD₃ supplementation increased the mRNA and protein expression of INSIG1 and SREBP1, which regulate milk fat synthesis, in the mammary gland of lactating sows (p < .05). In conclusion, 25OHD₃ supplementation in maternal diets improved reproductive performance, milk fat content and the mRNA and protein levels of genes regulating milk fat synthesis in lactating sows.     

Ovine sperm DNA oxidation quantification using an 8‐OHdG immunodetection assay

Our aim was to optimize 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG) immunodetection in order to detect DNA damage caused by oxidative stress that may not be detected by other DNA integrity analysis techniques, especially due to the high compaction of DNA in ruminants. Semen samples from 6 rams were cryopreserved. After thawing, samples were subjected to the DNA oxidation quantification using an 8‐OHdG immunodetection assay by flow cytometry. We have evaluated two different incubation times (30 min vs. overnight) at 4°C of the primary antibody (monoclonal anti‐8‐OHdG antibody). We have also compared the results of this technique with the sperm chromatin structure assay (SCSA^®). The analysis revealed that there were no significant differences (p > .05) between different incubation times. However, overnight incubation seems to cause more non‐specific binding of the secondary antibody. Significant differences (p < .05) between subjects and oxidation controls (8 M H₂O₂/800 μM FeSO₄?7H₂O) were evident. We can conclude that the 8‐OHdG immunodetection assay for DNA oxidation quantification of ram sperm can be performed subjecting sperm samples to a very high oxidative treatment.     

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.