Pre- and post-weaning nutrition status affects sheep mammary gland morphology and promotes differential gene expression.

Little is known about how varying the plane of nutrition before and after weaning can influence gene expression that drives mammary gland development (MGD). Therefore, the aim of this study was to investigate this paradigm in a seasonal sheep breed. Forty pre-weaning 30-day old Kurdish female lambs were fed either a low-nutrient and protein (L; 2.0 Mcal ME/kg DM; 8.70% crude protein [CP]) or high-nutrient and protein diet (H; 2.5 Mcal ME/kg DM; 14.80% CP). At d 120 (weaning), lambs were allocated into a 2 × 2 arrangement based on pre- and post-weaning diets, resulting in four groups (LL, LH, HL, HH). On d 210, mammary biopsy samples were taken for histomorphological and gene expression studies. Alveoli were larger in HH compared with all other groups (p < 0.05). Whilst estrogen receptor-alpha, progesterone receptor, growth hormone receptor and insulin-like growth factor binding protein-1 gene expression was modulated depending on pre- or post-weaning nutritional levels (p < 0.01). Overall, a consistent high plane of nutrition promoted MGD, with more a complex relationship evident between the expression of genes critical for mammary gland function and development. These findings provide scope for future tailored nutritional strategies to optimize both liveweight gain and MGD.

Monensin supplementation downregulated the expression signature of genes involved in cholesterol synthesis in the ruminal epithelium and adipose tissue of lambs.

To understand the metabolic mechanisms regulating lipid metabolism by monensin, Afshari male lambs (n = 16) with 41.0 ± 2.4 kg body weight (BW, mean ± SD) at approximately 180 days of age were randomly assigned equally to two dietary treatments. After a 21-day pre-adaptation period, all animals in two groups continued to receive the basal diet, but one group received no monensin supplementation (control) while the other group received 30 mg/day of monensin per animal. Individual BW was recorded weekly to determine the average daily body weight gain (ADG). At the end of the 56-day experimental period, lambs were weighed and slaughtered. Monensin supplementation did not affect BW, ADG, and rumen fermentation characteristics. However, monensin significantly downregulated the sterol regulatory element-binding protein (SREBP)-2 gene expression in all sample tissues (p < 0.05). Also, monensin downregulated expressions of SREBP-1c and peroxisome proliferator-activated receptor (PPAR)-γ in back fat tissues. Monensin increased the expression of 3-hydroxy-3-methylglutaryl-CoA synthase (HMGCS)-2, but it decreased the mRNA abundance of HMGCS-1 in the rumen epithelial tissues (p < 0.05). Our data suggest that monensin downregulates cholesterol synthesis via inhibition of HMGCS-1 and impairment of the SREBP pathway, probably due to a crosstalk among different tissues to control energy metabolism.

Short communication: effect of dietary supplementation with a mixture of fish and sunflower oils on the expression of key lipogenic and cholesterologenic genes in adipose tissues with different metabolic functions.

The present study investigated the effects of dietary fish and sunflower oils as sources of n-3, n-6 polyunsaturated fatty acids (PUFA) on the expression of key lipogenic and cholesterologenic genes in subcutaneous adipose tissue (SAT) and tail adipose tissue (TAT) of fat-tailed sheep. Twenty-six male Afshari lambs were divided into 4 groups. Three groups were fed a high concentrate basal diet plus 100 g/lamb/day oil supplement (OS; 60 g sunflower oil and 40 g fish oil) beyond a 21-day adaptation period for 10, 20, and 30 days (groups OS10, OS20, and OS30; n = 6, each) until slaughter. A control group was slaughtered at the last day of adaptation (OS0; n = 4). Expression of PPARγ, SREBP-1c, and SREBP-2 were determined in TAT and SAT. All transcription factors had lower expression in SAT than TAT. Feeding OS induced a similar pattern of SREBP-1c expression in both TAT and SAT with highest values in OS20. SREBP-2 mRNA decreased by > 50% in TAT of OS30 compared to OS0, whereas the expression of SREBP-2 mRNA did not change in SAT in the same period. PPARγ expression was not affected over time either in SAT or TAT. Plasma concentrations of cholesterol and blood urea nitrogen increased in OS20. The comparison of gene expression responses to OS in TAT vs. SAT suggest that PUFA-mediated effects on lipid metabolism differ between SAT and TAT, which may be linked to the specific role of TAT in energy and water balance under arid conditions.

Moderate overfeeding of different sources of metabolizable energy and protein. II: Effects on inflammatory status of sheep in late pregnancy and growth trajectory of the offspring.

Our objective was to evaluate the effects of moderate overfeeding of different sources of metabolizable energy (ME) and protein (MP) on inflammatory status of sheep in late gestation and short-term growth performance of the offspring. A total of 32 healthy pregnant Zell ewes (2.5 ± 1 years old, 42.19 ± 1.65 kg body weight [BW]) were selected 7 weeks before lambing. After receiving a ten-day adaptation diet, eight ewes were randomly allocated to each of the four treatments including: 1) The adaptation diet that fulfills ME and MP requirements, according to the National Research Council's (NRC, 2007) recommendations for late pregnant ewes (CON); 2) A diet with 10% higher levels of ME than CON, using corn grain for extra ME (HEC); 3) A diet with 10% higher level of ME compared to CON, using flaxseed for additional ME (HEF); and 4) A diet with a 10% higher level of MP than CON, using a rumen-protected lysin and methionine supplement (RP-Lys & Met) to supply extra MP (PRO). After lambing, ewes continued their dietary treatments until 30 days postpartum; however, the content of ingredients were proportionally increased to meet the recommended levels of energy and protein for lactating ewes (NRC, 2007). Blood samples from the ewes were taken on days -21, -14, and -7 before parturition. Colostrum yield and components were determined, and lamb's BW and milk intake (MI) were measured weekly until 28 days of age. Other growth performance indexes were computed accordingly. Blood samples from the offspring were taken on days 15 and 30 of life. The data was analyzed using SAS software (SAS Institute Inc., 9.4). Compared with the CON, HEF-fed ewes were the only group that had lower levels of oxidative stress and blood inflammatory markers at -7 d before parturition (P < 0.05). Concentrations of aminotransferases in CON-fed ewes were greater than other groups in late gestation (P ≤ 0.05). Offspring of HEF- and PRO-fed ewes had higher birth BW than lambs of the CON-fed group (3.71 and 4.11 vs. 3.01 ± 0.24, Kg for HEF, PRO, and CON, respectively). Except for week 2, lambs of CON-fed ewes had lower BW gain compared with the other offspring (P < 0.01). Lambs of PRO-fed ewes had greater levels of plasma immunoglobulin G (IgG, days 15 and 30 of life; P < 0.05) and MI (from week 2; P < 0.001) than other groups. In conclusion, regardless of the source, moderate overfeeding of ME/MP was more beneficial for the immune status of late pregnant dams and the offspring's performance than non-overfed ones.

Moderate over-feeding of different sources of metabolizable energy and protein improved gestational insulin resistance markers and maternal metabolic status of sheep around lambing.

The objective of this study was to determine the effect of moderate over-nutrition of energy (using flaxseed as a source of n-3 fatty acids) and protein (using rumen-protected lysine and methionine) around parturition on prepartal insulin resistance (IR) and periparturient metabolic status and performance of ewes. For this purpose, 32 apparently healthy ewes (2.5 ± 1 years old, 42.19 ± 1.65 kg body weight (BW)) were randomly selected out of 200 pregnant Zell ewes on d 50 before the expected lambing. After receiving a ten-day adaptation diet, eight ewes were randomly allocated to one of the following four dietary treatments: 1) the adaptation diet that meets the metabolizable energy (ME) and protein (MP) requirements of ewes during late pregnancy as recommended by National Research Council = NRC (2007) (Control = CON). 2) A diet with 10% higher levels of ME than NRC (2007) recommendations using corn grain as a source of extra ME (HEC). 3) A diet with 10% higher levels of ME compared to NRC (2007) recommendations using flaxseed for additional ME (HEF). 4) A diet with a 10% higher level of MP than NRC (2007) recommendations using rumen-protected lysine and methionine as a source of extra MP (PRO). While the dietary treatments did not affect animals' dry matter intake (DMI), the CON ewes lost more BW starting from d 10 to d 30 relative to parturition compared to others (P < 0.05). Milk protein and fat yield (g/d) were significantly higher for PRO and HEF compared to HEC and CON ewes (P < 0.05). Moderately over-fed ewes with MP and ME had an improved insulin sensitivity (IS) compared to CON ones. The source of additional ME also impacted IR indices because HEC showed an enhancement in IS than HEF animals (P < 0.05). A lower IS in CON ewes was parallel with higher concentrations of plasma urea at prepartum (P < 0.01), lower levels of plasma cholesterol, triglyceride (P < 0.01) and glucose post-partum (P < 0.05), and a tendency for higher levels of beta-hydroxybutyrate (BHB) and non-esterified fatty acids (NEFA) in CON ewes during both pre- and post-partum periods. There were negative correlations between plasma urea, BHB, and NEFA with IR markers on d 7 prepartum in line with these results. Comparing data, we could not find a significant change in metabolic status among ewes fed with either PRO or HEF/HEC. In conclusion, the inclusion of 10% extra MP and ME could improve late gestation maternal IS and the metabolic status of dams around lambing.

Effects of alternative feed additives to medicinal zinc oxide on productivity, diarrhoea incidence and gut development in weaned piglets.

The use of medicinal zinc oxide (ZnO) must be phased out by 2022, thus prompting an urgent need for alternative strategies to prevent diarrhoea in weaner piglets. The objectives of this study were to assess the impact on weaner piglet performance, diarrhoea incidence and gut development, when (1) dietary ZnO supplementation was substituted by alternative commercial products based on macroalgae, specific probiotics or synbiotics, or (2) dietary ZnO inclusion was reduced from 2500 to 1500 ppm. A total of 4680 DLY piglets (DanBred, Herlev, Denmark), weaned around 35 days of age, were randomly assigned according to sex and BW to six different dietary treatment groups. A basal diet was supplemented with no ZnO (NC = negative control), 2500 ppm ZnO (PC = positive control), 1500 ppm ZnO (RDZ = reduced dose of ZnO) or commercial macroalgae (OceanFeed™ Swine = OFS), probiotic Miya-Gold or synbiotic GærPlus products. The piglets entered and exited the weaner unit at ~7.0 and 30 kg BW, respectively. In-feed ZnO was provided the first 10 days post-weaning, while the alternative supplements were fed throughout the weaner period. As expected, the average daily feed intake, average daily weight gain (ADG), feed conversion ratio (FCR) and diarrhoea incidence were improved in the PC compared to NC group (P < 0.05) during phase 1 consistent with improved indices of villi development observed in subgroups of piglets sacrificed 11 days post-weaning. Reduction of ZnO to 1500 ppm lowered ADG (P < 0.05) and slightly increased incidence of diarrhoea during the first 10 days after weaning (but not later) without affecting FCR. None of the three alternative dietary additives, including a 10-fold increased dose of GærPlus than recommended, improved piglet performance, gut health and gut development above that of NC piglets. The OFS piglets sacrificed 11 days after weaning had significantly lower weights of hindgut tissue and contents compared to the PC group, consistent with antimicrobial activity of the product, which was detected from anaerobic in vitro fermentation. In conclusion, dietary ZnO supplementation during the first 10 days post-weaning may be reduced from 2500 to 1500 ppm without major negative implications for weaner piglet performance and health in herds under a high management level. However, none of the alternative dietary supplements were able to improve piglet performance or gut health, when ZnO was omitted from the diet.

Short communication: Relationship between body condition score and plasma adipokines in early-lactating Holstein dairy cows.

We hypothesized that plasma adipokine concentrations of early-lactation dairy cows are related to body condition score (BCS) at calving and to markers of metabolic status of the cow. As part of a larger study with 117 multiparous Holstein dairy cows, which had high BCS (BCS >4.0) or normal BCS (3.25-3.5) at calving, 22 cows were randomly selected (n = 11 per group) to be enrolled in this study. Cows were divided into 2 groups based on their BCS at calving: (1) normal BCS with BCS of 3.35 ± 0.13 (mean ± SD) and (2) high BCS cows with BCS of 4.14 ± 0.17. The 22 selected animals did not have a clinically diagnosed health problem after calving. Blood samples were taken right after calving (d 1) and before morning feeding on d 8, 15, and 21 postpartum concurrently with body condition scoring for all cows. Blood samples were analyzed for plasma adiponectin, leptin, tumor necrosis factor-α, and IL-6. The mean BCS remained highest in high-BCS cows during the first 21 d in milk. Leptin concentrations decreased progressively for all cows after calving. However, differences in BCS at calving were not related to leptin concentrations. Adiponectin, IL-6, and tumor necrosis factor-α concentrations were neither influenced by days in milk nor BCS after calving. Leptin and the leptin-to-adiponectin ratio did not show any correlation at any time point during the first 21 d in milk with plasma concentrations of nonesterified fatty acids or ß-hydroxybutyrate, which are considered as markers of metabolic status. Only for IL-6 at d 8 did we find a strong correlation with metabolic status indicators. In conclusion, plasma adipokine concentrations during the first 3 wk postpartum were not related to BCS in lactating Holstein cows that were clinically healthy at calving.

Effects of short-term feeding of different sources of fatty acids in pre-mating diets on reproductive performance and blood metabolites of fat-tailed Iranian Afshari ewes.

The effects of dietary omega-3 and omega-6 fatty acids (FAs) in pre-mating diets on reproductive performances and metabolic status of ewes have not been well investigated. Therefore, this study investigated the effect of supplementing different source of FAs on different aspects of reproductive performance of fat-tailed Iranian Afshari ewes. Thirty-two cycling, multiparous Afshari ewes were divided into four groups and fed one of the isocaloric and isonitrogenous diets supplemented with Sunflower oil (SFO), Fish oil (FO), calcium salt of palm oil (CaPO) and/or an equal mixture of SFO + FO (SFOFO). One day after the start of the flushing, the estrous cycles of all ewes were synchronized using 12-day CIDRs. After detecting signs of estrous, all ewes were mated with rams for 2 days. The examination of ovarian follicles was done by a transrectal ultrasonography. After counting all visible follicles, they were classified into 3 classes based on their diameters: small (≤3 mm), medium (3-4 mm) and large (≥4 mm). The results showed that fat sources had no impact on dry matter intake (DMI) and body condition score (BCS) of animals (P > 0.05). Plasma glucose, total cholesterol, and albumin concentrations also were not significantly affected by supplemental fat sources (P > 0.05). Plasma insulin levels were higher at the end of the flushing (on the day of CIDR removal) and the day of mating for ewes on CaPO diet (P < 0.05). Plasma estradiol-17ß concentration was greater on the day of mating for ewes fed FO (P < 0.05). Progesterone levels in pregnant ewes fed FO were higher 30 d after mating (11.02 ng/mL; P < 0.05). Dietary treatments had no significant effect on number of small, medium and total follicles as well as the average size of small and medium follicles at the end of the flushing and the day of mating. The number of medium follicles on the day of oestrus (mating) were significantly higher for the ewes fed FO or SFO diets (P < 0.05). Likewise, the mean number and size of large follicles at the end of flushing and the day of oestrus were higher in ewes on FO diet (P < 0.05). Lambing rate and twinning rate increased (P < 0.05) in ewes fed FO or SFO, respectively. In conclusion, supplementation of n-3 PUFA rich FA especially FO in pre-mating diets showed beneficial effects on some indices of reproductive performance of Afshari ewes including lambing and twining rate due to higher number of medium sized follicles and size of the ovulatory (large) follicles at the day of oestrus and increased plasma progesterone levels in pregnant ewes.