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.

The effects of early- or mid-gestation nutrient restriction on bovine fetal pancreatic development.

Research on the effects of nutrient restriction in beef cows on fetal pancreatic development is limited. To address this, multiparous Angus-cross cows (n = 22) were fed either control (CON; to gain 1 kg/wk) or nutrient-restricted (NR; 0.55% NEm) diets based on NRC requirements. On d 30 of gestation, cows were blocked by body condition and randomly assigned to one of three nutritional regimes: CON fed from d 30 to 190 (n = 8), or NR/C (n = 7) or C/NR (n = 7) fed either the CON or NR diet from d 30 to 110 followed by CON or NR from d 110 to 190 of gestation. Cows were harvested on d 190 of gestation, and blood samples, fetal weights, and fetal tissue weights and samples were collected. Pancreas samples were embedded in paraffin and sectioned for standard immunohistochemistry procedures to quantify insulin-positive ß cells and number of apoptotic ß cells using TUNEL staining. Data were analyzed via ANOVA using the general linear model procedure of SAS. At harvest, empty carcass weights were decreased (P = 0.036) in fetuses of C/NR and NR/C fed dams compared to fetuses of CON fed dams. Pancreas weight was decreased (P = 0.028) in fetuses of C/NR fed dams compared to CON fetuses; however, fetuses of NR/C fed dams were not different (P > 0.05) from fetuses of CON fed dams. Maternal and fetal serum insulin concentrations were not different (P > 0.05) in NR/C fed compared to CON fed; however, concentrations of insulin were decreased (P = 0.036 and P = 0.40, respectively) in C/NR fed compared to CON fed. Beta cell number was decreased (P = 0.009) in fetuses of NR/C and C/NR fed dams compared to fetuses of CON fed dams. Percentage of apoptotic cells was increased (P < 0.0001) in fetuses of NR/C and C/NR fetuses than fetuses of CON fed dams. This evidence suggests that nutrient restriction either during early- or mid-gestation can negatively impact fetal pancreatic development. However, mid-gestational nutritional insult is potentially recovered by reacclimation to a diet that meets requirements of the dam, thus reducing negative outcomes in fetal offspring.

Bovine dopamine type-2 receptor SNP has no effect on growth, semen characteristics and prolactin concentrations in beef bulls treated with a dopamine agonist.

A dopamine type-2 receptor (DRD2) SNP, previously found to be correlated with serum prolactin (PRL) concentrations in cattle, was evaluated for impact on growth traits, serum prolactin concentration, and semen quality. Over a four-year period, yearling beef bulls were allowed diets containing or lacking ergot alkaloids (EA). Every 21 or 28 d semen was collected for semen motility and morphology assessment and blood samples were collected to measure serum PRL concentrations. In addition, body condition score and scrotal circumference were evaluated. Serum PRL concentrations were assessed using a radioimmunoassay. In the first year, all bulls were sacrificed at the end of a 126-day study. Testicles and epididymis were collected at the end of the study or 60 days after removal from treatment. Immunohistochemistry was performed on testis, epididymis, and sperm cells, incubated with or without a primary antibody for DRD2 and counterstained with DAPI. Isolation of DNA was performed on sperm pellets using DNAzol (Thermo Fisher Scientific, Waltham, MA, USA) methods. Polymerase chain reaction was performed to amplify the region of the DRD2 gene containing the SNP of interest. The products were subjected to restriction fragment length polymorphism analysis. Further, all samples were subjected to genotyping using a custom Taqman genotyping assay (Applied Biosystems, Foster city, CA, USA). The presence of DRD2 was detected in the testis, epididymis, and sperm cells. The DRD2 genotype was not associated with semen quality, serum PRL, or growth traits. Consumption of EA resulted in lesser PRL serum concentrations but had no effect on values for other variable examined.

Late-gestation protein restriction negatively impacts muscle growth and glucose regulation in steer progeny.

The objective of this study was to determine whether the amount of protein provided to cows during late gestation would affect postnatal growth and lead to changes in glucose and insulin concentrations. At 129 d of gestation, 10 mature multiparous Angus cows were stratified by body weight (BW) and body condition score (BCS) and allotted to either low protein level (LP, 6% crude protein [CP]) or high protein level (HP, 12% CP) groups. After calving, cows were managed together on improved pastures, which provided forage in excess of requirements until weaning. Male calves were maintained as a group after weaning on native range until 23 mo of age when individual steers were placed in single pens and fed a finishing diet for 84 d. The 12th rib fat thickness and longissimus muscle area were measured during finishing phase by ultrasound. Twenty days before the end of the finishing phase, steers were subjected to an intravenous glucose tolerance test. Steers were harvested and carcass characteristics collected. Cows' BW and BCS were similar at the initiation of treatment. During treatment HP dams gained 21 kg, whereas LP dams lost 7 kg (P = 0.04). Protein nutrition during late gestation did not influence calf birth weight, BW at weaning, adjusted 205 d BW, or average daily gain during lactation (P > 0.10). Longissimus muscle area measure by ultrasound was greater (P = 0.02) in HP steers at the beginning and end of finishing phase. Fat thickness of the 12th rib was not different (P > 0.10) between treatments. Glucose concentration after intravenous administration decreased (P = 0.002) in LP compared with HP steers. Peak of serum insulin concentration was greater (P = 0.04) and serum insulin concentration tended to decrease (P = 0.08) more rapidly in LP compared with HP steers after glucose infusion. At harvest, hot carcass weight was similar between treatments, but dressing percentage was increased (P = 0.05) in HP compared with LP steers. These data demonstrate that a lower protein nutrition level of dams during late gestation affect carcass characteristics and alter glucose regulation enhancing insulin secretion in steer progeny.

EGFR mutation subtypes and response to immune checkpoint blockade treatment in non-small-cell lung cancer.

BACKGROUND: Although EGFR mutant tumors exhibit low response rates to immune checkpoint blockade overall, some EGFR mutant tumors do respond to these therapies; however, there is a lack of understanding of the characteristics of EGFR mutant lung tumors responsive to immune checkpoint blockade. PATIENTS AND METHODS: We retrospectively analyzed de-identified clinical and molecular data on 171 cases of EGFR mutant lung tumors treated with immune checkpoint inhibitors from the Yale Cancer Center, Memorial Sloan Kettering Cancer Center, University of California Los Angeles, and Dana Farber Cancer Institute. A separate cohort of 383 EGFR mutant lung cancer cases with sequencing data available from the Yale Cancer Center, Memorial Sloan Kettering Cancer Center, and The Cancer Genome Atlas was compiled to assess the relationship between tumor mutation burden and specific EGFR alterations. RESULTS: Compared with 212 EGFR wild-type lung cancers, outcomes with programmed cell death 1 or programmed death-ligand 1 (PD-(L)1) blockade were worse in patients with lung tumors harboring alterations in exon 19 of EGFR (EGFRΔ19) but similar for EGFRL858R lung tumors. EGFRT790M status and PD-L1 expression did not impact response or survival outcomes to immune checkpoint blockade. PD-L1 expression was similar across EGFR alleles. Lung tumors with EGFRΔ19 alterations harbored a lower tumor mutation burden compared with EGFRL858R lung tumors despite similar smoking history. CONCLUSIONS: EGFR mutant tumors have generally low response to immune checkpoint inhibitors, but outcomes vary by allele. Understanding the heterogeneity of EGFR mutant tumors may be informative for establishing the benefits and uses of PD-(L)1 therapies for patients with this disease.

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.

Assessment of semen quality and fertility in young growing beef bulls exposed to ergot alkaloids.

The objective of this study was to evaluate semen quality and fertility in beef bulls grazing the ergot alkaloid (EA) producing tall fescue cultivar, Kentucky 31 (KY31), compared to a novel endophyte (NE) cultivar lacking EA. Two studies were conducted over a 3-year period. In studies 1 (n = 10; ages ≥ 24 mo) and 2 (n = 53 over two years; ages 12-16 mo), Angus (AN) bulls were stratified by body weight (BW), body condition score (BCS), and scrotal circumference (SC), and then allotted to graze either KY31 or NE for 56 days. Semen samples were collected, and BW, BCS, and SC were evaluated at the start of treatment (TRT) on day (d) 0 and every 28 days to the end of each study. In addition, blood samples were collected on d 0 and every 28 days for assessment of circulating prolactin (PRL) levels in study 2. On d 56, for both studies, semen from bulls (n = 2 per treatment in study 1 and n = 4 per treatment in study 2) with similar and acceptable quality were extended, kept at 19° C, and used for timed artificial insemination (TAI) of primi- and multiparous AN and AN- crossbred females. Pregnancy was evaluated at 35 and 90 days post-TAI via transrectal ultrasonography to determine pregnancy rates. Serum PRL concentrations showed a TRT by d effect (P ≤ 0.05), with values for bulls grazing KY31 decreased on d 28 and d 56 of grazing compared to NE. In studies 1 and 2, bull BW and BCS were affected by d (P ≤ 0.05), but not by TRT. No TRT or TRT by d effect on semen quality was observed in either study; however, d impacted both velocity and concentration in study 2 (P ≤ 0.05). In study 1, TAI pregnancy rates at 35 days post-TAI were lower (P ≤ 0.05) in the group inseminated with semen from bulls grazing KY31; however, in study 2, pregnancy rates did not differ due to treatment 35 post-TAI (P > 0.05). Grazing KY31 negatively impacted serum PRL concentrations, supporting previous observations; however, consumption of KY31 had no effect on growth or semen quality of AN bulls ranging from 12 to ≥24 mo of age. Furthermore, fertility data is inconsistent between studies and requires further investigation.

An evaluation of an immunomodulatory feed ingredient in heat-stressed lactating Holstein cows: Effects on hormonal, physiological, and production responses.

Holstein cows (n = 30) were balanced by days in milk, milk production, and parity (91 ± 5.9 d in milk, 36.2 ± 2.5 kg/d, and 3.1 ± 1.4, respectively) and fed OmniGen-AF (OG; Phibro Animal Health, Teaneck, NJ), an immune stimulant, at 0 g/cow per d for control (CON) or 56 g/cow per d for OG for 52 d on a commercial dairy. At 52 d of the study cows were randomly selected (n = 12) from both groups (6 OG and 6 CON) and housed in environmentally controlled rooms at the Agricultural Research Complex for 21 d at the University of Arizona. Cows were subjected to 7 d of thermoneutral (TN) conditions, 10 d of heat stress (HS), and 4 d of recovery (REC) under TN conditions. Feed intake, milk production, and milk composition were measured daily. Rectal temperatures (RT) and respiration rates (RR) were recorded 3 times per day (600, 1400, and 1800 h). Blood samples were taken on d 7 (TN), 8 (HS), 10 (HS), 17 (HS), and 18 (TN) during the Agricultural Research Complex segment. Cows in HS had higher RR and RT and water intake and lower dry matter intake and milk yield than these measures in TN. There was a treatment × environment interaction with cows fed OG having lower RR and RT and higher dry matter intake during peak thermal loads than CON. However, milk yield did not differ between groups. Cows fed OG had lower milk fat percent than CON (3.7 vs 4.3%) during HS. The SCC content of milk did not differ between treatment groups but rose in both groups during the REC phase following HS. Plasma insulin and plasma glucose levels were not different between groups. However, plasma insulin in both groups was lower during acute HS, then rose across the HS period, and was highest during the REC phase. Plasma cortisol levels were highest in all cows on the first day of HS (d 8) but were lower in cows fed OG compared with CON. However, plasma ACTH concentrations were elevated in OG-fed animals at all times samples were collected. Plasma ACTH was also elevated in cows fed both OG and CON during HS. Feeding OG reduced plasma cortisol during acute but not chronic HS and increased basal plasma ACTH, suggesting that OG treatment may alter the hypothalamic pituitary adrenal axis.

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.

Evaluation of dietary betaine in lactating Holstein cows subjected to heat stress.

Betaine (BET), a natural, organic osmolyte, improves cellular efficiency by acting as a chaperone, refolding denatured proteins. To test if dietary BET reduced the effect of heat stress (HS) in lactating dairy cows, multiparous, lactating Holstein cows (n=24) were blocked by days in milk (101.4±8.6 d) and randomly assigned to 1 of 3 daily intakes of dietary BET: the control (CON) group received no BET, mid intake (MID) received 57mg of BET/kg of body weight, and high dose (HI) received 114mg of BET/kg of body weight. Cows were fed twice daily and BET was top-dressed at each feeding. Cows were milked 2 times/d and milk samples were taken daily for analysis. Milk components, yield, feed intake, and water intake records were taken daily. Rectal temperature and respiration rate were taken 3 times/d at 0600, 1400, and 1800h. Cows were housed in environmentally controlled rooms and were allowed acclimation for 7d at thermoneutral (TN) conditions with a mean temperature-humidity index of 56.6. Cows were then exposed to 7d of TN followed by 7d of HS represented by a temperature-humidity index of 71.5 for 14d. This was followed by a recovery period of 3d at TN. Dietary BET increased milk yield during the TN period. No differences were found between BET and CON in total milk production or milk composition during HS. The increase in water intake during HS was not as great for cows fed BET compared with controls. The cows on CON diets had higher p.m. respiration rate than both MID and HI BET during HS, but lower rectal temperature compared with BET. No difference was found in serum glucose during TN, but cows given HI had elevated glucose levels during HS compared with CON. No differences were found in serum insulin levels between CON and BET but an intake by environment interaction was present with insulin increasing in HI-treated lactating dairy cows during HS. The heat shock response [heat shock protein (HSP) 27 and HSP70] was upregulated in bovine mammary epithelial cells in vitro. Blood leukocyte HSP27 was downregulated at the HI dose under TN conditions and HSP70 was upregulated at the HI dose and this effect was increased by HS. No effect was seen with the MID dose with HSP27 or HSP70. The lack of effect of BET at MID may be associated with uptake across the gut. We conclude that BET increased milk production under TN conditions and was associated with reduced feed and water intake and slightly increased body temperatures during HS of cows fed BET. The effect of BET on milk production was lost during HS with HI BET, whereas serum glucose levels increased during HS.

Modified homotopy perturbation method for solving hypersingular integral equations of the first kind.

Modified homotopy perturbation method (HPM) was used to solve the hypersingular integral equations (HSIEs) of the first kind on the interval [-1,1] with the assumption that the kernel of the hypersingular integral is constant on the diagonal of the domain. Existence of inverse of hypersingular integral operator leads to the convergence of HPM in certain cases. Modified HPM and its norm convergence are obtained in Hilbert space. Comparisons between modified HPM, standard HPM, Bernstein polynomials approach Mandal and Bhattacharya (Appl Math Comput 190:1707-1716, 2007), Chebyshev expansion method Mahiub et al. (Int J Pure Appl Math 69(3):265-274, 2011) and reproducing kernel Chen and Zhou (Appl Math Lett 24:636-641, 2011) are made by solving five examples. Theoretical and practical examples revealed that the modified HPM dominates the standard HPM and others. Finally, it is found that the modified HPM is exact, if the solution of the problem is a product of weights and polynomial functions. For rational solution the absolute error decreases very fast by increasing the number of collocation points.

Influence of excessive dietary protein intake during late gestation on drylot beef cow performance and progeny growth, carcass characteristics, and plasma glucose and insulin concentrations.

Spring-calving cows ( = 49) were used to investigate the effects of excessive prepartum dietary protein intake on late gestation cow performance as well as subsequent progeny growth, carcass characteristics, and plasma glucose and insulin concentrations. Treatments were formulated to be isocaloric and provide 100% (REQ) or 129% (HP) of CP requirement. Treatments were limit-fed 78 ± 12 d prepartum to calving. All cows were fed a common diet postpartum. Cow BW and BCS were recorded at initiation of treatments and within 48 h post-calving. Milk production was estimated via the weigh-suckle-weigh technique 69 ± 11 d postpartum. Calf BW was measured at birth and at weaning (121 ± 11 d of age). Progeny ( = 42) were weaned as a group and placed into a feedlot and fed a common finishing diet. Glucose and insulin concentrations were analyzed on a subset of progeny (12 per treatment) 90, 120, 150, 180, 210, and 240 min post-feeding, 2 d before slaughter (342 ± 11 d of age). Treatment had no effect ( ≥ 0.22) on cow BW, BCS, milk production, and subsequent reproduction or progeny preweaning growth. Progeny finishing growth and marbling scores were not affected ( ≥ 0.24) by treatment, yet 12th rib fat thickness ( < 0.01), KPH ( = 0.04), and YG ( = 0.01) were greater for progeny born to HP dams. Progeny born to HP dams had decreased ( ≤ 0.01) glucose and insulin concentrations, and insulin to glucose ratios, indicating greater insulin sensitivity. Although feeding cows 129% of CP requirement during late gestation did not affect cow performance or progeny preweaning or finishing period growth; carcass adiposity was increased by maternal treatment.

The effect of supplementing rumen undegradable unsaturated fatty acids on marbling in early-weaned steers.

The objective of this study was to determine if supplementation with a rumen undegradable unsaturated fatty acids (FA) source improved marbling deposition in early-weaned steers. All steers (Angus, = 23; Angus × Hereford, = 24) were weaned at 150 ± 5 d of age. Steers were blocked by BW and breed and then randomly assigned to either control (CON; average 1.5 kg of corn gluten feed [CGF], = 23) or isocaloric supplementation containing a rumen undegradable fat source (RUF; 200 g of Megalac-R added to an average of 1.06 kg of CGF, = 24) offered 5 d/wk for 110 d. All steers had ad libitum access to pastures throughout treatment and received supplements individually. Steer BW and blood samples were collected at 0, 55, and 110 d of supplementation, and real-time ultrasound measurements were collected at d 110. Following treatment, steers were transported to Oklahoma State University for finishing and subsequent harvesting at a commercial plant. All data were analyzed using the PROC MIXED procedure of SAS either as repeated measures or ANOVA depending on parameters. There were no significant changes in BW from the beginning of treatment to harvest due to treatment. Ultrasound data showed that RUF steers tended ( = 0.08) to have more i.m. fat than CON at d 110. Serum concentrations of FA showed a treatment × day interaction ( < 0.02) for 16:0, 18:0, 18:1-9, 18:2, 20:4, and total FA. These specific FA concentrations slightly increased in CON steers, but there was a more pronounced increase in the concentration of these FA across the supplementation period in RUF steers. Serum triglyceride and cholesterol concentrations were increased ( < 0.01) on d 55 and 110 in RUF steers compared with those in CON steers. Serum leptin concentration in RUF steers was greater ( < 0.01) than CON steers at d 110. After harvest, RUF carcasses had greater ( = 0.01) marbling scores than those of CON carcasses. All other carcass measures were similar between treatments. The percentage of total lipids was increased ( = 0.011) in steaks from RUF compared to CON. There was a decrease ( < 0.05) in adipocyte diameter in i.m. fat depot of RUF steers compared to CON. There was also a tendency ( = 0.06) for RUF steers to have a greater percentage of 20 to 30 µm adipocytes in their i.m. depot than CON steers. This study indicates that supplementation of unsaturated FA can positively impact marbling deposition in early-weaned steers without impacting other carcass measures.

Bulls grazing Kentucky 31 tall fescue exhibit impaired growth, semen quality, and decreased semen freezing potential.

Serum prolactin (PRL) and testosterone concentrations, body weight, body composition, semen quality, and semen freezing potential for bulls grazing the toxic tall fescue (Lolium arundinaceum [Schreb.] Darbysh. = Schedonorous arundinaceum [Schreb.] Dumort.) cultivar Kentucky 31 (E+) compared with a novel endophyte cultivar lacking ergot alkaloids (E-) were evaluated. Angus bulls were allotted to treatment (Day 0) and grazed E+ or E- for 155 days. Treatment-by-day interaction was significant (P < 0.05) for serum PRL concentrations with E+treated bulls exhibiting reduced PRL values compared with E- control bulls, but no differences were observed for serum testosterone concentrations (P > 0.05). Further, bulls on the E+ treatment exhibited decreased total gain, average daily gain, and body weight by Day 140 (P < 0.05) compared with the E- bulls. Rump muscle depth was lower because the treatment in bulls grazing E+ compared with E- (P < 0.05) and intramuscular fat in the E- bulls compared with the E+ group was higher by Day 155 (P < 0.05). Analysis of ejaculates showed significant treatment × day effects for sperm concentration with lower values observed for bulls on the E+ treatment (P < 0.05). The percent normal morphology was reduced in ejaculates from E+ bulls compared with E- bulls (P < 0.05), and the difference was due to an increase in abnormal sperm present in the E+ ejaculates from Day 84 to 140 (P < 0.05). In addition, spermatozoa motility and progressive motility were decreased on thawing in semen samples from E+ bulls compared with E- bulls (P < 0.05).