Effects of maternal nutrient restriction during gestation on bovine serum microRNA abundance.

The objective was to determine the effects of maternal nutrient restriction during gestation on serum microRNA (miRNA) abundance in cattle. Primiparous Angus-cross cows (n=22) were fed either control (CON; to gain 1 Kg/week) or nutrient restricted (NR; 0.55% NEm) diets based on National Research Council requirements. On day 30 of gestation, cows were blocked by body condition and randomly assigned to one of three diets: CON (n=8) days 30-190; NR (n=7) days 30-110 followed by CON days 110-190 (NR/C); or CON (n=7) days 30-110 followed by NR days 110-190 (C/NR). At 190 days of gestation, maternal serum was collected for RNA isolation and analyzed using a miRNA microarray of known Bos taurus sequences. Data were normalized using LOWESS and analyzed via ANOVA. At 190 days of gestation, 16 miRNAs exhibited differential abundance (P<0.05) between treatments. Cows that underwent NR, irrespective of when the insult occurred, had downregulated bta-miR-126-3p compared to CON cows. Bta-miR-16b was downregulated and three miRNAs upregulated in NR/C compared to C/NR and CON cows. Additionally, seven miRNAs were downregulated and four miRNAs upregulated in C/NR compared to NR/C and CON cows. Comparison of NR/C and C/NR cows revealed three differentially abundant (P<0.04) miRNAs (bta-miR-2487_L-2R-3_1ss15CT, bta-miR-215, and bta-miR-760-5p). Top KEGG pathway enrichment of target genes included: pathways in cancer, PI3K-Akt signaling, focal adhesion, Ras signaling, proteoglycans in cancer, and MAPK signaling. In summary, maternal nutrient restriction altered serum miRNA abundance profiles irrespective of the time at which the nutritional insult was induced.

Effects of nutrient restriction during early or mid-gestation in bovine on placental development and miRNA expression in the cotyledon.

The objective of this study was to determine effects of maternal nutrient restriction (NR) during early or mid-gestation on uterine composition and miRNA expression in cotyledons. Primiparous Angus-cross cows (n = 38) were synchronized and inseminated using male sexed semen, blocked by body condition score and body weight (BW), and assigned to treatments. Animals were fed either: control (CON; gain 1 kg/week) or NR (55% maintenance energy and crude protein requirements) based on BW. An initial set of animals were fed either NR (n = 8) or CON (n = 8) from day 30-110 of gestation. A second set of animals were fed CON (n = 8) d 30-190 (CON/CON); NR (n = 7) day 30-110 followed by CON day 110-190 (NR/CON); or CON (n = 7) day 30-110 followed by NR day 110-190 (CON/NR). Cows were harvested on day 110 or 190 of gestation to collect placental tissues. RNA was isolated from cotyledon samples (3 animals/group) prior to microarray analysis using known Bos taurus microRNA sequences. Relative microRNA abundance was analyzed via ANOVA. Maternal NR increased (P < 0.05) cotyledon weight and total placentome surface area irrespective of gestational day. At day 110 of gestation, 51 microRNAs were reduced while 91 microRNAs observed greater abundance (P < 0.05) in NR verses CON cotyledons. At day 190 of gestation, 40 microRNAs were reduced and 26 microRNAs were increased (P < 0.05) in both NR/CON and CON/NR verses CON cotyledons. Top KEGG pathway analysis included: axon guidance, endocytosis, neuroactive ligand receptor interaction, and MAPK signaling pathway. Early-gestation maternal NR altered microRNA abundance to a greater extent than mid-gestation NR.

Effects of post-weaning supplementation of immunomodulatory feed ingredient on body weight and cortisol concentrations in program-fed beef heifers.

The objective of this study was to determine the effects of an immunomodulatory feed ingredient during post-weaning on growth and cortisol concentrations of beef heifers. Commercial Angus heifers (n = 72) from 2 AI sires were blocked (n = 9) by BW and randomly assigned to one of 2 pens per block. Each pen (4 heifers/pen) per block was randomly assigned to either one of 2 treatments, Celmanax (CEL) or corn germ (CON). The heifers were fed a commercial total mixed ration (TMR) twice daily from d 0 to 60 to gain 0.75 kg/d. The feed was top-dressed once daily with either 72g of Celmanax (CEL) or corn germ (CON) per pen. Body weight (BW) was collected on d -1, 0, 15, 30, 45, 60, and 61. Blood samples were collected on d 0, 15, 30, 45, and 60. Two heifers per pen (n = 32) were randomly selected for a transportation challenge to evaluate stress response on d 62 or 63 of the study. Sixteen heifers (n = 8 CEL; n = 8 CON) were randomly selected for a corticotropin-releasing hormone/arginine vasopressin (CRH/AVP) challenge and an intravenous glucose tolerance test (IVGTT) on d 64 and 67 of the study. The pen was the experimental unit, and data were analyzed by ANOVA or repeated measures analysis as appropriate. Feed efficiency and BW gain were increased (P = 0.04) in CEL heifers compared to CON heifers. Serum cortisol concentrations were decreased (P < 0.01) in CEL heifers compared to CON heifers on d 30 to 60 post-weaning. Serum cortisol concentrations were decreased (P < 0.05) in CEL heifers compared to CON heifers throughout the transportation challenge. Serum cortisol concentrations were decreased (P < 0.05) in CEL heifers compared to CON heifers during the CRH/AVP challenge from 60 to 150 min post-infusion. Treatment did not influence (P = 0.29) plasma insulin or glucose (P = 0.63) concentrations during the IVGTT. In summary, supplementation of Celmanax post-weaning increased BW gain and reduced cortisol concentrations in challenged beef heifers.

Short communication: Manipulation of neonatal leptin profile via exogenous glucocorticoids in beef calves.

The link between circulating glucocorticoids and leptin in beef calves has not been explored but has been noted in several studies. The aim of this study is to determine the effects of exogenous glucocorticoids given at birth and 1 day of age on serum leptin concentrations in beef calves. Ruminant animals secrete leptin, which is thought to be important for the programming of the hypothalamic appetite centers. Angus crossbred cows (n = 31) bred via natural service were utilized for this experiment. At parturition (day 0), calf BW was recorded and each calf was infused intravenously with either a hydrocortisol sodium succinate solution (HC, 8 males and 8 females) at a dosage of 3.5 µg/kg of BW or a similar volume of saline solution (CONT, 7 males and 8 females). Each calf was given a second infusion of its respective treatment 24 h postpartum at 1.5 µg/kg of BW for HC treatment. Calf treatment was blocked by sex, dam body condition score (BCS), and dam age. Blood samples were taken via jugular venipuncture before infusion, daily from days 0 to 5, then every other day up to day 17. Serum leptin and cortisol concentrations were analyzed via radioimmunoassay. Dam age, dam BCS, calf BW, and serum leptin and cortisol concentrations were analyzed using MIXED procedure of SAS. Dam age was not different (P = 0.81) among HC and CONT calves (4.9±0.5 and 4.7±0.5, respectively). Dam BCS was not different between treatments (5.7±0.2 and 5.6±0.2 HC and CONT, respectively; P = 0.66). There was no difference in calf birth BW between treatments (P = 0.87) and averaged 38.3±1.4 kg. Cortisol concentrations were not different between both treatments (P = 0.23) from birth to day 4 of age. Calves that received the HC treatment showed significantly reduced (P = 0.03) leptin concentrations on days 1 to 13. Calf BW from 60 to 150 days of age was not different between CONT and HC treated calves (P = 0.65). These data indicate that exogenous glucocorticoids can be used to suppress neonatal leptin levels in calves. This could lead to changes in voluntary feed intake of treated calves.

The effects of late gestation nutrient restriction of dams on beef heifer intake, metabolites and hormones during an ad libitum feeding trial.

This study's objective was to determine if nutrient restriction during late gestation affected beef heifer feed intake, body weight (BW) gain and endocrine regulation during a 10-week feeding trial. During the last 100 days of gestation, control (CON) dams were fed to increase body condition score (BCS). Whereas, nutrient-restricted dams (NR) and NR dams protein supplemented 3 days/week (NRS) were fed to decrease BCS by 1.2. After parturition, all cow-calf pairs were moved to a common pasture and fed in excess of requirements until weaning. At 15 months of age, heifers were randomly sorted into two pens and adjusted to a commercial total mixed ration over a 2-week period. Blood samples and BW were taken at the initiation of feeding and on a biweekly basis for the duration of the feeding trial. Feed intake was monitored for 10 weeks using a GrowSafe System. After 10 weeks, an intravenous glucose tolerance test (IVGTT) was performed on 21 randomly subsampled heifers. During the feeding trial, NR heifers consumed more feed than CON and NRS heifers. Heifers from NR dams tended to increase BW compared to NRS and CON heifers when adjusted for initial BW. Heifers from NR and NRS dams had a greater increase in BCS compared to heifers from CON dams. Plasma glucose and insulin concentrations during the feeding trial increased in NR heifers compared to the other groups beginning at 2 and 4 weeks respectively. Plasma leptin concentrations were increased in the NR and NRS heifers compared to the CON heifers beginning at week 4 of feeding. During the IVGTT at the conclusion of the feeding challenge, plasma glucose and insulin were increased in NR heifers compared to other treatment groups. These results show that nutrient restriction during late gestation alters appetite and endocrine regulation in heifer offspring.

Effects of maternal nutrient restriction during early or mid-gestation without realimentation on maternal physiology and foetal growth and development in beef cattle.

The aim of this study is to determine the effects of early and mid-gestation nutrient restriction on maternal metabolites and foetal growth. Primiparous Angus cows were synchronized and inseminated with semen from one sire. Dietary treatments were: control to gain 1 kg/week (CON) or 0.55% maintenance energy and CP requirements (nutrient restricted; NR). A subset of dams was fed NR (n=8) or CON (n=8) from days 30 to 110 of gestation. Another group was fed CON (n=8), days 30 to 190; NR (n=7), days 30 to 110 followed by CON days 110 to 190; or CON, (n=7) days 30 to 110 followed by NR days 110 to 190. Cows were harvested at days 110 or 190 of gestation, when foetal measurements and samples were collected. Cows that were NR during days 30 to 110 or 110 to 190 of gestation lost significant BW and body condition score (P<0.001), this was associated with reduced plasma glucose during NR (P<0.002). Foetal weights, empty foetal weights, abdominal and thoracic circumferences were all reduced (P<0.03) in day 110 NR animals. Foetal perirenal adipose as a percentage of empty foetal weight was increased (P=0.01) in NR day 110 female foetuses compared with CON foetus. Maternal serum triglycerides at day 110 of gestation were decreased (P<0.05) in NR dams, whereas foetal serum triglycerides were increased (P<0.05) in response to maternal NR. Foetal weights tended to be reduced (P=0.08) in NR/CON and CON/NR v. CON/CON cattle at day 190 of gestation. Empty foetal weights, abdominal and thoracic circumferences were reduced (P⩽0.03) in NR/CON and CON/NR v. CON/CON cattle. Brain weight as a percentage of empty foetal weight was increased (P<0.001) in NR/CON and CON/NR v. CON/CON cattle. Foetal perirenal adipose as a percentage of empty foetal weight was increased (P=0.003) in NR/CON and CON/NR v. CON/CON cattle. Maternal serum triglycerides at day 190 of gestation were decreased (P<0.05) in association with maternal NR. Foetal serum triglycerides at day 190 of gestation were increased (P<0.05) in response to maternal NR during early gestation but decreased by NR in mid gestation compared with CON foetuses. The data show that maternal nutrient restriction during early or mid-gestation cause's asymmetrical foetal growth restriction, regardless if the restriction is preceded or followed by a period of non-restriction.

The effect of frequency of supplementing rumen-protected unsaturated fatty acids on blood serum fatty acid profiles in beef heifers and lactating cows.

The objective of this study was to determine if supplementation frequency of rumen-protected fat (RPF) influences circulating serum concentrations of fatty acids (FA), NEFA, and urea nitrogen in beef heifers and lactating cows. In Exp. 1, 12 early gestation beef heifers were supplemented 0.5 kg of corn gluten feed (CGF) daily during a 2-wk adaptation period. During the last 3 d of adaptation, blood samples were collected immediately before supplementation and then 8 and 16 h postsupplementation daily. Each heifer was then randomly assigned to 1 of 3 supplementation frequency treatments of RPF (3, 5, or 7 d/wk) for 3 wk in a 3 × 3 Latin square design with 3 periods (4 heifers per treatment per period), with each treatment receiving the same amount of RPF and CGF per wk (1.0 and 2.7 kg as fed, respectively). Blood samples were collected during the final 3 d of each supplementation period as described in the adaptation period. In Exp. 2, 18 Angus crossbred cows in early lactation were supplemented with 4.54 kg (as fed) of CGF weekly either at 3, 5, or 7 d/wk during a 2-wk adaptation period. Blood samples were collected during the last 3 d of adaptation as in Exp. 1. For the subsequent 3 wk, RPF (530, 318, and 227 g/d when supplemented) was added to the CGF supplement so that each supplementation frequency received 1.59 kg as fed/wk of RPF. Blood samples were collected during the last 3 d of supplementation as in Exp. 1. Serum FA profiles on a random subsample of 9 heifers in Exp. 1 and all animals in Exp. 2 were determined via gas chromatograph (GC), and values were analyzed using the MIXED procedure of SAS. In Exp. 1, there were no differences ( ≥ 0.53) in serum FA profile across supplementation frequencies. There was a decrease in serum 18:2, 18:1 -9 and total FA during the sampling time (time effect, < 0.02). In Exp. 2, there was treatment × time effect ( ≤ 0.001) for both 18:2 and total FA measured. The 7 d/wk frequency had a greater concentration of C18:2 and total FA at 8 h of the sampling period when compared to the other 2 frequencies. These results demonstrate that supplementation of RPF at 3, 5 or 7 d/wk resulted in no changes in serum FA profiles in growing heifers. In early lactating beef cows, there were minor time point changes in serum FA due to supplementation frequency; however, the vast majority of time points were not different, indicating no substantial changes in serum FA profiles.

The effects of late gestation maternal nutrient restriction with or without protein supplementation on endocrine regulation of newborn and postnatal beef calves.

A study was conducted to evaluate late gestation maternal nutrient restriction (NR) with or without protein supplementation on endocrine regulation in newborn beef calves. This study used multiparous cows (4 and 5 years of age, n = 57) randomly assigned to one of three treatments for the last 100 days of gestation. The control (Con; n = 19) cows were fed to increase body condition score, whereas the NR (n = 19) and NR with protein supplement (NRS, n = 19) cows were fed to lose 1.2 ± 0.2 body condition score units during the last 100 days of gestation. Control cows were allowed ad libitum access to tall fescue/crabgrass paddock and, when grazing became insufficient, ad libitum hay was provided along with 1.3 kg of corn gluten feed 5 days/wk. Tall fescue paddocks were strip grazed to limit forage availability for NR and NRS. The NRS-treated dams were individually penned and fed 0.45 kg of soybean meal 3 days/wk. As forage became limited, the nutrient-restricted paddocks received limited fescue hay. After parturition cow/calf pairs were moved to a common pasture and received ad libitum silage and high-concentrate feed. Maternal NR regardless of supplementation reduced cow plasma glucose and insulin concentrations during late gestation (P < 0.0001 and P = 0.0051, respectively). Calves from NR dams weighed less at birth than Con calves (P = 0.04), whereas NRS calves were intermediate (33.4 ± 1.2, 35.0 ± 1.3, and 37.2 ± 1.3 kg NR, NRS, and Con, respectively). Plasma glucose concentrations of unsuckled calves at birth were reduced (P = 0.037) in NR and NRS calves compared with Con (67.7 ± 6.5 and 60.1 ± 6.9 vs. 83.7 ± 6.1 mg/dL, respectively). At birth, Con and NRS calves had increased (P = 0.0037) plasma leptin concentrations compared with NR calves, whereas calf plasma cortisol concentrations were greater for the nutrient-restricted groups than the Con group (treatment × day P = 0.0135). Plasma IgG concentrations from calves at 5 days of age were similar (P = 0.701) between maternal late gestation treatments. This research reports that late gestation NR reduces postnatal calf birth weight, plasma glucose and leads to reduced plasma leptin. Maternal protein supplementation appears to partially alleviate the effects of late gestation NR on reducing plasma leptin, birth weight, and growth rate from Day 30 of age to weaning.

The natural history of pelvic organ support in pregnancy.

Little is known about the anatomic and physiologic changes in the pelvic floor that occur during pregnancy. The purpose of this study was to prospectively document pelvic organ support throughout pregnancy using the standardized system of the International Continence Society, also known as the Pelvic Organ Prolapse Quantification (POPQ) Staging System. Pelvic organ support evaluations were performed in nulliparous pregnant women presenting for routine obstetric care during each trimester. POPQ stage assignments and POPQ component measurements were compared for first-, second- and third-trimester examinations. Overall POPQ stage was significantly higher in the third trimester than in the first (P=0.001). Individual POPQ points which showed significant differences between the first and third trimesters include Aa, PB, Ap, Ba, Bp, TVL and GH. These findings probably represent normal physiologic changes of the pelvic floor during pregnancy, but suggest that significant changes may be objectively demonstrated prior to delivery.