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.

The potential interaction between body condition score at calving and dietary starch content on productive and reproductive performance of early-lactating dairy cows.

Improving reproductive performance is one of the most important factors affecting the profitability of dairy herds. This study investigated the effect of feeding a high starch (HS) diet and body condition score (BCS) at calving on blood metabolites, fertility and ovarian function and milk production in Holstein dairy cows. One hundred seventy-four multiparous cows were fed common close-up and early lactation diets during the first 15 days in milk (DIM). Cows were randomly assigned to 1 of 2 experimental diets from 16 until 50 DIM (n = 87 per group); normal starch (228 g/kg diet DM; NS) or HS (270 g/kg diet DM; HS) diets. Each treatment group was further subdivided based on BCS at calving as normal BCS (BCS ⩽ 3.5; normal BCS (NBCS); n = 45) or high BCS (HBCS) (BCS ⩾ 3.75; HBCS; n = 42). A significant difference was detected for increased milk production (47.24 v. 44.55 kg/day) and decreased milk fat (33.93 v. 36.33 g/kg) in cows fed HS or NS, respectively. Plasma glucose and insulin concentrations were significantly higher in cows fed the HS compared to the NS diet. Diets significantly affected DIM at first artificial insemination (AI, 79.51 ± 3.83 v. 90.40 ± 3.83 days for cows fed HS and NS diets, respectively). High BCS groups had greater milk fat content and elevated plasma nonesterified fatty acids (NEFA), ß hydroxybutyrate (BHB) and bilirubin concentrations. In general, feeding higher starch diets to normal BCS cows during the first 50 DIM improved productive and reproductive performance of early-lactating dairy cows.

Cereals level and source effects on rumen fermentation, colostrum and milk properties, and blood metabolites in periparturient ewes.

Optimal type and dietary inclusion rates of cereal grains for periparturient sheep are unknown. The objective was to determine effects of feeding diets with high (H) v. low (L) levels of ground corn grain (CN) v. combined ground wheat and barley grains (WB) on intake, rumen fermentation, colostrum and milk properties, and blood metabolites of periparturient sheep. Twenty Afshari×Merino ewes were used in a completely randomized design study from 24 days prepartum through 21 days postpartum. Ewes were kept indoors in individual boxes and received once daily at 0900 h total mixed rations. Treatments were mixed rations containing either (1) H or (2) L concentrate based on either (1) 100% CN or (2) 50 : 50 ratio of ground wheat : ground barley grains in a 2×2 factorial arrangement. Each treatment group had five ewes including two twin-lamb ewes and three single-lamb ewes. Postpartal dry matter intake (DMI) increased by feeding high CN v. high and low WB, while high v. low CN improved postpartum DMI. The DMI during lambing tended to increase with the high v. low WB. Feeding CN v. WB, and feeding both CN and WB at L v. H level increased fecal pH. Postpartal rumen pH was lower with the high v. low WB (5.7 v. 6.2). Rumen concentrations of propionate were lower and of acetate were higher with L v. H grain levels. Increased dietary grain reduced urine pH for WB (7.24 v. 7.83) but not for CN (7.63 v. 7.52) prepartum. Colostrum properties, postpartal urine pH, lamb weight at birth and 21 days of age, and placental weight and expulsion time were unaffected. Milk yield increased and milk fat yield tended to increase by H v. L grain diets. Plasma glucose was increased by feeding high v. low WB, whereas CN v. WB tended to reduce peripartal plasma non-esterified fatty acids (NEFA) and increased insulin to NEFA ratio. In conclusion, more cereal grains can be included in periparturient sheep diets and CN instead of WB may be fed to periparturient sheep to improve energy status. Findings suggest opportunities to optimize periparturient ewe physiology and performance through feeding certain cereals and avoiding high levels of WB.

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.

Short communication: Effects of diets containing supplemental fats on ruminal fermentation and milk odd- and branched-chain fatty acids in dairy cows.

There is a growing interest in odd- and branched-chain fatty acids (OBCFA) in milk following reports that several branched-chain fatty acids (FA) have health promoting effects, and certain milk OBCFA could serve as a biomarker to assess ruminal function. Twenty-four Holstein cows were fed 3 low-forage diets containing 30 g/kg of dry matter of prilled palm fat (PPF), sunflower oil (SO), or an equal mixture of both fats (experiment 1) or 3 diets containing 30 g/kg of dry matter of SO with a forage-to-concentrate ratio of 39:61, 44:56, or 48:52 (Experiment 2); diets were fed to investigate milk OBCFA composition and to explore the relationships between ruminal VFA and milk OBCFA using principal component analysis. Including SO in diets decreased yields of milk 13:0 anteiso, 15:0 anteiso, 15:0, 17:0, cis-9 15:1, and cis-9 17:1 compared with PPF. The molar proportion of ruminal propionate was the lowest and the yields of milk 14:0 iso and 16:0 iso were the greatest with the diet containing both fat supplements. Replacing concentrate with forages linearly increased ruminal acetate and yields of milk 13:0 iso, 14:0 iso, 15:0 iso, 16:0 iso, 17:0 iso, 13:0 anteiso, 15:0 anteiso, 15:0, 17:0, cis-9 15:1, and cis-9 17:1. The principal component analysis revealed that ruminal molar proportion of acetate related to concentrations of milk iso FA containing <17-carbon, whereas ruminal propionate related to milk 15:0, 17:0, cis-9 15:1, and cis-9 17:1, with the stronger correlations between milk OBCFA and ruminal acetate than propionate. No associations were found between ruminal molar proportion of butyrate and milk OBCFA concentrations. The results suggest that complete replacement of PPF with SO at 30 g/kg of dry matter in low-forage diets is not an effective strategy to enhance bioactive branched-chain FA in milk, rather this feeding practice lowers anteiso FA in milk; however, increasing forage proportion in diets containing SO enhances several iso and anteiso FA in milk. The milk OBCFA concentrations have stronger correlations with ruminal acetate molar proportion than with propionate or butyrate in cows fed diets containing supplemental fats.

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.

Effects of heat stress on metabolism, digestibility, and rumen epithelial characteristics in growing Holstein calves.

To distinguish between the direct effects of heat stress (HS) and reduced DMI on metabolic and physiological variables, morphometry, and gene expression of transporters involving in ruminal VFA absorption, 16 Holstein bull calves (178.1 ± 7.55 kg of BW) were used in a randomized complete design using 2 experimental periods. In period 1 (P1), all animals were housed in thermoneutral (TN) conditions (20.7 ± 3.0°C and a temperature-humidity index [THI] of 65.2 ± 3.1) and fed ad libitum for 8 d. In period 2 (P2; 9 d), half of the calves ( = 8) were subjected to HS condition (29.9 to 41.0°C and a THI ≥ 85 for 8 h/d) and the other half ( = 8) were maintained in TN conditions but were pair fed (pair-fed thermoneutral [PFTN]) to the HS calves. Heat stress decreased DMI (20%; < 0.01) and, by design, PFTN calves had similar reduced intakes. During P1, ADG was similar between groups (1.7 kg/d), but ADG was reduced similarly for both groups (0.14 kg/d) during P2 ( < 0.01). Compared with P1, HS calves had decreased plasma glucose levels (12.5%; < 0.01) in P2. Basal insulin levels increased in P2 for HS calves (71%; < 0.03), but there was no difference between periods for PFTN calves. Circulating NEFA concentrations did not differ between periods in the HS calves, but PFTN animals had increased basal NEFA levels (100%; < 0.01). Compared with P1, HS calves had increased blood urea nitrogen concentration in P2 ( < 0.01) but blood urea nitrogen did not differ between periods in the PFTN calves. Heat stress had no measurable effect on nutrient digestibility. Rumen papillae height increased (51%; < 0.04) in HS calves whereas papillae top width decreased (40%; < 0.03), but there was no difference between treatments for other morphometric parameters. Gene expression of VFA transporters in rumen epithelium was not affected by thermal treatments. These results confirmed that short-term HS in the current study had no effect on diet digestibility, rumen morphology, or VFA transporters genes. Therefore, HS-induced decrease in DMI is in large part responsible for gross changes in BW in growing calves (albeit tissue composition of growth is not unknown).

The effect of cyclical and mild heat stress on productivity and metabolism in Afshari lambs.

To investigate the effect of heat stress (HS) on production and metabolism of Afshari sheep, 32 intact male lambs (33.2 ± 4.5 kg) were used in a completely randomized design using 2 experimental periods. In period 1 all 32 lambs were housed in thermal neutral (TN) conditions (25.6 ± 2.6°C and a temperature-humidity index [THI] of 72.0 ± 2.6) and fed ad libitum for 8 d. In period 2 (P2; 9 d), 16 lambs were subjected to cyclical HS (29.0 to 43.0°C and a THI ≥80 for 24 h/d) and the other 16 lambs were maintained in TN conditions but pair fed (pair-fed thermal neutral [PFTN]) to the HS lambs. During each period DMI and water intake were measured daily. Respiration rate, rectal temperature, and skin temperature at the shoulder, rump, and front and rear leg were recorded at 0700 and 1400 h daily. Water intake increased (P < 0.05) during P2 in both HS and TN lambs (88 and 35%, respectively). Heat stress increased the 0700 and 1400 h surface temperature at the shoulder (3.0 and 10.6%), rump (2.7 and 12.7%), rear leg (3.1 and 13%), and front leg (3.0 and 13%) and respiratory rates (72 and 124%; P < 0.01, respectively, for 0700 and 1400 h) but only the 1400 h rectal temperature was increased (P < 0.01; 0.54°C) in HS lambs. Plasma glucose concentration decreased in P2 (P < 0.01) in both the HS and PFTN lambs. Basal insulin concentrations decreased in PFTN controls but increased in HS lambs (environment × period interaction; P < 0.05). Blood urea nitrogen concentration was not affected by environment or period, but NEFA levels were slightly elevated (P < 0.01) in both PFTN and HS lambs during P2. Interestingly, HS did not affect DMI, but ADG was reduced (36%; P < 0.01) compared to the PFTN lambs. These results indicate that the direct effects of heat (not mediated by reduced DMI) are partially responsible for reduced growth in heat-stressed lambs.