Effects of excess metabolizable protein on ovarian function and circulating amino acids of beef cows: 1. Excessive supply from corn gluten meal or soybean meal.

In the dairy industry, excess dietary CP is consistently correlated with decreased conception rates. However, the source from which excess CP is derived and how it affects reproductive function in beef cattle is largely undefined. The objective of this experiment was to determine the effects of feeding excess metabolizable protein (MP) from feedstuffs differing in rumen degradability on ovulatory follicular dynamics, subsequent corpus luteum (CL) development, steroid hormone production and circulating amino acids (AA) in beef cows. Non-pregnant, non-lactating mature beef cows (n=18) were assigned to 1 of 2 isonitrogenous diets (150% of MP requirements) designed to maintain similar BW and body condition score (BCS) between treatments. Diets consisted of ad libitum corn stalks supplemented with corn gluten meal (moderate rumen undegradable protein (RUP); CGM) or soybean meal (low RUP; SBM). After a 20-day supplement adaptation period, cows were synchronized for ovulation. After 10 days of synchronization, gonadotropin releasing hormone (GnRH) was administered to reset ovarian follicular growth. Starting at GnRH administration and daily thereafter until spontaneous ovulation, transrectal ultrasonography was used to diagram ovarian follicular growth, and blood samples were collected for hormone, metabolite and AA analyses. After 7 days of visual detection of estrus, CL size was determined via ultrasound. Data were analyzed using the MIXED procedures of SAS. As designed, cow BW and BCS were not different (P⩾0.33). Ovulatory follicular wavelength, antral follicle count, ovulatory follicle size at dominance and duration of dominance were not different (P>0.13) between treatments. Cows supplemented with CGM had greater post-dominance ovulatory follicle growth, larger dominant follicles at spontaneous luteolysis, shorter proestrus, and larger ovulatory follicles (P⩽0.03) than SBM cows. No differences (P⩾0.44) in peak estradiol, ratio of estradiol to ovulatory follicle volume, or plasma urea nitrogen were observed. While CL volume and the ratio of progesterone to CL volume were not affected by treatment (P⩾0.24), CGM treated cows tended to have decreased (P=0.07) circulating progesterone 7 days post-estrus compared with SBM cows. Although total circulating plasma AA concentration did not differ (P=0.70) between treatments, CGM cows had greater phenylalanine (P=0.03) and tended to have greater leucine concentrations (P=0.07) than SBM cows. In summary, these data illustrate that excess MP when supplemented to cows consuming a low quality forage may differentially impact ovarian function depending on ruminal degradability of the protein source.

Effects of excess metabolizable protein on ovarian function and circulating amino acids of beef cows: 2. Excessive supply in varying concentrations from corn gluten meal.

In the dairy industry, excess dietary CP is consistently correlated with decreased conception rates. However, amount of excess CP effects on reproductive function in beef cattle is largely undefined. The objective of this experiment was to determine the effects of excess metabolizable protein (MP) supplementation from a moderately abundant rumen undegradable protein (RUP) source (corn gluten meal: 62% RUP) on ovarian function and circulating amino acid (AA) concentrations in beef cows consuming low quality forage. Non-pregnant, non-lactating beef cows (n=16) were allocated by age, BW and body condition score (BCS) to 1 of 2 isocaloric supplements designed to maintain BW for 60 days. Cows had ad libitum access to corn stalks and were individually offered a corn gluten meal-based supplement daily at 125% (MP125) or 150% (MP150) of National Research Council (NRC) MP requirements. After a 20-day supplement adaptation period, cows were synchronized for ovulation. After 10 days of synchronization, follicular growth was reset with gonadotropin releasing hormone. Daily thereafter, transrectal ultrasonography was performed to diagram ovarian follicular waves, and blood samples were collected for hormone, metabolite and AA analyses. After 7 days of observation of estrus, corpus luteum (CL) size was determined via ultrasound. Data were analyzed using the MIXED procedures of SAS. No differences (P⩾0.21) in BW and BCS existed throughout the study; however, plasma urea N at ovulation was greater (P=0.04) in MP150. Preovulatory ovarian follicle size at dominance, duration of dominance, size at spontaneous luteolysis, length of proestrus and wavelength were not different (P⩾0.11) between treatments. However, ovulatory follicles were larger (P=0.04) and average antral follicle count was greater (P=0.01) in MP150 than MP125. Estradiol concentration and ratio of estradiol to ovulatory follicle volume were not different due to treatment (P⩾0.25). While CL volume 7 days post-estrus was greater (P<0.01) in MP150 than MP125, circulating progesterone 7 days post-estrus and ratio of progesterone to CL volume were not different (P⩾0.21). Total AA were not different (P⩾0.76) at study initiation or completion; however, as a percent of total AA, branched-chain AA at ovulation were greater (P=0.02) in MP150. In conclusion, supplementation of CP at 150% of NRC MP requirements from a moderately undegradable protein source may enhance growth of the ovulatory follicle and subsequent CL compared with MP supplementation at 125% of NRC MP requirements.

Effects of supplementing calcium salts of polyunsaturated fatty acids to late-gestating beef cows on performance and physiological responses of the offspring.

This experiment compared performance and physiological responses of the offspring from cows supplemented with Ca salts of PUFA or SFA + MUFA during late gestation. Ninety-six multiparous, nonlactating, pregnant Angus × Hereford cows were ranked by BW, BCS, and age and divided into 24 groups of 4 cows/group at the end of their second trimester of gestation (d -7). Cows conceived during the same estrus synchronization + AI protocol, with semen from a single sire; hence, gestation length was 195 d for all cows at the beginning of the experiment (d 0). Groups were randomly assigned to receive (DM basis) 405 g/cow daily of soybean meal in addition to 1) 190 g/cow daily of Ca salts of PUFA based on eicosapentaenoic, docosahexaenoic, and linoleic acids or 2) 190 g/cow daily of Ca salts of SFA + MUFA based on palmitic and oleic acids (CON). Groups were maintained in 2 pastures (6 groups of each treatment/pasture) and received daily 10.1 kg/cow (DM basis) of grass-alfalfa hay. Groups were segregated into 1 of 12 drylot pens (6 by 18 m) and individually offered treatments 3 times/wk from d 0 until calving. Cow BW and BCS were recorded, and blood samples were collected on d -7 of the experiment and also within 12 h after calving. Calf BW was also recorded within 12 h of calving. Calves were weaned on d 280 of the experiment, preconditioned for 45 d (d 280 to 325), transferred to a growing lot on d 325, and moved to a finishing lot on d 445, where they remained until slaughter. At calving, PUFA-supplemented cows had a greater ( < 0.01) proportion (as % of total plasma fatty acids) of PUFA, including linoleic, linolenic, arachidonic, docosapentaenoic, and docosahexaenoic acids. At weaning, calves from CON-supplemented cows were older ( = 0.03), although no treatment differences were detected ( = 0.82) for calf weaning BW. During both growing and finishing phases, ADG was greater ( ≤ 0.06) in calves from PUFA-supplemented cows. Upon slaughter, HCW and marbling were also greater ( ≤ 0.05) in calves from PUFA-supplemented cows. Collectively, these results indicate that supplementing eicosapentaenoic, docosahexaenoic, and linoleic acids to late-gestating beef cows stimulated programming effects on postnatal offspring growth and carcass quality. Therefore, supplementing late-gestating beef cows with Ca salts of PUFA appears to optimize offspring productivity in beef production systems.

Supplementation of metabolizable protein during late gestation and fetal number impact ewe organ mass, maternal serum hormone and metabolite concentrations, and conceptus measurements.

To examine the effects of maternal metabolizable protein (MP) supplementation during late gestation on serum hormone and metabolites and organ masses, multiparous ewes (n = 45) carrying singletons or twins were allotted randomly (within pregnancy group) to 1 of 3 treatments: 60% (MP60), 80% (MP80), or 100% (MP100) of MP requirements. Blood samples were drawn before the initiation of diets (day 100) and before slaughter (day 130) for chemistry panel analysis and weekly for hormone analysis including progesterone (P4) and estradiol-17ß (E2). At day 130, ewe organ masses were recorded. Despite being fed isocaloric diets, MP60 ewes gained less weight throughout pregnancy compared with MP80 and MP100 ewes which were similar. Although diet did not impact E2 or P4 concentrations, ewes carrying twins had greater (P < 0.05) concentrations of both as gestation advanced. Albumin, aspartate aminotransferase, and total protein were reduced (P < 0.05) in MP60 compared with MP100 ewes near term. There was a diet by fetal number interaction (P = 0.03) for lactate dehydrogenase. Twin-carrying MP80 ewes had greater lactate dehydrogenase compared with all other groups on day 130 of gestation. Ewes that were fed MP80 had greater body weight on day 130 of gestation compared with MP60 ewes. Kidney and heart weights were lighter in MP60 ewes compared with MP80 ewes. There was a maternal diet by fetal number interaction (P = 0.05) on fetal weight per unit empty ewe body weight. In ewes carrying singletons, MP60 ewes supported less fetal weight compared with MP100. In contrast, MP60 ewes supported more fetal mass compared with MP100 ewes when carrying twins. The level of protein, and not just total energy, in the diet appears to impact some aspects of the maternal system. Moreover, it appears some measurements of mobilizing maternal body resources are enhanced in ewes carrying twins.

Effects of protein supplementation frequency on physiological responses associated with reproduction in beef cows.

The objective of this experiment was to determine if frequency of protein supplementation impacts physiological responses associated with reproduction in beef cows. Fourteen nonpregnant, nonlactating beef cows were ranked by age and BW and allocated to 3 groups. Groups were assigned to a 3 × 3 Latin square design, containing 3 periods of 21 d and the following treatments: 1) soybean meal supplementation daily (D), 2) soybean meal supplementation 3 times/week (3WK), and 3) soybean meal supplementation once/week (1WK). Within each period, cows were assigned to an estrus synchronization protocol: 100 µg of GnRH + controlled internal drug release device (CIDR) containing 1.38 g of progesterone (P4) on d 1, 25 mg of PGF2α on d 8, and CIDR removal + 100 µg of GnRH on d 11. Grass-seed straw was offered for ad libitum consumption. Soybean meal was individually supplemented at a daily rate of 1 kg/cow (as-fed basis). Moreover, 3WK was supplemented on d 0, 2, 4, 7, 9, 11, 14, 16, and 18 whereas 1WK was supplemented on d 4, 11, and 18. Blood samples were collected from 0 (before) to 72 h after supplementation on d 11 and 18 and analyzed for plasma urea-N (PUN). Samples collected from 0 to 12 h were also analyzed for plasma glucose, insulin, and P4 (d 18 only). Uterine flushing fluid was collected concurrently with blood sampling at 28 h for pH evaluation. Liver biopsies were performed concurrently with blood sampling at 0, 4, and 28 h and analyzed for mRNA expression of carbamoyl phosphate synthetase I (CPS-I; h 28) and CYP2C19 and CYP3A4 (h 0 and 4 on d 18). Plasma urea-N concentrations were greater (P < 0.01) for 1WK vs. 3WK from 20 to 72 h and greater (P < 0.01) for 1WK vs. D from 16 to 48 h and at 72 h after supplementation (treatment × hour interaction, P < 0.01). Moreover, PUN concentrations peaked at 28 h after supplementation for 3WK and 1WK (P < 0.01) and were greater (P < 0.01) at this time for 1WK vs. 3WK and D and for 3WK vs. D. Expression of CPS-I was greater (P < 0.01) for 1WK vs. D and 3WK. Uterine flushing pH tended (P ≤ 0.10) to be greater for 1WK vs. 3WK and D. No treatment effects were detected (P ≥ 0.15) on expression of CYP2C19 and CYP3A4, plasma glucose, and P4 concentrations, whereas plasma insulin concentrations were greater (P ≤ 0.03) in D and 3WK vs. 1WK. Hence, decreasing frequency of protein supplementation did not reduce uterine flushing pH or plasma P4 concentrations, which are known to impact reproduction in beef cows.

Dietary selenium and nutritional plane alter specific aspects of maternal endocrine status during pregnancy and lactation.

Objectives were to examine effects of selenium (Se) supply and maternal nutritional plane during gestation on placental size at term and maternal endocrine profiles throughout gestation and early lactation. Ewe lambs (n = 84) were allocated to treatments that included Se supply of adequate Se (ASe; 11.5 µg/kg BW) or high Se (HSe; 77 µg/kg BW) initiated at breeding and nutritional plane of 60% (RES), 100% (CON), or 140% (EXC) of requirements beginning on day 40 of gestation. At parturition, lambs were removed from their dams, and ewes were transitioned to a common diet that met requirements of lactation. Blood samples were taken from a subset of ewes (n = 42) throughout gestation, during parturition, and throughout lactation to determine hormone concentrations. Cotyledon number was reduced (P = 0.03) in RES and EXC ewes compared with CON ewes. Placental delivery time tended (P = 0.08) to be shorter in HSe ewes than in ASe ewes, whereas placental delivery time was longer (P = 0.02) in RES ewes than in CON and EXC ewes. During gestation, maternal progesterone, estradiol-17ß, and GH were increased (P < 0.05) in RES ewes and decreased (P < 0.05) in EXC ewes compared with CON ewes. In contrast, maternal cortisol, IGF-I, prolactin, triiodothyronine, and thyroxine were decreased in RES ewes and increased in EXC ewes compared with CON ewes during gestation. Selenium supply did not alter maternal hormone profiles during gestation. During parturition and lactation, maternal hormone concentrations were influenced by both Se and maternal nutritional plane. During the parturient process, HSe ewes tended to have greater (P = 0.06) concentrations of estradiol-17ß than ASe ewes. Three hours after parturition a surge of GH was observed in ASe-RES ewes that was muted in HSe-RES ewes and not apparent in other ewes. Growth hormone area under the curve during the parturient process was increased (P < 0.05) in ASe-RES vs HSe-RES ewes. Ewes that were overfed during gestation had reduced (P < 0.05) estradiol-17ß but greater IGF-I, triiodothyronine, and thyroxine (P < 0.05) compared with RES ewes. Even though ewes were transitioned to a common diet after parturition, endocrine status continued to be affected into lactation. Moreover, it appears that gestational diet may partially affect lactational performance through altered endocrine status.

Maternal dietary intake alters organ mass and endocrine and metabolic profiles in pregnant ewe lambs.

To determine the impacts of Se supply and maternal dietary intake on ewe organ mass and endocrine and metabolic changes throughout gestation, pregnant first parity ewes (n=77) were allocated to 6 treatments in a 2×3 factorial array. Factors included Se [adequate Se (ASe; 9.5µg/kg BW) vs. high Se (HSe; 81.8µg/kg BW)] initiated at breeding and dietary intake [60% (RES), 100% (CON), or 140% (EXC) of requirements] initiated on d 50 of gestation. Ewes were individually fed and blood samples from the jugular vein were obtained approximately every 14 d from d 50 until parturition. Maternal Se supply did not impact endocrine or metabolic status. There was a nutritional intake by day interaction for NEFA, blood urea nitrogen (BUN), insulin, triiodothyronine (T3), thyroxine (T4), progesterone (P4), and estradiol-17ß (E2). As expected, with increased maternal intake, NEFA concentrations were reduced. During the last weeks of gestation, BUN and insulin were elevated in EXC compared with RES ewes. Although the pattern of T3 and T4 differed throughout gestation within a treatment group, as maternal intake increased, circulating T3 and T4 were increased. For P4 and E2, as maternal dietary intake increased, there was a reduction in the steroid concentrations in jugular blood. There was only a main effect of maternal nutrition (P<0.001) for cortisol concentrations with EXC ewes having greater concentrations than RES and CON ewes, which did not differ. Although Se is known to influence thyroid hormone metabolism, supranutritional levels during pregnancy did not alter circulating T3 and T4 concentrations. Alterations in maternal endocrine status may have influenced placental transport of nutrients to the developing fetus, which we have shown previously is affected by maternal dietary Se and intake. In addition, the alterations in mammary gland weight that we observed may explain the impact of maternal nutrition on mammary gland function and colostrum production, thereby further impairing growth of developing neonates.

Influence of nitrogen and sulfur intake on bovine uterine pH throughout the luteal phase.

Previous research has reported that diets high in protein and sulfur decreased uterine pH in cattle. The objective of this study was to determine the effect of high N and high S intake on uterine pH. Holstein (n = 15) and Angus-cross (n = 5) heifers (337.5 ± 8.4 kg of BW) were randomly assigned to 1 of 4 diets: control (CON; 13.4% CP and 0.17% S); high nitrogen (HN; CON plus urea supplement); high sulfur (HS; CON plus calcium sulfate); or both high nitrogen and sulfur (HNS). Diets were individually fed at 2.6% of BW on a DM basis using Calan gates and estrus was synchronized to occur on d 13 (d 0 = start of dietary treatment). Blood samples were collected on d -2 and daily (d 1 to 28) at 1400 h to determine concentrations of plasma urea nitrogen (PUN), sulfate (d 1, 4, 8, 12, 16, 20, 24, and 28), and progesterone. Uterine pH was measured on d 16, 20, 24, and 28 (d 3, 7, 11, and 15 of the estrous cycle). There was a treatment, time, and treatment by time interaction (P < 0.01) on concentrations of PUN. There was an effect of treatment (P < 0.01) on concentrations of sulfate, with concentrations being increased in HS compared with CON, HN, and HNS (P < 0.01), and HNS increased compared with CON (P < 0.01) and HN (P < 0.01). Uterine pH was increased in HN and HNS compared with CON (P < 0.02), whereas HS was not different from any treatment (P > 0.11). There was no effect of time (P = 0.26) or treatment by time interaction (P = 0.71) on uterine pH. In summary, uterine pH was increased in HN and HNS compared with CON, whereas HS was intermediate and was associated with increased concentrations of PUN.