Modelling and Molecular Dynamics Predict the Structure and Interactions of the Glycine Receptor Intracellular Domain.

Glycine receptors (GlyRs) are glycine-gated inhibitory pentameric ligand-gated ion channels composed of α or α + ß subunits. A number of structures of these proteins have been reported, but to date, these have only revealed details of the extracellular and transmembrane domains, with the intracellular domain (ICD) remaining uncharacterised due to its high flexibility. The ICD is a region that can modulate function in addition to being critical for receptor localisation and clustering via proteins such as gephyrin. Here, we use modelling and molecular dynamics (MD) to reveal details of the ICDs of both homomeric and heteromeric GlyR. At their N and C ends, both the α and ß subunit ICDs have short helices, which are major sites of stabilising interactions; there is a large flexible loop between them capable of forming transient secondary structures. The α subunit can affect the ß subunit ICD structure, which is more flexible in a 4α2:1ß than in a 4α1:1ß GlyR. We also explore the effects of gephyrin binding by creating GlyR models bound to the gephyrin E domain; MD simulations suggest these are more stable than the unbound forms, and again there are α subunit-dependent differences, despite the fact the gephyrin binds to the ß subunit. The bound models also suggest that gephyrin causes compaction of the ICD. Overall, the data expand our knowledge of this important receptor protein and in particular clarify features of the underexplored ICD.

Age-related change in flicker thresholds with rod- and cone-enhanced stimuli.

PURPOSE: Rod and cone photoreceptor-specific tests can be time-consuming. A new non-invasive test is described. The test is based on the measurement of flicker modulation thresholds with rod- and cone-enhanced visual stimuli, which requires only minimum adaptation time. Here, we investigated how the rod-and cone-mediated flicker thresholds vary with age. METHODS: Monocular thresholds with rod and cone-enhanced stimuli were measured in 140 healthy adults, (age range: 18-75 years), foveally (0°) and at four parafoveal locations, at an eccentricity of 5° in each of the four quadrants using five, adaptive, interleaved staircases. Temporal frequencies, stimulus sizes, background luminance and spectral composition, were adjusted appropriately to achieve approximately 1 log unit separation in sensitivity between the rod- and cone-enhanced stimuli. Spectrally calibrated, 'neutral density' filters were used to enable adequate control of display luminance for rod enhanced stimuli. RESULTS: The magnitude of central and parafoveal rod thresholds was significantly higher than the central and parafoveal cone thresholds, respectively (p < 0.001) in both the age groups. However, the rate of increase in central rod thresholds (y = 0.45x-12.79; linear regression equation) was not significantly steeper than the rate of increase in central (y = 0.29x-8.53) cone thresholds (p = 0.15). Centrally, cone thresholds showed a better correlation with rod central thresholds for the age > 45 years (Spearman correlation, ρ = 0.74, p < 0.001) compared to age ≤ 45 years (ρ = 0.41, p < 0.001). CONCLUSIONS: Thresholds with rod- and cone-enhanced stimuli are largely invariant below 45 years of age and increase rapidly above this age. This age-wise normative database can be used as an effective functional-marker to assess photoreceptor sensitivities in retinal diseases.

Evaluation of applying statistical process control techniques to daily average feeding behaviors to detect disease in automatically fed group-housed preweaned dairy calves.

Group housing and computerized feeding of preweaned dairy calves are gaining in popularity among dairy producers, yet disease detection remains a challenge for this management system. The aim of this study was to investigate the application of statistical process control charting techniques to daily average feeding behavior to predict and detect illness and to describe the diagnostic test characteristics of using this technique to find a sick calf compared with detection by calf personnel. This prospective cross-sectional study was conducted on 10 farms in Minnesota (n = 4) and Virginia (n = 6) utilizing group housing and computerized feeding from February until October 2014. Calves were enrolled upon entrance to the group pen. Calf personnel recorded morbidity and mortality events. Farms were visited either every week (MN) or every other week (VA) to collect calf enrollment data, computer-derived feeding behavior data, and calf personnel-recorded calf morbidity and mortality. Standardized self-starting cumulative sum (CUSUM) charts were generated for each calf for each daily average feeding behavior, including drinking speed (mL/min), milk consumption (L/d), and visits to the feeder without a milk meal (no.). A testing subset of 352 calves (176 treated, 176 healthy) was first used to find CUSUM chart parameters that provided the highest diagnostic test sensitivity and best signal timing, which were then applied to all calves (n = 1,052). Generalized estimating equations were used to estimate the diagnostic test characteristics of a single negative mean CUSUM chart signal to detect a sick calf for a single feeding behavior. Combinations of feeding behavior signals were also explored. Single signals and combinations of signals that included drinking speed provided the most sensitive and timely signal, finding a sick calf up to an average (±SE) of 3.1 ± 8.8 d before calf personnel. However, there was no clear advantage to using CUSUM charting over calf observation for any one feeding behavior or combination of feeding behaviors when predictive values were considered. The results of this study suggest that, for the feeding behaviors monitored, the use of CUSUM control charts does not provide sufficient sensitivity or predictive values to detect a sick calf in a timely manner compared with calf personnel. This approach to examining daily average feeding behaviors cannot take the place of careful daily observation.

The association between daily average feeding behaviors and morbidity in automatically fed group-housed preweaned dairy calves.

Group housing and computerized feeding of preweaned dairy calves is gaining popularity among dairy producers worldwide, yet disease incidence and detection remain a challenge in these systems. The aim of this prospective observational cohort study was to describe the relationship between morbidity and feeding behavior around the period of illness detection. Calves were enrolled upon entrance to the group pen on 10 farms in Minnesota (n = 4) and Virginia (n = 6) utilizing group housing and computerized feeding from February until October 2014. Morbidity and mortality events were recorded by the calf caregiver. Farms were visited either every week (Minnesota) or every other week (Virginia) to collect calf enrollment data, feeding behavior data, and health records. Daily average feeding behaviors (drinking speed, mL/min; daily consumption, L/d; rewarded visits to the feeder; and unrewarded visits to the feeder) were described both overall and for sick and healthy calf days. Multivariable mixed models were built to assess the differences in daily average feeding behaviors (drinking speed, daily consumption, rewarded visits, unrewarded visits) between matched sick and healthy calves around the time of an illness event (-10 to 10 d). Final models were controlled for calf age, region (Minnesota and Virginia), group size, disease diagnosis, the random effect of farm, and repeated measurements on calf. A stratified analysis was performed by both day from treatment event and disease diagnosis. We enrolled 1,052 calves representing 43,607 calf days over 9 mo. From these, 176 sick calves had a matched control and were carried forward to the matched pair analysis. Fifty-five percent of sick calves (97/176) were treated for diarrhea, 30% (53/176) were treated for pneumonia, and 15% (26/176) were treated for ill thrift. Sick calves drank 183 ± 27 mL/min (mean ± standard error) more slowly, drank 1.2 ± 0.6 L/d less, and had 3.1 ± 0.7 fewer unrewarded visits than control calves on the first day of treatment. These differences began up to 4 d before the calf was detected as sick, and persisted for 7 to 10 d after treatment. However, changes in feeding behaviors varied by disease diagnosed. Rewarded visits were not associated with morbidity status. The results of this study indicate that sick calves change their feeding behavior before and during an illness event, suggesting that feeding behavior may be a useful tool to detect disease onset.

Effect of feeding whole compared with cell-free colostrum on calf immune status: Vaccination response.

Vaccination contributes to improved herd health and production. Boosting immune development at a young age may have long-term effects by enhancing vaccine immune response and efficacy. In the bovine, colostrum is the sole source of maternal immunity, having a substantial effect on health status in the neonate. To date, colostral antibody concentration is used to evaluate colostrum quality. However, colostrum also contains proteins and cells, which may affect immune development and future responses to vaccines. To determine the effect of maternal colostral cells on immune development, 37 female Holstein and Jersey dairy calves were bottle-fed 4 quarts total of whole colostrum (WC) or cell-free colostrum (CFC) at birth. Calves were vaccinated with 2 series of multivalent vaccines. Series A consisted of vaccines given between 1 and 4mo of life. Series B consisted of vaccines given between 5 and 10mo of life. Calf peripheral blood samples were obtained before each vaccination series and monthly for 3mo after each vaccination series. Cellular blood parameters were determined by flow cytometry. Quantitative real-time PCR was used to determine cytokine gene expression in peripheral blood mononuclear cells before vaccination series B and once a month for 2mo after vaccination series B. Calves fed CFC had fewer numbers of B cells in mo 2 after vaccination series A when compared with WC-fed calves. Calves fed CFC had decreased gene expression levels of IL-2 in mo 1 and numbers of CD4(+)CD62L(+)CD45RO(-) and CD4(+)CD62L(+)CD45RO(+) T cells in mo 0 and 1 after vaccination series B as compared with WC-fed calves. Our findings indicate a greater response to vaccines up to 6 to 10mo post-WC feeding when compared with CFC. These data suggest that adoptive transfer of maternal colostral cells at birth has a long-term effect on development of the neonatal immune system.

Growth, intake, and health of Holstein heifer calves fed an enhanced preweaning diet with or without postweaning exogenous estrogen.

Research has shown that changes in nutrition both before and after weaning can affect mammary development. Additionally, estrogen is known to be a potent mammogenic stimulant. Our objectives were to determine effects of altered preweaning feeding and exogenous estradiol postweaning on growth, intake, and health. Thirty-six Holstein heifer calves were reared on (1) a restricted milk replacer (MR) diet fed at 0.44kg powder dry matter (DM)/day [R; 20.9% crude protein (CP), 19.8% fat, DM basis], or (2) an enhanced MR fed at 1.08kg powder DM/d (EH; 28.9% CP, 26.2% fat, DM basis). The MR feeding was reduced 50% during wk 8 to prepare for weaning. Starter was offered after wk 4 but balanced between treatments. Body weight and frame were measured weekly with intakes and health monitored daily. At weaning, a subset of calves were slaughtered (n=6/diet). Enhanced-fed calves had greater carcass, thymus, liver, spleen, and mammary gland (parenchyma and mammary fat pad) weights. The EH calves also had greater average daily gain (ADG) starting during wk 1 (0.36 vs. -0.06kg/d) and lasting through wk 7 (1.00 vs. 0.41kg/d). Remaining calves received estrogen implants or placebo and were slaughtered at the end of wk 10, creating 4 treatments: (1) R, (2) R + estrogen (R-E2), (3) EH, and (4) EH + estrogen (EH-E2). Postweaning ADG was similar between R, EH, and EH-E2 calves, but greater in R-E2 calves than E calves. The EH-E2 calves had the heaviest mammary glands, and R-E2 calves had heavier mammary glands than R calves. The EH calves consumed more MR DM, CP, and fat preweaning. The R-fed calves consumed more starter DM preweaning. Fecal score was greater for EH calves (1.74 vs. 1.50) preweaning, but days medicated did not differ. Fecal scores were lower for R-E2 calves postweaning. Improved preweaning feeding of calves increased body weights and frame measures. Differences in body weights remained postweaning. Enhanced-fed calves showed greater ADG during the preweaning period but not postweaning. Exogenous estrogen may elicit diet-dependent growth responses. Analysis of collected samples will allow determination of cellular and molecular processes responsible for the marked differences in mammary development observed.

Short communication: Comparison of predicted dietary phosphorus balance using bioavailabilities from the NRC (2001) and Virginia Tech model.

The objective of the current work was to use digestion coefficients from the Feng et al. (2015) model (Virginia Tech model) to calculate P bioavailability of common feeds used in dairy production. Compared with the bioavailability calculated by the Virginia Tech model, using the NRC (2001) P absorption coefficient of 0.64 for forages would underestimate the bioavailabilities of alfalfa hay, alfalfa silage, corn silage, grass hay, and mixed mainly legume silage. For concentrates, using the NRC (2001) P absorption coefficient of 0.70 would overestimate the bioavailabilities of corn grain but underestimate bioavailability of high-moisture corn. Two dairy diets were formulated using nutrient values from the NRC (2001): a standard diet that includes minimal by-products, and a by-product diet that has reduced corn and soybean meal which was replaced with corn gluten feed, distillers grains, hominy feed, and wet brewers grains. For each diet, total bioavailable P was calculated using availability values from the NRC (2001) and the Virginia Tech model. Comparison of P balance (the difference between required and bioavailable P) for each diet was made using the 2 sets of bioavailabilities for a reference cow weighing 682 kg, producing 38.6 kg of milk/d (3.5% fat and 3.0% true protein, 100 d in milk), and consuming 23 kg of dry matter/d, yielding an absorbed P requirement of 59.4 g/d. The standard diet supplied 56.69 and 53.52 g of bioavailable P per day using bioavailabilities from the NRC (2001) and Virginia Tech models, respectively, resulting in a P balance of -2.71 and -5.88±0.26 g/d. The by-product diet provided 75.75 and 78.47 g/d of bioavailable P, yielding P balances of 16.35 and 19.07±0.37 g per day, respectively, using the 2 sets of bioavailabilities. Using P bioavailabilities for individual ingredients that were based on the Virginia Tech model resulted in different bioavailable P levels thus resulted in differing dietary P balances in a field setting.

Effect of feeding whole compared with cell-free colostrum on calf immune status: The neonatal period.

Mortality and decreased weight gain resulting from infection and disease in dairy calves are problems within the dairy industry. The bovine neonate relies solely on colostrum to acquire antibodies through passive transfer. To date, colostrum quality is determined by the concentration of antibodies. However, proteins and cells in the colostrum might also enhance immune development in the neonate. To determine the effect of maternal colostral immune cells on calf health and immune status, maternal colostrum was fed either fresh or after lysis of cells by flash-freezing in liquid nitrogen. Thirty-seven female Holstein and Jersey dairy calves were fed 4 quarts total of whole colostrum (WC) or cell-free colostrum (CFC) at birth. Respiratory and fecal scores were measured from birth to d 45 of life. Calf peripheral blood samples were obtained before and after feeding colostrum as well as on d 1, 3, 7, 14, 21, and 28 of life. Peripheral blood mononuclear cells were collected and analyzed for cellular parameters by flow cytometry. Total respiratory scores were greater in CFC-fed calves compared with WC-fed calves on d 38 of life. There were fewer CD4+ T cells and CD4+CD62L+CD45RO- T cells on d 1 and fewer CD4+CD62L+CD45RO+ T cells on d 1 and 3 in CFC-fed calves compared with WC-fed calves. Compared with WC-fed calves, CFC-fed calves had a greater percentage of CD4+CD62L-CD45RO+ T cells on d 0.25, 1, 3, and 7, and a greater percentage of monocytes on d 7. Our data suggest that colostral cells adoptively transfer and enhance neonatal immunity during the first month of life.

The two common polymorphic forms of human NRH-quinone oxidoreductase 2 (NQO2) have different biochemical properties.

There are two common forms of NRH-quinone oxidoreductase 2 (NQO2) in the human population resulting from SNP rs1143684. One has phenylalanine at position 47 (NQO2-F47) and the other leucine (NQO2-L47). Using recombinant proteins, we show that these variants have similar steady state kinetic parameters, although NQO2-L47 has a slightly lower specificity constant. NQO2-L47 is less stable towards proteolytic digestion and thermal denaturation than NQO2-F47. Both forms are inhibited by resveratrol, but NQO2-F47 shows negative cooperativity with this inhibitor. Thus these data demonstrate, for the first time, clear biochemical differences between the variants which help explain previous biomedical and epidemiological findings.

Cow and herd variation in milk urea nitrogen concentrations in lactating dairy cattle.

Milk urea nitrogen (MUN) is correlated with N balance, N intake, and dietary N content, and thus is a good indicator of proper feeding management with respect to protein. It is commonly used to monitor feeding programs to achieve environmental goals; however, genetic diversity also exists among cows. It was hypothesized that phenotypic diversity among cows could bias feed management decisions when monitoring tools do not consider genetic diversity associated with MUN. The objective of the work was to evaluate the effect of cow and herd variation on MUN. Data from 2 previously published research trials and a field trial were subjected to multivariate regression analyses using a mixed model. Analyses of the research trial data showed that MUN concentrations could be predicted equally well from diet composition, milk yield, and milk components regardless of whether dry matter intake was included in the regression model. This indicated that cow and herd variation could be accurately estimated from field trial data when feed intake was not known. Milk urea N was correlated with dietary protein and neutral detergent fiber content, milk yield, milk protein content, and days in milk for both data sets. Cow was a highly significant determinant of MUN regardless of the data set used, and herd trended to significance for the field trial data. When all other variables were held constant, a percentage unit change in dietary protein concentration resulted in a 1.1mg/dL change in MUN. Least squares means estimates of MUN concentrations across herds ranged from a low of 13.6 mg/dL to a high of 17.3 mg/dL. If the observed MUN for the high herd were caused solely by high crude protein feeding, then the herd would have to reduce dietary protein to a concentration of 12.8% of dry matter to achieve a MUN concentration of 12 mg/dL, likely resulting in lost milk production. If the observed phenotypic variation is due to genetic differences among cows, genetic choices could result in herds that exceed target values for MUN when adhering to best management practices, which is consistent with the trend for differences in MUN among herds.

Effects of milk replacer formulation on measures of mammary growth and composition in Holstein heifers.

Overfeeding prepubertal heifers may impair mammary parenchymal growth and reduce milk production, but evidence suggests that increased intake of a high-protein milk replacer before weaning may be beneficial. This study was designed to evaluate effects of milk replacer (MR) composition on mass and composition of mammary parenchyma and fat pad, growth hormone (GH)/insulin-like growth factor-I (IGF-I) axis gene expression, and putative mammary epithelial stem cells. Specifically, we hypothesized that positive effects of faster rates of gain during the preweaning period alter the development, persistence, or activity of populations of putative mammary epithelial stem cells, possibly through involvement of GH/IGF-I axis molecules. Twenty-four newborn heifers were fed 1 of 4 MR diets (n = 6/diet): control [20% crude protein (CP), 21% fat MR fed at 441 g of dry matter (DM)/d], high protein, low fat (28% CP, 20% fat MR fed at 951 g of DM/d), high protein, high fat (27% CP, 28% fat MR fed at 951 g of DM/d), and high protein, high fat+ (27% CP, 28% fat MR fed at 1,431 g of DM/d). Water and starter (20% CP, 1.43% fat) were offered ad libitum. Animals were killed on d 65 and mammary tissue was subjected to biochemical, molecular, and histological examination. No differences in mammary parenchymal mass or composition, with or without adjusting for empty body weight, were detected. Mass was increased and composition of the mammary fat pad was altered by nutrient intake. No diet differences in putative mammary epithelial stem cell abundance or abundance of transcripts for genes of the GH/IGF-I axis were detected. In this study, growth of the mammary epithelium, size of the mammary epithelial stem cell population, and components of the GH/IGF-I axis did not depend on diet. However, an underlying positive correlation between telomerase, a marker of mammary stem cells, and growth of the mammary parenchyma was detected. Implications of diet-induced effects on mammary fat pad and possible effects on subsequent development and function remain to be determined.

Effects of milk replacer composition on growth, body composition, and nutrient excretion in preweaned Holstein heifers.

Twenty-four newborn Holstein heifer calves were fed 1 of 4 milk replacers (MR): control (20% CP, 21% fat; MR fed at 441 g/d); high protein/low fat (HPLF; 28% CP, 20% fat; MR fed at 951 g/d); high protein/high fat (HPHF; 27% CP, 28% fat; MR fed at 951 g/d); and HPHF MR fed at a higher rate (HPHF+; 27% CP, 28% fat; MR fed at 1,431 g/d). Dry calf starter (20% CP, 1.43% fat) composed of ground corn (44.4%), 48% CP soybean meal (44.4%), cottonseed hulls (11.2%), and molasses (1.0%) was offered free choice. Heifers were obtained from a commercial dairy, blocked by groups of 8 in the order acquired, and randomly assigned to treatments within group. Upon arrival at the research farm, heifers were fed the control for 2 feedings. Treatments were imposed when heifers were 4 +/- 1 d of age. Heifers were on study for 61 +/- 1 d. Body weight and body size measures were taken weekly. Four-day total collection of feed refusals, feces, and urine was initiated at 57 +/- 1 d of age. Heifers were slaughtered at the end of the collection period to evaluate body composition. Preplanned contrasts were used to compare control to all, HPLF to HPHF, and HPHF to HPHF+. Heifers fed the control diet consumed more starter than those fed other treatment diets, but their total dry matter intake and apparent dry matter digestibility were lowest. Fecal output was highest in heifers fed the control diet, whereas urine output and urine N excretion were lowest. Nitrogen intake and urine N excretion were greater for heifers fed HPHF+ compared with HPHF but were not affected by MR fat content (HPLF vs. HPHF). Retention (g/d) of N and P was greater in heifers fed all nutrient-dense diets compared with those fed the control diet, but was not improved by increasing fat in the milk replacer (HPLF vs. HPHF) or by increasing the amount fed. Addition of fat to the milk replacer (HPLF vs. HPHF) increased empty body weight fat content without improving average daily gain or frame measures. Increasing the volume fed (HPHF vs. HPHF+) increased growth rate and empty body weight, but HPHF+ heifers were neither taller nor longer and their carcasses contained more fat. Clear improvements in growth and nutrient retention were observed with more nutrient-dense diets, but most of the improvements were seen with the increased protein intake relative to the control MR; adding fat to the high protein MR did not further improve lean tissue gain.

Effects of milk replacer composition on selected blood metabolites and hormones in preweaned Holstein heifers.

We investigated the effects of increasing dietary protein and energy on concentrations of selected blood metabolites and hormones in Holstein heifers. Twenty-four heifers were fed 1 of 4 milk replacer (MR) diets for 9 wk (n = 6/diet): control [20% crude protein (CP), 21% fat MR fed at 441 g of dry matter (DM)/d], HPLF (28% CP, 20% fat MR fed at 951 g of DM/d), HPHF (27% CP, 28% fat MR fed at 951 g of DM/d), and HPHF+ (27% CP, 28% fat MR fed at 1,431 g of DM/d). Heifers were fed twice daily; water and starter (20% CP, 1.43% fat) were offered free choice and starter orts recorded daily. Serum and plasma aliquots from blood samples collected twice weekly after a 12-h fast were analyzed for insulin-like growth factor (IGF)-I, IGF-binding proteins (IGFBP), growth hormone (GH), insulin, glucose, nonesterified fatty acids, triglyceride, and plasma urea nitrogen concentrations. Only plasma glucose, IGFBP-2, and IGFBP-3 were affected by diet. Dietary treatment differences were only noted when the control was compared with the average of the other 3 diets. The addition of fat to the MR (HPLF vs. HPHF) and increased volume of MR (HPHF vs. HPHF+) had no effect on plasma glucose concentration or relative abundance of IGFBP-2 or IGFBP-3. Heifers fed the control diet had less glucose, greater IGFBP-2, and less IGFBP-3 than the average of the other 3 diets. There was a diet by week interaction for IGF-I. Serum IGF-I concentration in control heifers varied in a quadratic manner with a nadir (20 +/- 4 ng/mL) at wk 4, whereas IGF-I increased linearly in heifers on other diets. Both insulin and triglyceride changed over time in a complex pattern (significant linear and quadratic contrast effects). The greatest concentrations were measured at wk 0.5 with nadirs at wk 6 for both insulin and triglyceride. Serum GH concentration decreased in a linear manner from wk 0.5 to wk 9 in all heifers. Relative abundance of IGFBP-2 was quadratic over time with the greatest amount of IGFBP-2 observed at wk 5. With the exception of glucose, IGF-I, IGFBP-2, and IGFBP-3, the blood variables measured were not influenced by treatment. The IGF-I -GH-IGFBP axis requires further study in heifers to deduce effects of nutrition on hypothalamic regulation of metabolism. We expected to see more treatment differences in concentrations of metabolites involved with protein and fat metabolism. It is likely that the diets used in this study were not diverse enough in composition to elicit such changes or that the efficiency of use of absorbed protein and fat was not different in these animals.

Influence of dietary fat and protein on body composition of Jersey bull calves.

Thirty-nine bull calves between 6 and 9 d of age, were assigned to either baseline slaughter or 1 of 4 diets to determine the influence of dietary fat and protein content, at 2 levels of intake, on growth and body composition changes. Calves were assigned to the following diets a 28.5% protein and 16.4% fat milk replacer [MR; 29/16 (n = 9)], 27.3% protein and 33.4% fat MR [27/33 (n = 8)], 20.6% protein and 20.6% fat MR [20/20 (8)], or whole milk [WM (n = 8)]. Calves fed 27/33, 29/16, and WM received 180 g/d of CP to support 650 g of ADG based on predictions from the 2001 NRC. Calves were fed 3 times daily for 4 wk. Weight, hip height, wither height, heart girth, and body length were measured weekly. Weekly plasma samples were analyzed for plasma urea nitrogen, nonesterified fatty acids, and glucose. A subset of calves from each treatment was killed [29/16 (n = 7), 27/33 (n = 6), 20/20 (n = 6), and WM (n = 5)] at the end of wk 4 of treatment; processed for whole-body analysis of fat, protein, ash, and DM; and compared with baseline slaughter calves to estimate composition of empty BW gain. Calves did not differ in average weekly scour score or medication days. Feed efficiency and ADG were greatest for calves fed WM and least for calves fed 20/20; calves fed 29/16 and 27/33 did not differ. Calves fed 27/33 or WM had the greatest % body fat and gained more grams of fat than calves fed other diets. Calves fed 29/16 or 20/20 had similar % fat in empty body as baseline. Differences in % CP, % ash, or % water in empty body and empty BW gain were not detected. Calves fed 27/33 had a trend toward higher NEFA in wk 1 and 2 than calves fed 29/16 or WM. Growth of calves fed 27/33 and 29/16 were similar except that calves fed 29/16 had lower body fat % than calves fed 27/33. Calves on all diets gained less than predicted by the 2001 NRC.

Nitrogen and phosphorus retention and excretion in late-gestation dairy heifers.

The objectives of this study were to evaluate the effects of limit feeding diets containing concentrates or by-products in place of forages on manure and nutrient excretion in growing, gravid heifers. Eighteen Holstein heifers confirmed pregnant were grouped by due date and fed 1 of 3 diets (n = 6 per treatment) for the last 14 wk of pregnancy. Diets were high forage, fed ad libitum (HF); by-product based (BP), fed at the same rate as HF-fed heifers; or low forage (LF), fed at 86% of the HF diet. Diets were designed to supply equal quantities of P, N, and metabolizable energy. Total collection of feces and urine was conducted in wk 14, 10, 6, and 2 prepartum. The HF ration was 90.7% forage, 13.7% crude protein (CP), and contained orchardgrass hay, corn silage, corn grain, soybean meal 44%, and a vitamin-mineral premix. The BP diet was 46.2% forage and 14.0% CP, with 70% of the grain mix space replaced with soybean hulls and cottonseed hulls in a 1:1 ratio, with intake limited to 93% of the dry matter intake (DMI) of HF. The LF ration was 45.3% forage and 17.8% CP, with intake limited to 86% of the DMI of HF. The effect of diet was analyzed with repeated measures, using preplanned contrasts to compare HF with BP and LF with HF and BP. As designed, heifers fed HF and BP had greater DMI than the heifers limit-fed LF, and there was no effect of diet on average daily gain or BW. Intake and digestibility of N were lower, and fecal N excretion was higher, in heifers fed HF and BP than heifers fed LF. Mean feces excretion on both a wet and dry basis was greater for HF heifers compared with BP heifers and less for LF heifers than for HF and BP heifers. Despite differences in urinary output, diet had no effect on urea N excretion, but there was a trend for heifers fed HF and BP rations to excrete less urinary N compared with those fed LF. Compared with HF and BP heifers, LF heifers tended to have lower fecal P excretion and had higher urinary P excretion. Measured manure and urine excretion from heifers fed LF was greater than current American Society of Agricultural and Biological Engineers values, whereas heifers fed HF excreted less manure and urine than predicted. Heifers achieving similar rates of gain from diets differing in forage, grain, and by-product content excreted widely varying quantities of manure.

Influence of pooled colostrum or colostrum replacement on IgG and evaluation of animal plasma in milk replacer.

Newborn Holstein (n = 48) and Jersey (n = 30) calves were studied to compare absorption of immunoglobulin G (IgG) from maternal colostrum (n = 39) or colostrum replacement containing an Ig concentrate derived from bovine serum (n = 39). Calves were also fed milk replacer with (n = 38) or without (n = 40) animal plasma (20% of crude protein) to 29 d of age to determine effect of plasma protein on IgG status, health, and growth. Calves were fed maternal colostrum or colostrum replacement at 1.5 and 13.5 h of age and provided a total of 250 or 249 and 180 or 186 g of IgG for Holsteins and Jerseys fed maternal colostrum or colostrum replacement, respectively. Milk replacer (12.5% DM) was fed at 31% of metabolic birth weight (2 feedings/d). Plasma was sampled at 0 h, 24 h, and weekly to determine IgG by turbidimetric immunoassay. At blood collection, calves were weighed and measured to determine growth. Health scores, fecal scores, and grain intake were measured daily. Plasma IgG at 24 h did not differ between calves fed maternal colostrum (13.78 +/- 0.39 g/ L) and colostrum replacement (13.96 +/- 0.38 g/L). Average daily gain, withers height, hip height, body length, heart girth, health, and incidence of diarrhea were not different between treatment groups. Calves fed maternal colostrum used feed more efficiently than calves fed colostrum replacement. Plasma IgG and performance were not affected by the addition of animal plasma to milk replacer. The colostrum replacement used in this study provided adequate IgG for newborn calves. Animal plasma was an acceptable source of protein but did not enhance growth or immunity under the conditions of this study.

Comparison of uptitration of gliclazide with the addition of rosiglitazone to gliclazide in patients with type 2 diabetes inadequately controlled on half-maximal doses of a sulphonylurea.

Patients with type 2 diabetes who are inadequately controlled on a half-maximal dose of a sulphonylurea may be managed by either increasing the dose of sulphonylurea or adding another agent. In this study, 471 patients inadequately controlled (fasting plasma glucose [FPG] >or=7.0 and

Short communication: effect of pH on absorption of immunoglobulin G in neonatal calves.

We fed newborn calves (n = 60) maternal colostrum (n = 12) or a colostral supplement product derived from edible bovine serum (n = 48). Sodium citrate was added to supplements to achieve a final pH of 7.5, 7.0, 6.0, and 5.0. Calves were fed colostrum (2 L/feeding) or supplement (454 g reconstituted in 2 L of water) at 1.2 and 12.6 h of age, which provided a total of 156 (colostrum) or 90 (supplement) g of IgG. We sampled jugular blood at 0 and 24 h of age to determine plasma IgG. Mean plasma IgG concentrations at 0 and 24 h of age were 0 and 6.7 g/L and were markedly higher in calves fed maternal colostrum compared with supplements (10.7 vs. 6.5 g/L) and were higher in Jersey vs. Holstein calves (8.5 vs. 6.1 g/L). Estimated apparent efficiency of IgG absorption was unaffected by treatment and averaged 20%. These data indicate that pH of colostral supplements between 7.5 and 5.0 do not markedly influence the efficiency of IgG absorption.

Effect of rumen-undegradable protein and energy on growth and feed efficiency of growing Holstein heifers.

Thirty-two Holstein heifers were used to determine the effect of energy and rumen-undegradable protein (RUP) on growth and feed efficiency. Treatment diets were in a 2 x 2 factorial consisting of two energy concentrations (supporting 0.6 or 0.9 kg of average daily gain) and two percentages of RUP (30 or 50% of crude protein). Isonitrogenous diets based on corn silage, alfalfa silage, orchardgrass hay, corn, soybean meal, blood meal, and minerals were fed. Treatment diets were fed until 385 d of age, constituting phase 1 of the trial. During phase 2 (until calving), heifers were housed together and fed a common diet for 0.7 kg of average daily gain. Dry matter intake (kilograms per day) during phase 1 was 4.46, 5.42, 7.38, and 5.95 for heifers fed low energy and low RUP, low energy and high RUP, high energy and low RUP, and high energy and high RUP diets, respectively. Average daily gain (kilograms) during phase 1 was 0.62, 0.74, 1.01, and 0.96, respectively. Dry matter intake and gain were higher for heifers receiving high energy diets. Dry matter efficiency was higher for heifers fed high energy and high RUP diets. Apparent total digestible nutrient efficiency was higher for heifers fed high RUP diets. Average daily gain during phase 2 was 0.53, 0.43, 0.33, and 0.50 kg, respectively. Overall average daily gain was not affected by energy or RUP levels during phase 1 because the slow growth of heifers during phase 1 was compensated by faster growth during phase 2.

Influence of undergradability of protein in the diet on intake, daily gain, feed efficiency, and body composition of Holstein heifers.

Thirty-two Holstein heifers with body weights (BW) between 213 and 231 kg were randomly assigned to one of four treatments for the 50-d trial. Treatments consisted of four percentages of rumen-undegradable protein (RUP) (31, 43, 50, and 55% of total N) at 100% of National Research Council recommendations for total digestible nutrients and crude protein. Total mixed diets composed of corn silage, ground barley straw, soybean meal, blood meal, urea, and minerals were formulated for a mean daily BW gain of 0.60 kg. Ration RUP percentage was varied by shifting protein sources. Mean dry matter intake (grams per kilogram of BW0.75) was 97.6, 84.4, 77.8, and 73.5 for 31% RUP (soybean meal), 43% RUP (blood and soybean meal), 50% RUP (blood meal with urea), and 55% RUP (blood meal) treatments, respectively. Daily gain was 0.84, 0.89, 0.91, and 0.96 kg/d, respectively. Intake of digestible energy (megacalories per kilogram of BW0.75 per day) was 0.28, 0.24, 0.22, and 0.21, respectively, and feed efficiency (megacalories of digestible energy per kilogram of BW gain) was 20.6, 16.1, 15.2, and 13.3, respectively. Dry matter intake (grams per kilogram of BW0.75), digestible energy intake, feed efficiency, daily BW gain, and hip height differed with respect to treatment. There were no differences in growth, wither height, or heart girth because of treatments. Changes in percentage of empty body fat as estimated by urea space procedures was 6.73, 4.67, 6.67, and 7.32, respectively, and did not differ with respect to treatments. These results indicate that increasing the RUP percentage in the diets of growing heifers improves feed efficiency and increases BW gain.