Holstein Bull Calves Research Articles

Feeding colostrum or a 1:1 colostrum:milk mixture for 3 days postnatal increases small intestinal development and minimally influences plasma glucagon-like peptide-2 and serum insulin-like growth factor-1 concentrations in Holstein bull calves

This study evaluated how feeding colostrum- or a colostrum-milk mixture for 3 d postnatal affects plasma glucagon-like peptide-2 (GLP-2), serum insulin-like growth factor-1 (IGF-1), and small intestinal histomorphology in calves. Holstein bulls (n = 24) were fed colostrum at 2 h postnatal and randomly assigned to receive either colostrum (COL), whole milk (WM), or a 1:1 COL:WM mixture (MIX) every 12 h from 12 to 72 h. A jugular venous catheter was placed at 1 h postnatal to sample blood frequently for the duration of the experiment. Samples were collected at 1, 2, 3, 6, 9, 11, and 12 h. Following the 12-h meal, blood was collected at half-hour intervals until 16 h and then at 1-h intervals from 16 to 24 h. A 27-h sample was taken, then blood was sampled every 6 h from 30 to 60 h. Again, blood was taken at half-intervals from 60 to 64 h, then at 65 and 66 h, following which, a 2-h sampling interval was used until 72 h. Plasma GLP-2 (all time points) and serum IGF-1 (at time points: 1, 6, 12, 18, 24, 36, 48, and 72 h) were both analyzed. Duodenal, jejunal, and ileal tissues were collected at 75 h of age to assess histomorphology and cellular proliferation. Feeding COL, rather than WM, increased plasma GLP-2 by 60% for 2 h and tended to increase GLP-2 by 49.4% for 4 h after the 60-h meal. Insulin-like growth factor-1 area under the curve (from 12 to 72 h) tended to be 27% greater for COL than WM calves but was otherwise unaffected by treatment. Ileal crypts tended to proliferate more with MIX than WM, whereas ileal crypt proliferation did not differ for COL compared with MIX or WM and was not different between treatments in the proximal jejunum. Villi height was increased 1.8 and 1.5× (COL and MIX vs. WM) in the proximal and distal jejunum, respectively, whereas MIX duodenal and ileal villi height tended to be 1.5 and 1.4× that of WM. Crypt depth did not differ in any region. Surface area of the gastrointestinal tract was reduced for WM by 60 and 58% (proximal jejunum) and 38 and 52% (ileum) relative to COL and MIX and was 54% less than MIX in the distal jejunum. Overall, extended COL feeding minimally increased plasma GLP-2 and serum IGF-1 compared with WM feeding. As COL and MIX similarly promoted small intestinal maturation, feeding calves transition milk to promote intestinal development could be a strategy for producers.

Health and growth of veal calves provided a fatty acid supplement and a dry teat

Veal calves are at a high risk of disease early in life, which can lead to poor growth. Research is needed to determine interventions that can reduce disease and promote the growth of veal calves. The aim of this study was to determine the effects of fatty acid supplementation and the provision of a dry teat on the incidence of bovine respiratory disease (BRD), navel inflammation, and diarrhea, as well as calf growth. Upon arrival to a commercial veal facility (d 0), 240 Holstein bull calves from 2 cohorts were randomly assigned to 4 treatments using a 2 × 2 factorial arrangement (n = 60/treatment): (1) milk replacer (MR) without fatty acid supplementation and without access to a dry teat (control), (2) MR with fatty acid supplementation (NeoTec5g, Provimi, Brookville, OH) and without access to a dry teat (FAS), (3) MR without fatty acid supplementation and with access to a dry teat (TT), or (4) MR with fatty acid supplementation and with access to a dry teat (FAS+TT). Calves were housed in individual pens from 0 to 9 wk and then paired by treatment at wk 9. Milk replacer was fed twice daily using a step-up program. Fatty acid supplement was added to milk replacer at a feeding rate of 0.5 g/kg of body weight per head per day for the FAS and FAS+TT groups. Health exams were conducted twice weekly for 6 wk to diagnose BRD, navel inflammation, and diarrhea. Body weight, body condition score (BCS), and structural measurements were recorded at wk 0, 5, and 10. Average daily gain (ADG) was calculated for wk 1 to 5, wk 5 to 10, and wk 1 to 10. Health data were analyzed using logistic regression and are reported as relative risk. Body weight, BCS, and structural measurements were analyzed using repeated-measures ANOVA, and ADG was analyzed using a generalized linear model. There was no effect of FAS, TT, or their interaction on body weight or BCS. There was no effect of FAS on ADG wk 1 to 5, wk 5 to 10, or wk 1 to 10. There was a tendency for TT to decrease ADG from wk 1 to 5 but not from wk 5 to 10 or wk 1 to 10. There was a tendency for the interaction of FAS and TT to decrease ADG for wk 1 to 5 but not for wk 5 to 10 and wk 1 to 10. There was no effect of FAS, TT, or their interaction on the risk of BRD, diarrhea, or navel inflammation. We saw no effect of our interventions on calf health or growth. More research is needed to determine whether other factors, such as failure of passive transfer, poor ventilation, barren housing, and low milk allowance in the first few weeks after arrival, may have affected the efficacy of our interventions.

Short communication: Upper critical temperature-humidity index for dairy calves based on physiological stress variables

In this study, upper critical values of the temperature-humidity index (THI) were determined in Holstein bull calves (n = 16) based on noninvasive physiological variables. Meteorological and animal-based data were recorded for a 4-d period following a 24-h habituation. The estimated upper critical THI values based on the assessment of respiratory rate, rectal temperature, ear temperature, heart rate, and salivary cortisol concentrations were 82.4, 88.1, 83.0, 78.3, and 88.8, respectively. We inferred that welfare of young calves may be compromised above a THI of 78 and that calves experience significant heat stress above a THI of 88. Based on the present findings, upper critical THI should be considered to minimize the duration of impaired welfare during summer heat stress episodes.

451 Late-Breaking: Saccharomyces cerevisiae boulardii stimulates immunoglobulin A production in the gut of neonate dairy calves

Abstract Calves are born with a naïve immune system at birth and undergo rapid changes in terms of immunity. This study focused on evaluating the effect of S. boulardii CNCMI-1079 (SCB) supplementation from birth on immunoglobulin A (IgA) production in the gut of neonate dairy calves. Holstein bull calves (n = 20) were delivered naturally and immediately removed from the dam, and housed in individual straw-bedded pens with a random assignment to a treated group (supplementation with SCB: 10 × 109 CFU/d in each morning meal) or a control group (no supplementation). All the calves received two first meals of a standardized colostrum at 2 h and 12 h after birth followed by two meals per day of milk replacer (7.5% of birth BW; 260 g/kg CP; 160 g/kg crude fat, at 150g/L). Calves were euthanized at 7 d of age and samples from jejunum, ileum and colon were collected. The concentration of IgA and the gene expressions of polymeric Ig receptor (pIgR) and a receptor of plasma B cells (CD79α) were compared between the two groups and among the locations. The expression of CD79α was up-regulated in the ileum compared to jejunum and colon tissues (P < 0.01), whereas pIgR down-regulated in the ileum compared to jejunum and colon (P < 0.01). The supplementation of SCB increased the IgA concentration in the ileum and colon of treated calves compared to the control calves (respectively 1.98 ± 0.09 vs 1.18 ± 0.15 mg/g DM of ileum content, and 1.45 ± 0.14 vs 0.59 ± 0.06 mg/g DM of colon content, P < 0.001). The results suggest that one of the mechanisms by which SCB exerts an immunoprotective effect in the gut is by stimulation of IgA’s secretion which may play a major role in mucosal protection in neonatal calves.

Different milk replacers alter growth performance and rumen bacterial diversity of dairy bull calves

The diets of calves predominantly comprise milk and the start of solid feed intake triggers a critical process in terms of the activity of the rumen microbiota and ruminal fermentation. However, little is known about alterations of the rumen bacterial diversity in dairy bull calves induced by milk replacers and other starters, such as solid pellets or hay. Thus, we analyzed the effects of partially replacing milk with pellets and Chinese wildrye in the diet on the growth performance and rumen bacterial composition in dairy bull calves. Thirty-two newborn Holstein bull calves were randomly allocated to four dietary treatments: group MFC was fed milk replacer, pellet diet, and Chinese wildrye; group MFR was fed milk replacer and Chinese wildrye; group MCO was fed milk replacer and pellet diet; and group MIL was fed milk replacer only. The diets were provided for 1 week before the trial and for 7 weeks in the experiment. At the end of the experiment, three calves from each treatment were slaughtered to determine the growth performance and slaughter traits, and the rumen bacteria composition was analyzed by 16S rRNA gene sequencing. The body weight, withers height, body length, and heart girth were higher (P < 0.05) in the MCO and MFC groups compared with the MIL and MFR group. In addition, the fresh weights of the liver, spleen, lung, kidney, rumen, reticulum, and abomasum were higher (P < 0.05) in the MCO and MFC groups. The sequencing results showed that at both the phylum and genus levels, the MIL and MFR groups clustered together, whereas the MCO and MFC groups clustered together. Bacteroidetes and Proteobacteria were more abundant in the MIL and MFR groups, whereas Firmicutes were more abundant in the MFC and MCO groups. The abundances of genera such as Dialister, Megasphaera, Acidaminococcus, and Mitsuokella were higher (P < 0.05) in the MCO and MFC groups. Our results showed that the growth performance of calves could be increased by feeding milk replacer with pellet diet, which may have been associated with changes in the diversity or abundances of rumen bacteria, especially bacteria from the phylum Firmicutes.

Effect of antibiotic treatment in preweaned Holstein calves after experimental bacterial challenge with Pasteurella multocida

The primary objective of this randomized controlled challenge study was to investigate the effect of ampicillin on ultrasonographic (US) lung consolidation after experimental challenge with Pasteurella multocida in preweaned dairy calves. The secondary objectives were to determine whether ampicillin affected respiratory score, gross consolidation, or the detection of P. multocida in lung tissue at postmortem exam (PME). Holstein bull calves (n = 39) were transported to the University of Wisconsin-Madison School of Veterinary Medicine isolation facility at the mean (±SD) age of 52 ± 6 d. After a 7-d acclimation period, 30 calves were inoculated intratracheally with 1010 cfu of ampicillin-sensitive P. multocida. Lung US and respiratory scoring were performed 2, 6, 12, and 24 h post-challenge, then US once daily and respiratory scoring twice daily until d 14. Calves were randomized to receive ampicillin [n = 17, treatment (TX), 6.6 mg/kg i.m. once daily for 3 d] or placebo [n = 11, control (CON), saline, equal volume, i.m. once daily for 3 d] when ≥1 cm2 of lung consolidation was observed and ≥6 h had elapsed since challenge. Lung lesions ≥1 cm2 were considered positive for consolidation. Calves were respiratory score positive if ≥2 in 2 or more categories based on the Wisconsin respiratory health score chart. Area under the curve (AUC) was calculated for US score and respiratory score as a proxy for time with consolidation and clinical respiratory disease, respectively. Gross lung lesions and pathogens were quantified following PME. At the time of first treatment, consolidation had developed in 28/30 calves (TX, n = 17; CON, n = 11) and 6% (1 out of 17) of TX and 9% (1 out of 11) of CON calves had a positive respiratory score. The TX calves had a significantly lower median (interquartile range given in parentheses) AUC for US score [TX: 23 (20, 29), CON: 47 (33, 53)], whereas mean AUC for respiratory score was not different between groups (TX: 93 ± 28, CON: 96 ± 17). On d 14, 70% (12 out of 17) of TX and 100% (11 out of 11) of CON calves had lung consolidation, and 24% (4 out of 17) of TX and 27% (3 out of 11) of CON calves had clinical respiratory disease. On PME, median consolidation was 10% (6, 15) for TX and 10% (2, 28) for CON calves. Lung cultures were positive for P. multocida in 77% (13 out of 17) of TX and 91% (10 out of 11) of CON calves. Lung health benefited from a 3-d ampicillin therapy, but benefits were short-lived. Treatment failures might be due to incomplete resolution of the initial lung infection. Future studies are needed to optimize TX strategies to improve long-term lung health.

Effects of recombinant bovine interleukin-8 (rbIL-8) treatment on health, metabolism, and lactation performance in Holstein cattle III: Administration of rbIL-8 induces insulin resistance in bull calves

Our previous work has suggested that recombinant bovine interleukin-8 (rbIL-8) treatment might influence cow metabolism. Therefore, this study was conducted to initially assess the effects of systemic administration of rbIL-8 on response to a glucose challenge, blood metabolites, insulin, growth hormone, insulin-like growth factor-1, immune cell populations, and inflammatory parameters in Holstein bull calves. Calves from 30 ± 6 d of life were individually housed and randomly allocated to 1 of 2 treatment groups: rbIL-8 (rbIL-8, n = 10) and control (CTR, n = 8). Calves assigned to the rbIL-8 group received 1 s.c. injection (d 1, 0900 h) and 6 i.v. injections (d 1 at 1600 h, d 2 and 3 at 0900 h and 1600 h, and d 4 at 0900 h) of rbIL-8 (4 μg/kg of body weight), whereas the CTR group received 2 mL of sterile saline solution at each time point. Day of enrollment was considered as d 1, and the study duration was 10 d. Insulin concentrations and whole-body glucose disappearance were evaluated by an i.v. glucose tolerance test conducted at 12 h and 7 d following the last rbIL-8 injection. Rectal temperature and blood samples were collected on d 1, 2, 3, and 4 at -30 (before treatment, 0830 h), 30, 60, 120, 240, and 360 min relative to treatment, and daily at 0830 h for the rest of the study period. Serum was harvested, and the following parameters were measured: β-hydroxybutyrate (BHB), nonesterified fatty acids, glucose, insulin, plasma urea nitrogen, haptoglobin, and differential blood count. Significant differences were considered when P ≤ 0.05 and a trend if 0.05 <P ≤ 0.10. Serum glucose levels and glucose area under the curve (AUC) did not differ between treatment groups in response to the glucose challenge. However, calves treated with rbIL-8 had greater serum insulin concentration and insulin AUC compared with controls. Administration of rbIL-8 increased rectal temperature (rbIL-8 = 39.3 ± 0.1; CTR = 38.9 ± 0.1°C; ±standard error), BHB concentrations (rbIL-8 = 3.54 ± 0.10; CTR = 2.99 ± 0.12 mg/dL), counts of lymphocytes (rbIL-8 = 4.52 ± 0.12; CTR = 3.84 ± 0.14 × 103/μL), monocytes (rbIL-8 = 0.87 ± 0.03; CTR = 0.67 ± 0.04 × 103/μL), and granulocytes (rbIL-8 = 3.54 ± 0.22; CTR = 2.66 ± 0.24 × 103/μL). We conclude that rbIL-8 induces insulin resistance in Holstein bull calves, accompanied by systemic inflammation and altered blood metabolites and white blood cell populations.

Quantifying transmission of Mycobacterium avium subsp. paratuberculosis among group-housed dairy calves

Johne’s disease (JD) is a chronic enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP), with control primarily aimed at preventing new infections among calves. The aim of the current study was to quantify calf-to-calf transmission of MAP among penmates in an experimental trial. Newborn Holstein bull calves (n = 32) were allocated into pens of 4, with 2 inoculated (IN) calves and 2 calves that were contact exposed (CE). Calves were group-housed for 3 months, with frequent collection of fecal and blood samples and tissue collection after euthanasia. The basic reproduction ratio (R0) was estimated using a final size (FS) model with a susceptible-infected model, based on INF-γ ELISA and tissue culture followed by qPCR. In addition, the transmission rate parameter (β) for new shedding events was estimated using a general linearized method (GLM) model with a susceptible-infected-susceptible model based on culture, followed by qPCR, of fecal samples collected during group housing. The R0 was derived for IN and CE calves separately, due to a difference in susceptibility, as well as differences in duration of shedding events. Based on the FS model, interferon-γ results from blood samples resulted in a R0IG of 0.90 (0.24, 2.59) and tissue culture resulted in a R0T of 1.36 (0.45, 3.94). Based on the GLM model, the R0 for CE calves to begin shedding (R0CE) was 3.24 (1.14, 7.41). We concluded that transmission of MAP infection between penmates occurred and that transmission among calves may be an important cause of persistent MAP infection on dairy farms that is currently uncontrolled for in current JD control programs.

Effects of supplemental butyrate and weaning on rumen fermentation in Holstein calves

The objectives of this study were to determine the effects of the weaning transition and supplemental rumen-protected butyrate on subacute ruminal acidosis, feed intake, and growth parameters. Holstein bull calves (n = 36; age = 10.7 ± 4.1 d; ± standard deviation) were assigned to 1 of 4 treatment groups: 2 preweaning groups, animals fed milk replacer only (PRE-M) and those fed milk replacer, calf starter, and hay (PRE-S); and 2 postweaning groups, animals fed milk replacer, calf starter, and hay without supplemental rumen-protected butyrate (POST-S) or with supplemental rumen-protected butyrate at a rate of 1% wt/wt during the 2-wk weaning transition (POST-B). Milk replacer was provided at 1,200 g/d; starter, water, and hay were provided ad libitum. Weaning took place over 14 d by reducing milk replacer provision to 900 g/d in wk 7, 600 g/d in wk 8, and 0 g/d in wk 9. Rumen pH was measured continuously for 7 d during wk 6 for PRE-S and PRE-M and during wk 9 for POST-S and POST-B. After rumen pH was measured for 7 d, calves were euthanized, and rumen fluid was sampled and analyzed for volatile fatty acid (VFA) profile. Individual feed intake was recorded daily, whereas, weekly, body weights were recorded, and blood samples were collected. Compared with PRE-M, PRE-S calves tended to have a greater total VFA concentration (35.60 ± 11.4 vs. 11.90 ± 11.8 mM) but mean rumen pH was unaffected (6.25 ± 0.22 vs. 6.17 ± 0.21, respectively). Between PRE-S (wk 6) and POST-S (wk 9), calf starter intake increased (250 ± 219 vs. 2,239 ± 219 g/d), total VFA concentrations increased (35.6 ± 11.4 vs. 154.4 ± 11.8 mM), but mean rumen pH was unaffected (6.25 ± 0.22 vs. 6.40 ± 0.22, respectively). Compared with POST-S, POST-B calves had greater starter intake in wk 7, 8, and 9, but POST-B tended to have lower total VFA concentration (131.0 ± 11.8 vs. 154.4 ± 11.8 mM) and lower mean ruminal pH (5.83 ± 0.21 vs. 6.40 ± 0.22). In conclusion, the weaning transition does not appear to affect rumen pH and VFA profile, but supplementing rumen-protected butyrate during the weaning transition increased starter intake and average daily gain. Further, these data suggest that the ability of the rumen to manage rumen pH changes fundamentally postweaning. Why weaned calves with lower rumen pH can achieve higher calf starter intakes is unclear; these data suggest the effect of rumen pH on feed intake differs between calves and cows.

Microbial Inoculum Composition and Pre-weaned Dairy Calf Age Alter the Developing Rumen Microbial Environment.

The objective of this experiment was to determine if dosing pre-weaned calves with enriched ruminal microbiota alters the rumen microbial environment and growth performance. Twenty Holstein bull calves were removed from their dam at birth, fed 3.8 L colostrum within 4 h after birth, and housed individually. Calves were fed pasteurized milk 3×/d from 0 to 7 weeks of age and offered a texturized calf starter ad libitum at 6 days of age. A randomized complete block design with repeated measures and a 2 × 2 factorial arrangement of treatments was used to evaluate responses. Treatments were administered by stomach intubation once per week from 3 to 6 weeks of age and included: 50 mL autoclaved rumen fluid (RF), 50 mL bacterial-enriched RF (BE), 50 mL protozoal-enriched RF (PE); or 50 mL of each BE and PE inoculum. A rumen content composite was collected from 4 rumen fistulated, lactating cows and used to create the inocula. BE inocula were microscopically confirmed to be free of ciliate protozoa before inoculation, while PE contained 2.9 ± 2.2 × 105 protozoa/mL. RF was collected from the calves once per week before 50 mL of the inoculum was administered. Animal performance (e.g., weight gain and dry matter intake) was not altered by inocula type. All calves were microscopically free of rumen ciliates before inoculum administration and calves that did not receive PE remained ciliate-free. Ciliate protozoa were observed in RF from 6, 8, and 6 PE treated calves (n = 10) at weeks 4, 5, and 6, respectively. Ruminal NH3 was lower in PE treated calves (3.3 vs. 6.8 ± 1.0 mM), while ruminal butyrate molar percent was greater in BE treated calves (10.8 vs. 8.3 ± 0.8). Rumen bacterial diversity measures did not differ by treatment at 3–6 weeks. Individual calf bacterial communities from treated calves became temporarily similar to the inocula at 4 weeks of age, but these communities diverged from the inocula at 5 weeks. This study provides new information about two types of rumen-derived inocula and insight into the challenges of directing the rumen microbial environment in the pre-weaned calf.

Effectiveness of zinc supplementation on diarrhea and average daily gain in pre-weaned dairy calves: A double-blind, block-randomized, placebo-controlled clinical trial.

The objective of this clinical trial was to evaluate the effectiveness of zinc supplementation on diarrhea and average daily weight gain (ADG) in pre-weaned dairy calves. A total of 1,482 healthy Holstein heifer and bull calves from a large California dairy were enrolled at 24 to 48 hours of age until hutch exit at approximately 90 days of age. Calves were block-randomized by time to one of three treatments: 1) placebo, 2) zinc methionine (ZM), or 3) zinc sulfate (ZS) administered in milk once daily for 14 days. Serum total protein at enrollment and body weight at birth, treatment end, and hutch exit were measured. Fecal consistency was assessed daily for 28 days post-enrollment. For a random sample of 127 calves, serum zinc concentrations before and after treatment and a fecal antigen ELISA at diarrhea start and resolution for Escherichia coli K99, rotavirus, coronavirus, and Cryptosporidium parvum were performed. Linear regression showed that ZM-treated bull calves had 22 g increased ADG compared to placebo-treated bulls (P = 0.042). ZM-treated heifers had 9 g decreased ADG compared to placebo-treated heifers (P = 0.037), after adjusting for average birth weight. Sex-stratified models showed that high birth weight heifers treated with ZM gained more than placebo-treated heifers of the same birth weight, which suggests a dose-response effect rather than a true sex-specific effect of ZM on ADG. Cox regression showed that ZM and ZS-treated calves had a 14.7% (P = 0.015) and 13.9% (P = 0.022) reduced hazard of diarrhea, respectively, compared to placebo-treated calves. Calves supplemented for at least the first five days of diarrhea with ZM and ZS had a 21.4% (P = 0.027) and 13.0% (P = 0.040) increased hazard of cure from diarrhea, respectively, compared to placebo-treated calves. Logistic regression showed that the odds of microbiological cure at diarrhea resolution for rotavirus, C. parvum, or any single fecal pathogen was not different between treatment groups. Zinc supplementation delayed diarrhea and expedited diarrhea recovery in pre-weaned calves. Additionally, zinc improved weight gain differentially in bulls compared to heifers, indicating a research need for sex-specific dosing.

Form of calf diet and the rumen. II: Impact on volatile fatty acid absorption

Diet is known to affect rumen growth and development. Calves fed an all-liquid diet have smaller and less developed rumens and a decreased ability to absorb volatile fatty acids (VFA) compared to calves fed both liquid and dry feed. However, it is unknown how rumens respond when challenged with a defined concentration of VFA. The objective of this study was to assess the effects of 2 different feeding programs on VFA absorption in preweaned calves. Neonatal Holstein bull calves were individually housed and randomly assigned to 1 of 2 diets. The diets were milk replacer only (MRO; n = 5) or milk replacer with starter (MRS; n = 6). Diets were isoenergetic (3.87 ± 0.06 Mcal of metabolizable energy per day) and isonitrogenous (0.17 ± 0.003 kg/d of apparent digestible protein). Milk replacer was 22% crude protein, 21.5% fat (dry matter basis). The textured calf starter was 21.5% crude protein (dry matter basis). Feed and ad libitum water intakes were recorded daily. Calves were exposed to a defined concentration of VFA buffer (acetate 143 mM, propionate 100 mM, butyrate 40.5 mM) 6 h before euthanasia on d 43 ± 1. Rumen fluid samples were obtained every 15 to 30 min for 6 h to measure the rate of VFA absorption. Rumen tissues were obtained from the ventral sac region and processed for morphological and immunohistochemical analyses of the VFA transporters monocarboxylate transporter 1 (MCT1) and 4 (MCT4). Body growth did not differ between diets, but empty reticulorumens were heavier in MRS than MRO calves (0.67 vs. 0.39 ± 0.04 kg) and MRS calves had larger papillae areas (0.76 vs. 15 ± 0.08 mm2). We observed no differences between diets in terms of the abundance of MCT1 and MCT4 per unit area. These results indicate that the extrapolated increase in total abundance of MCT1 or MCT4 in MRS calves was not due to increased transporter density per unit area. Modeled VFA absorption metrics (flux, mmol/h, or 6 h absorbed VFA in mmol) were not different across diets. These results demonstrate that the form of calfhood diet, whether solely MR or MR and starter, does not alter VFA absorption capacity when the rumen is exposed to a defined concentration of VFA at 6 wk of age.

Form of calf diet and the rumen. I: Impact on growth and development

Preweaning diet is known to affect rumen tissue appearance at the gross level. The objectives of this experiment were to investigate effects of different preweaning diets on the growth and development of the rumen epithelium and on putative rumen epithelial stem and progenitor cell measurements at the gene and cell levels. Neonatal Holstein bull calves (n = 11) were individually housed and randomly assigned to 1 of 2 diets. The diets were milk replacer only (MRO; n = 5) or milk replacer with starter (MRS; n = 6). Diets were isoenergetic (3.87 ± 0.06 Mcal of metabolizable energy per day) and isonitrogenous (0.17 ± 0.003 kg/d of apparent digestible protein). Milk replacer was 22% crude protein, 21.5% fat (dry matter basis). The textured calf starter was 21.5% crude protein (dry matter basis). Water was available ad libitum and feed and water intake were recorded daily. Putative stem and progenitor cells were labeled by administering a thymidine analog (5-bromo-2'-deoxyuridine, BrdU; 5 mg/kg of body weight in sterile saline) for 5 consecutive days and allowed a 25-d washout period. Calves were killed at 43 ± 1 d after a 6 h exposure to a defined concentration of volatile fatty acids. We obtained rumen tissue from the ventral sac and used it for immunohistochemical analyses of BrdU (putative stem and progenitor cells) and Ki67 (cell proliferation), gene expression analysis, and morphological measurements via hematoxylin and eosin staining. Epithelial stem and progenitor cell gene markers of interest, analyzed by real-time quantitative PCR, were β1-integrin, keratin-14, notch-1, tumor protein p63, and leucine-rich repeat-containing G protein-coupled receptor 5. Body growth did not differ by diet, but empty reticulorumens were heavier in MRS calves (MRS: 0.67 ± 0.04 kg; MRO: 0.39 ± 0.04 kg). The percentage of label-retaining BrdU basale cells was higher in MRO calves than in MRS calves (2.0 ± 0.3% vs. 0.3 ± 0.2%, respectively). We observed a higher percentage of basale cells undergoing proliferation in MRS calves than in MRO calves (18.4 ± 2.6% vs. 10.8 ± 2.8%, respectively). Rumen epithelial gene expression was not affected by diet, but the submucosa was thicker in MRO calves and the epithelium and corneum/keratin layers were thicker in MRS calves. Presumptive stem and progenitor cells in the rumen epithelium were identifiable by their ability to retain labeled DNA in the long term, changed proliferative status in response to diet, and likely contributed to observed treatment differences in rumen tissue thickness.

Calves, as a model for juvenile horses, need only one sprint per week to experience increased bone strength.

Previous research has determined that maintaining young animals in stalls is detrimental to their bone health, while the addition of 50 to 82-m sprints 5 d/week aids in counteracting the reduction of bone strength from confinement. The current research aims to determine if 1 or 3 d/week of sprinting affords the same benefits to bone as 5 d/week of sprinting compared to animals confined with no sprinting. Twenty-four Holstein bull calves were obtained from the Michigan State University Dairy Cattle Teaching and Research Center. At 9 wk of age, calves were randomly assigned to treatments of 1, 3, or 5 d/week of sprint exercise, or to the confined control group sprinted 0 d/week. Each treatment had 6 calves. Individual sprinting bouts included a single sprint down a 71-m concrete aisle. For the duration of the 6-wk study, calves were housed at the MSU Beef Cattle Teaching and Research Center in stalls which afforded calves room to stand, lay down, and turn around. Serum was collected weekly via jugular venipuncture to obtain concentrations of osteocalcin (OC) and C-telopeptide crosslaps of type I collagen (CTX-1)—markers of bone formation and degradation, respectively. Sprints were videotaped weekly to determine stride frequency and sprint velocity. On day 42, calves were humanely euthanized at the Michigan State University Meat Lab and both front limbs were immediately harvested. Computed tomography scans and mechanical testing were performed on the left fused third and fourth metacarpal bones. Serum OC concentration was greatest for calves sprinted 5 d/week (P < 0.001). Calves sprinted 5 d/week had both greater stride frequency (P < 0.05) and lower sprint velocity (P < 0.05). All exercise treatments experienced greater dorsal cortical widths compared to control animals (P < 0.01). Through mechanical testing, fracture forces of all sprinting treatments were determined to be greater than the control treatment (P < 0.02). Results from this study support that sprinting 1, 3, or 5 d/week during growth can increase bone health and cause favorable alterations in bone markers. While all exercise treatments had over a 20% increase to fracture force, calves sprinted 1 d/week sprinted only 426 m over the 6-wk study and still experienced over a 20% increase in bone strength compared to confined calves. This study demonstrates the remarkably few strides at speed needed to enhance bone strength and emphasizes the danger to skeletal strength if sprinting opportunities are not afforded.

Short communication: Effect of manipulating fatty acid profile on gluconeogenic gene expression in bovine primary hepatocytes

During the peripartum period, dairy cows experience both an increase in circulating fatty acid (FA) profile and a change in circulating FA profile, which have been shown to alter regulation of gluconeogenic genes. The objective was to quantify gene expression of key enzymes involved in gluconeogenesis and FA transport into the mitochondria in primary hepatocytes in response to exposure to an FA mixture mimicking what is circulating in a transition dairy cow with or without enrichment of C16:0, C18:0, and C18:1. Primary hepatocytes were isolated from 4 Holstein bull calves 3 d of age (± standard deviation 2 d) and cultured. Twenty-four hours after plating, treatments were applied to the cells for 24-h incubation. Treatments consisted of (1) control (1% BSA), (2) 0.75 mM FA cocktail (3% C14:0, 27% C16:0, 23% C18:0, 31% C18:1, 8% C18:2, and 8% C18:3 to mimic the FA profile of dairy cattle at calving), (3) 0.90 mM FA cocktail, (4) 0.75 mM FA cocktail + 0.15 mM C16:0, (5) 0.75 mM FA cocktail + 0.15 mM C18:0, and (6) 0.75 mM FA cocktail + 0.15 mM C18:1. After harvest in Trizol (Life Technologies, Carlsbad, CA), samples were stored at -80°C until RNA extraction, purification, and reverse transcription. Abundance of mRNA was measured using quantitative real-time PCR. Expression of genes of interest [carnitine palmitoyltransferase 1A, pyruvate carboxylase, cytosolic phosphoenolpyruvate carboxykinase (PCK1), mitochondrial phosphoenolpyruvate carboxykinase, and glucose-6-phosphatase] was calculated relative to the average abundance of 2 reference genes (ribosomal protein L32 and glyceraldehyde 3-phosphate dehydrogenase), which were the most stable out of 3 tested. Data were analyzed using PROC MIXED (SAS version 9.4; SAS Institute, Cary, NC) with the fixed effect of treatment and calf in the random statement. The addition of FA compared with the 1% BSA treatment increased the expression of carnitine palmitoyltransferase 1A and cytosolic PCK1. Enrichment with individual FA did not further regulate pyruvate carboxylase or PCK1 beyond that achieved by the basal profile. These results suggest that shifts in circulating FA profile within a biological range, without a difference in the total FA concentration, have minimal effects on transcriptional regulation of hepatic gluconeogenic genes in primary bovine hepatocytes.

Weaning methods affect ruminal methanogenic archaea composition and diversity in Holstein calves

The objective of the present study was to examine the effect of different weaning methods on the ruminal methanogenic archaea composition and diversity in Holstein calves. Thirty-six newborn Holstein bull calves were assigned to 1 of 3 treatments: (1) conventional weaning (d 56) and fed a high proportion of solid feed (CWS); (2) conventional weaning (d 56) and fed a high proportion of liquid feed (CWL); (3) early weaning (d 42) and fed with a high proportion of solid feed (EWS). High-throughput sequencing of the methyl coenzyme M reductase (mcrA) gene, which encodes the α-subunit of methyl coenzyme M reductase — the enzyme that catalyzes the final step in methanogenesis was used to determine the composition and diversity of rumen methanogens. No significant difference (P>0.05) was observed for operational taxonomic units (OTUs) or richness indices, but diversity indices increased (P<0.05) for calves fed high dietary solids. Predominant families across the three treatments were Methanobacteriaceae, Thermoplasmataceae and Methanomassiliicoccaceae. Calves in the EWS treatment had a higher (P<0.05) relative abundance of Methanobrevibacter sp. strain AbM4 and Methanosphaera stadtmanae, while calves in the CWL treatment had a higher (P<0.05) abundance of Methanosphaera sp. strain SM9. A positive (P<0.05) relationship was identified between butyrate and Methanobrevibacter sp. strain AbM4. In conclusion, the composition and diversity of methanogens in the rumen of Holstein calves varied under the different weaning methods. This study identified a positive relationship between butyrate and Methanobrevibacter sp. strain AbM4, potentially reflecting correlations between ruminal fermentation variables and methanogenesis function. These in-depth analyses provide further understanding of weaning methods for intensified production systems.

Enhanced early-life nutrition upregulates cholesterol biosynthetic gene expression and Sertoli cell maturation in testes of pre-pubertal Holstein bulls

Well-fed prepuberal Holstein bulls had larger testes, earlier puberty, higher LH, testosterone and IGF-1, earlier and more proliferating and differentiating Sertoli cells, and greater sperm production potential. The objective was to determine effects of pre-pubertal nutrition on mRNA expression of testicular genes. Holstein bull calves were fed high or low diets (20 or 12% crude protein, respectively and 71.6 or 64.4% Total Digestible Nutrients) from 2 wk, castrated at 8, 16, 24 and 32 wk and testicular mRNA extracted and sequenced. Differential expression of genes mainly occurred at 16 and 24 wk. At 16 wk, functional analysis (DAVID) of DE mRNA revealed common biological processes including “cholesterol” and “fatty acid biosynthesis,” with most genes (including HMGCR, HMGCS1, HSD17) upregulated in high-diet bulls (P < 0.05). Major pathways enriched at 16 wk were “cholesterol biosynthesis”, “steroid metabolism” and “activation of gene expression by Sterol regulatory element binding protein (SREBP)” (P < 0.05). In high-diet bulls, mature Sertoli cell marker Connexin 43, was upregulated at 16 wk and immature Sertoli cell marker (AMH) downregulated at 24 wk. There was an indirect interaction between insulin family receptor and most upregulated cholesterol biosynthesis genes. Pre-pubertal nutrition enhanced testicular cholesterol/steroid biosynthesis and Sertoli cell maturation.

Performance and carcass quality of crossbred beef x Holstein bull and heifer calves in comparison with purebred Holstein bull calves slaughtered at 17 months of age in an organic production system

When crossbreeding with beef breeds in organic dairy herds, the use of non-sexed semen will results in both male and female offspring to be used for beef production. The objective of the present study was to compare growth and carcass characteristics of 15 Danish Holstein bull (HB) calves, 15 beef x Danish Holstein crossbred bull (CB) calves and 15 beef x Danish Holstein crossbred heifer (CH) calves reared under organic settings. As beef bred sire a high-merit bull of the Limousine breed was used. Furthermore, the study should generate knowledge which can support future organic beef production. Forty-five Spring-born calves were purchased at 10 private farms at approximately 20 days of age with an average initial body weight of 52.9, 58.2 and 56.1 (s.e.m. 2.6) kg for HB, CB and CH, respectively. Calves were housed indoors in groups of five until weaning off cows’ milk at 3 months of age. Calves were gradually introduced to a grass-silage based ration from 3 to 4 months of age. From 4 to 7 months calves were kept on mixed grass pasture of ryegrass and white clover. From 7 to 13.5 months calves were housed indoor in groups of 5 and fed a mixed ration with low-moderate energy content. From 13.5 months of age, the established groups of 5 animals grazed in a rotational paddock system until slaughter at 17 months. There was no difference between HB, CB and CH in average daily gain (ADG) from birth to weaning, from weaning until turn-out and during pasture from 4 to 7 months. The ADG indoor from 7 to 13.5 months, and during pasture from 13.5 to 17 months differed between treatment groups. The CB had greater ADG than CH for both periods with HB being intermediate (P < 0.02–0.001). The ADG indoor from 7 to 13.5 months were 1.06, 1.10 and 0.97 (s.e.m. 0.02) kg d − 1, and ADG during pasture from 13.5 months until slaughter were 0.88, 1.11, and 0.64 (s.e.m. 0.07) kg d − 1 for HB, CB, and CH, respectively. Final body weights were heavier for CB, intermediate for HB, and lowest for CH (575, 534, and 480 (s.e.m. 12.1) kg, P < 0.01). Overall, ADG from arrival to slaughter was 21% higher for CB than CH (P < 0.001), and the average ADG of the combined heifer and bull crossbreds ((CB+CH)/2) was not different from that of HB. Crossbreeding improved EUROP conformation markedly in both CH (5.3) and CB (7.0) compared with HB (3.0) (P < 0.001), and carcass weight was 4% higher for crossbreds ((CB+CH)/2) compared with HB. There was no difference in EUROP fatness between HB and CB which was low (1.0–1.2), whereas CH had an acceptable fatness (2.9). Carcass lean:fat color (assessed using a Danish 1–5 scale) was high for the bulls (3.9–4.0) but acceptable for CH (3.3, P < 0.01). It is concluded that improvements in carcass conformation and carcass weight coupled with similar overall growth of the combined heifer and bull crossbreds support the efficiency of beef x Holstein crossbreds of both sexes compared with purebred Holstein bulls within an organic system with summer grazing. Furthermore, it proved difficult to finish intact bulls on pasture to fulfill a premium beef cut market that requires higher fatness and less dark meat color than obtained herein.

Effect of extended colostrum feeding on plasma glucagon-like peptide-1 concentration in newborn calves

Glucagon-like peptide-1 (GLP-1) plays a role in the regulation of glucose homeostasis via the stimulation of insulin secretion. The objective of this study was to evaluate the effect of extended colostrum feeding on plasma concentration of GLP-1. Holstein bull calves (n = 27) were fed pooled colostrum at 7.5% of birth body weight at 2 h after birth and then fed mature milk (M), a 50:50 mixture of pooled colostrum and milk (CM), or pooled colostrum (C; n = 9 for each treatment) at 5% of birth body weight at 12 h after birth and every 12 h thereafter until 72 h after birth. Blood samples were obtained before (1 and 2 h after birth) and after (until 72 h after birth; 42 time points) the first colostrum feeding, and plasma concentrations of glucose, insulin, and GLP-1 were measured. Data were analyzed by ANOVA of JMP 13 (SAS Institute Inc., Cary, NC) with treatment, time, and treatment × time interaction as fixed effects. Treatment × time interaction was observed for plasma insulin and glucose concentrations, which were mainly the result of lower concentrations from 1 to 2 d after birth for C compared with M. Conversely, on d 3 after birth, the difference between treatments was not observed for insulin and glucose. For the entire experimental period, plasma GLP-1 concentration was higher for C (2.25 ng/mL) compared with M (1.41 ng/mL) and tended to be higher compared with CM (1.58 ng/mL). A treatment × time interaction was observed for GLP-1, but unlike glucose and insulin, this was mainly the result of higher concentrations from 54 to 72 h after birth (on d 3 after birth) for C compared with M or CM. Postprandial plasma concentration of glucose was not correlated with that of GLP-1 but was positively correlated with that of insulin for the 4-h period after feeding on d 1 (r = 0.30) and d 3 after birth (r = 0.33). Postprandial plasma concentration of GLP-1 was positively correlated with that of insulin for the 4-h period after feeding on d 3 after birth (r = 0.20). These results indicate that extended colostrum feeding may increase plasma GLP-1 concentrations, especially 3 d after birth, but further study is necessary to determine the effect on plasma insulin and glucose concentrations.

Characterizing grooming behavior patterns and the influence of brush access on the behavior of group-housed dairy calves

Group housing allows for dairy calves to perform social grooming behavior; however, the use of mechanical brushes may influence how calves groom themselves and others. Our objectives were, first, to characterize the bout characteristics of brush use, self-grooming, and allogrooming in calves and, second, to evaluate the effects of access to a rotating brush on grooming behavior. Holstein heifer and bull calves (n = 32) were group-housed (4 calves/group) at 2 wk of life (17 ± 3 d of age; mean ± standard deviation) and followed to wk 7 of life. Pens were assigned to receive either a rotating brush (BR; n = 4 pens) or no brush (CON; n = 4 pens). Behavior was recorded continuously for 12 h for 2 focal calves/pen on 2 d during wk 4, 6, and 7 of life. We performed a bout analysis by fitting a mixture of normal distributions to the log10-transformed frequency distribution of the intervals between recorded periods of behavior. We calculated bout criteria for brush use (125.9 s), allogrooming (125.9 s), and self-grooming (a mixture of 3 normal distributions provided the best fit for these data, providing 2 bout criteria: a shorter (micro) bout criterion of 50.1 s and a longer (macro) criterion of 1,000 s). Brush use was consistent across weeks, and calves used the brush for 20.5 ± 6.1 min/12 h observation period, in 31.1 ± 1.7 bouts (mean ± SE). The frequency and duration of allogrooming bouts did not differ between treatments and across time. The BR calves tended to self-groom more than CON calves (16.3 vs. 14.3 min/12 h; BR vs. CON; SE = 0.68), and time spent self-grooming decreased across weeks. The frequency and duration of self-grooming micro bouts did not differ between treatments, but BR calves had shorter, more frequent self-grooming macro bouts (10.58 vs. 9.46 bouts; BR vs. CON; SE = 0.24). In summary, we determined that bout criteria could be fitted to grooming behaviors, which may be useful when characterizing these behaviors in future work, and that providing access to a rotating brush influenced self-grooming behavior in group-housed calves.