Weaning age influences indicators of rumen function and development in female Holstein calves.

BACKGROUND: Prenatal and postnatal conditions are crucial for the development of calves. Primiparous cows are still maturing during pregnancy, thus competing with the nutritional needs of their offspring. Therefore, mature cows might provide a superior intrauterine condition. Furthermore, weaning calves at an older age might affect them positively as well by reducing stress and offering time for various organs and their functions to develop. We aimed to evaluate effects of mothers' parity and calves' weaning age on gastrointestinal development and corresponding acid-base balance. Fifty-nine female German Holstein calves (about 8 days old) were investigated in a 2 × 2 factorial experiment with factors weaning age (7 vs. 17 weeks) and parity of mother (primiparous vs. multiparous). Calves were randomly assigned to one of these four groups. Animal behavior that was observed included resting, chewing and active behavior. RESULTS: Behavioral patterns were interactively affected by time and weaning age. Rumen sounds per 2 min increased in early-weaned calves during their weaning period. In late-weaned calves a consistently increase in rumen sounds was already recorded before their weaning period. Urinary N-containing compounds (creatinine, hippuric acid, uric acid, urea, allantoin) were interactively affected by time and weaning age. Concentrations of all measured compounds except urea increased during early weaning. All except hippuric acid concentration decreased in early-weaned calves after weaning. In late-weaned calves allantoin and uric acid increased before weaning and did not change during weaning. CONCLUSION: These results suggest that late-weaned calves developed adequate rumen functions and acid-base balance, whereas early-weaned calves might have suffered from ruminal acidosis and catabolism. Weaning calves at 7 weeks of age might be too early for an adequate rumen development.

The involvement of translationally controlled tumor protein during lamb rumen epithelium development.

Early weaning is usually applied to improve the reproductive efficiency of sheep in mutton production, while the development of rumen is of vital importance for sheep weaning age. Translationally controlled tumor protein (TCTP) is a highly conserved protein which participates in multiple tissue and organ development. Thus, we hypothesized that TCTP was involved in sheep rumen development. Histological analyses of sheep rumen epithelium showed that the epithelium formed tough shaped papillae without growing from birth to day 15 of age, after which it rapidly developed to functional epithelia on day 45 of age. We then found TCTP expressed in stratum basale, stratum spinosum and stratum granulosum of rumen epithelium. TCTP protein expression remained at a relative low level from day 0 to day 15 of age, it then significantly increased on day 30 (p < 0.05) and gradually decreased until day 60. Furthermore, to explore the role of TCTP in sheep rumen and its regulation, we found the ratio of Ki67 positive cell in stratum basale cells followed the similar pattern as the expression of TCTP. We also found the ratio of acetate:propionate in rumen fluid decreased from day 30 to day 60 of age (p < 0.05). To conclude, our data indicated that TCTP participated in rumen papillae growth by promoting rumen stratum basale cell proliferation.

Forestomach developmental failure in an 11-month-old Japanese Black steer with severely retarded growth and chronic ruminal tympany.

This study reports findings from the pathological examination of the forestomach of an 11-month-old Japanese Black steer with severely retarded growth (41% of expected weight) and chronic ruminal tympany. The ruminal papillae were weakly formed (0.3-0.5 cm long) and unevenly distributed. The cellulae and cristae reticuli were underdeveloped; the cristae were 0.4-0.7 cm in height and milky white. The keratinized layer in the stratified squamous epithelium was thickened. Ruminal pH was 5.25, and ruminal volatile fatty acid concentration was 11.7 mM. The steer's severely retarded growth was considered to be caused by malnutrition due to developmental and functional failure of the forestomach.

The effect of meloxicam on neonatal dairy calves: Immunoglobulin G uptake and preweaning performance.

Objectives of this study were to determine effects of meloxicam administered in 2 forms on IgG uptake, growth, and health of preweaned calves. Sixteen Holstein bulls and 14 heifers with a body weight (BW) of 44.3 ± 5.24 kg were blocked by birth date in a randomized complete block design. Calves were removed from the dam before suckling, weighed, and randomly assigned to 1 of 3 treatments: (1) colostrum replacer (CR) at 0 h with no meloxicam (control; CON), (2) 1 mg/kg of BW of meloxicam in pill form before CR (PL), or (3) 1 mg/kg of BW of meloxicam mixed in solution with CR (SL). Calves were fed 675 g of dry matter of CR, providing a volume of 3 L and 180 g of IgG. Blood samples were collected at 0 h to analyze initial IgG and ketone concentrations, and at 6, 12, 18, and 24 h to analyze IgG uptake. At 24 h, calves were fed 432 g of dry matter of 24% crude protein milk replacer (MR) split in 2 feedings, and free choice starter and water until 42 d. Weekly blood samples were analyzed for glucose, plasma urea nitrogen, and ketone concentrations. Time of consumption of MR, BW, length, hip and withers height, and heart girth were recorded weekly. All calves achieved adequate transfer of immunity. Meloxicam did not affect apparent efficiency of absorption, serum total protein, or IgG uptake at 6, 18, and 24 h; however, meloxicam-treated calves had lesser IgG concentrations at 12 h (24.40 and 22.59 g/L for PL and SL, respectively) compared with CON (28.47 g/L). Meloxicam treatment did not affect BW. Calves that received PL tended to gain length at a faster rate (0.24 cm/d) than those that received SL (0.19 cm/d). Meloxicam treatment did not affect MR intake, time of consumption of MR, total dry matter intake, or feed efficiency. Meloxicam-treated calves tended to consume more starter (560.4 and 515.4 g/d for PL and SL, respectively) than those that received CON (452.6 g/d). Ketone levels tended to be greater in meloxicam-treated calves (0.15 and 0.17 mmol/L for PL and SL, respectively), suggesting improved rumen development compared with those that received CON (0.12 mmol/L). Meloxicam treatment did not affect plasma urea nitrogen . Glucose concentrations of calves that received PL (73.2 mg/dL) were less than those that received SL (83.3 mg/dL). Results of this study suggest that meloxicam given at 0 h offers positive effects on starter intake, and possibly rumen development, of preweaned dairy calves. Treatment PL, as compared with SL, offered positive results for rumen development, indicated by lower blood glucose levels.

Whole-flint corn grain or tropical grass hay free choice in the diet of dairy calves.

Tropical grass hay feeding is related to improved ruminal health; however, it may decrease energy intake. On the other hand, whole-flint corn grain may be an alternative fiber source in the diet of dairy calves. Forty-two Holstein calves were used in a randomized block design, considering sex, birth date, and weight at 21 d of age, when the supply of whole-flint corn grain or tropical grass hay started. Three component-fed solid diets were compared: (1) starter concentrate only during the preweaning and starter concentrate with free choice of chopped Tifton-85 hay postweaning (SC), (2) starter concentrate with free choice of chopped Tifton-85 hay pre- and postweaning (SCH), and (3) starter concentrate with free choice of whole-flint corn grain pre- and postweaning (SCW). The animals were evaluated from 21 to 84 d of age. Calves were managed equally during the first 21 d, fed with 6 L/d of whole milk and a commercial starter concentrate (46% nonfiber carbohydrates, small particles, and pelleted) ad libitum. After that, milk feeding was reduced to 4 L/d until gradual weaning at 56 d of age. At 56 d of age, 4 animals per treatment were randomly chosen to be slaughtered for digestive tract weight evaluation and to collect tissue for histological analysis of the ruminal wall, duodenum, and cecum, whereas the other 30 animals were weaned and evaluated for a further 22 d when the SC diet also received hay ad libitum. Feed intake was measured daily. Weight gain and metabolic indicators of intermediate metabolism were evaluated weekly. Ruminal fluid was collected at wk 6, 8, 10, and 12 of age. The SCH diet increased the total and starter dry matter intake, and consequently, the average daily gain and body weight at 56 d of age. The SCW diet promoted an increase in propionate and decreased acetate-to-propionate ratio. Morphometric variables were affected by the SCH diet. The postweaning performance was unaffected by solid diets; however, the SCW diet decreased ruminal and fecal pH. Feeding hay, starting at 21 d of age, can stimulate early solid diet intake, promoting better performance and ruminal and intestinal development, when a highly fermentable and small particle pelleted starter is fed.

Effects of 2 colostrum and subsequent milk replacer feeding intensities on methane production, rumen development, and performance in young calves.

A growing need exists for the development of practical feeding strategies to mitigate methane (CH4) emissions from cattle. Therefore, the objective of this study was to evaluate the influence of milk replacer feeding intensity (MFI) in calves on CH4 emission, rumen development, and performance. Twenty-eight female newborn Holstein calves were randomly assigned to 2 feeding groups, offered daily either 10% of the body weight (BW) in colostrum and subsequently 10% of the BW in milk replacer (MR; 10%-MR), or 12% of the BW in colostrum followed by 20% of the BW in MR (20%-MR). In wk 3, half of each feeding group was equipped with a permanent rumen cannula. Both groups were weaned at the end of wk 12. Hay and calf starter (mixture of pelleted grains) were offered from d 1 until wk 14 and 16, respectively. A total mixed ration was offered from wk 11 onward. Feed intake was measured daily and BW, anatomical measures, and rumen size weekly. Methane production and gastrointestinal passage rate were measured pre-weaning in wk 6 and 9 and post-weaning in wk 14 and 22, with additional estimation of organic matter digestibility. Rumen fluid, collected in wk 1, 2, 3, 6, 9, 14, 18, and 22, was analyzed for volatile fatty acid concentrations. Although the experimental period ended in wk 23, rumen volume of 17 calves was determined after slaughter in wk 34. Data was analyzed using ANOVA for the effects of feeding group, cannulation, and time, if applicable. Dry matter intake (DMI) of solid feed (SF) in 20%-MR animals was lower pre-weaning in wk 6 to 10 but mostly higher post-weaning. From wk 6 onward, anatomical measures and BW were greater in 20%-MR animals, and only the differences in body condition score gradually ceased post-weaning. Following the amount of SF intake, 10%-MR calves emitted more CH4 pre-weaning in wk 9, whereas post-weaning the 20%-MR group tended to have higher levels. Methane emission intensity (CH4/BW) was lower pre-weaning in 20%-MR animals but was comparable to the 10%-MR group post-weaning. Methane yield (CH4/DMI of SF) and estimated post-weaning organic matter digestibility were not affected by MFI. Rumen size normalized to heart girth was greater in 10%-MR calves from wk 5 to 10, but differences did not persist thereafter. In wk 34, rumen volume was higher in 20%-MR calves, but normalization to BW revealed no difference between feeding groups. In conclusion, high MFI reduces CH4 emission from calves pre-weaning, although this effect ceases post-weaning.

Influences of starter NDF level on growth performance and rumen development in lambs fed isocaloric and isonitrogenous diets.

Neutral detergent fiber (NDF), as the main component of structural carbohydrate in forage, is a vital factor impacting the performance of young ruminants. The objective of this study was to evaluate the effect of starter NDF level on the rumen fermentation and morphometrics of lambs fed isocaloric and isonitrogenous diets. One hundred Hu sheep lambs (18 days of age and 6.1 ± 0.1 kg body weight [BW]) were randomly assigned to one of the four starter diets differing in their NDF levels on a dry matter (DM) basis: 1) diet with 14% NDF (NDF14), 2) diet with 18% NDF (NDF18), 3) diet with 22% NDF (NDF22), and 4) diet with 26% NDF (NDF26). Lambs were fed milk replacer with designed starters from 21 to 60 d of age and then transitioned to the same starter feed until 90 d of age. Six lambs from each treatment (close to the average BW of the corresponding treatment) were selected and slaughtered at 90 d of age for measuring rumen fermentation, anatomical development, and morphometric characteristics. Lambs receiving NDF22 and NDF26 starter diets had greater (P < 0.05) intake of starter and total DM, and average daily gain during the overall period, thereby heavier (P < 0.05) final weights at 90 d of age compared with those fed NDF14 starter diet. At 90 d of age, lambs fed NDF22 and NDF26 starters had higher rumen pH, followed by lower propionate, higher acetate concentrations (P < 0.05), and a higher acetate-to-propionate ratio compared with those fed NDF14 and NDF18 starters (P < 0.05). No significant differences of the full and empty weight of reticulorumen were observed among treatments; however, the reticulorumen weight expressed as a percentage of the complex stomach weight was lower (P < 0.05) in lambs fed NDF22 and NDF26 starters compared with those fed NDF14 starter. Furthermore, NDF14 lambs had the thickest keratin layer and epithelium compared with those in the other treatments but showed the thinnest muscle layer (P < 0.05). These results suggest that increasing starter NDF levels can ameliorate the rumen fermentation environment and alleviate hyperkeratosis and plaque formation in the rumen epithelium. In summary, including 22% and 26% NDF in the pelleted starter can improve the performance of lambs.

Effects of branched-chain volatile fatty acids and fibrolytic enzyme on rumen development in pre- and post-weaned Holstein dairy calves.

The study evaluated the effects of branched-chain volatile fatty acids (BCVFA) and fibrolytic enzyme (FE) on rumen development in calves. Forty Holstein male calves at the same ages (15 ± 2.5 days of age) and weights (45 ± 3.3 kg of body weight [BW]) were assigned randomly to four groups with a 2 × 2 factorial arrangement of treatments. Supplemental BCVFA (0 g/d or 18 g/d) and FE (0 g/d or 1.83 g/d) were fed to calves. Data were analyzed as a 2 × 2 factorial arrangement random design by the mixed procedure of SAS. The BCVFA × FE interaction was observed for ruminal propionate, blood growth hormone (GH) and insulin-like growth factor-1 (IGF-1), and GH receptor (GHR) and IGF-1 receptor (IGF-1R) expression in the rumen mucosa. Dry matter intake was higher for BCVFA addition. The higher average daily gain and ruminal volatile fatty acids were observed for BCVFA or FE addition. Stomach weight and the length and width of rumen papillae were higher for BCVFA addition. The higher expression of GHR, IGF-1R and 3-hydroxy-3-methylglutaryl-CoA synthase 1 in rumen mucosa, and blood GH and IGF-1 were observed with BCVFA or FE addition. Blood ß-hydroxybutyrate and acetoacetate were higher for BCVFA addition. The results indicated that rumen development was promoted by BCVFA, but was not affected with FE addition in calves.

Impact of early weaning on small intestine, metabolic, immune and endocrine system development, growth and body composition in artificially reared lambs.

AbstractThis study evaluated the effect of early weaning (EW) of artificially reared lambs using a restricted milk replacer (MR) feeding and step-down weaning system on the short- and long-term effects on growth, feed intake, selected blood metabolites and hormones, body composition, and small intestine development. Mixed-sex twin-born 2 to 5 d old lambs were randomly allocated to individual pens and fed MR at 20% of initial individual BW in week 1 and 15% in week 2 followed by weaning off MR by the end of week 4 (EW; n = 16) or week 6 (Control; Ctrl, n = 16) using a step-down procedure. Concentrate starter and fiber diets were offered ad libitum to week 9, then gradually removed over a 10-d period. All lambs were managed as a single group on pasture from weeks 6 to 16 of the trial. Feed intake was recorded daily in the first 6 wk, and BWs recorded weekly. At weeks 2, 4, 6, and 8, and pre- and postclostridial vaccination at week 8, blood samples were collected for analysis of selected blood metabolites, IGF-1, and immune function. Body composition was evaluated in eight animals per group at weeks 4 and 16 after euthanasia, and duodenal samples collected for histomorphometric evaluation. Early weaned lambs had lower DM, ME, CP, and NDF intake than Ctrl lambs at 21, 15, 21, and 36 d of rearing, respectively (P < 0.001), driven by lower intakes of MR from day 15 (P < 0.001) as per the experimental design, and lower total DMI of fiber (P = 0.001) from 21 to 42 d of rearing. Lamb BW tended (P = 0.097) to be lower in EW than Ctrl lambs from 5 to 10 wk of rearing, with lower ADG in EW lambs from weeks 3 to 6 (P = 0.041). Early weaning had negligible effects on duodenal morphology, organ, and carcass weights at weeks 4 and 16. Plasma metabolites (urea nitrogen, triglycerides, NEFA, glucose, and total protein) were similar between groups, while ß-hydroxybutyrate was greater in EW than Ctrl lambs at weeks 4 and 6 (P = 0.018) but not week 8 indicative of early rumen development. Serum IGF-1 tended to be lower in EW than Ctrl lambs from weeks 2 to 6 only (P = 0.065). All lambs developed antibody responses postvaccination and there was no effect of treatment (P = 0.528). The results of this study illustrate that artificially reared lambs can be weaned off MR by 4 or 6 wk of rearing without compromising growth, small intestine morphology, major organ development, and body composition, nor immune function at either 4 (preweaning) or 16 (postweaning) wk of age.

Growth of rumen papillae in weaned calves is associated with lower expression of insulin-like growth factor-binding proteins 2, 3, and 6.

This study aimed to characterize the relationship between the growth of rumen papillae in calves and the mRNA expression of insulin-like growth factor-binding proteins (IGFBPs) in the rumen papillae. The length of rumen papillae, the mRNA expression of IGFBPs in rumen papillae by quantitative real-time PCR, and the presence of insulin-like growth factors I and II (IGF-I and II) by immunohistochemistry (IHC) were analyzed in nine Holstein calves divided into three groups: suckling (2 weeks, n = 3), milk-continued (8 weeks, n = 3), and weaned (8 weeks, n = 3). The length of rumen papillae was greater (p < 0.01) in weaned calves than in suckling and milk-continued calves, whereas the expressions of IGFBP2, IGFBP3, and IGFBP6 genes were lower (p < 0.05) in the rumen papillae of weaned calves than in milk-continued calves. Thus, rumen papillae length and IGFBP2, 3, and 6 expressions were negatively correlated. The IHC analysis showed that IGF-I and IGF-II were present in the rumen epithelium of calves. These results suggested that the growth of rumen papillae after weaning is associated with the induction of IGFs by the low levels of IGFBP2, IGFBP3, and IGFBP6.

Age-associated changes of the intrinsic nervous system in relation with interstitial cells in the pre-weaning goat rumen.

In this study, we investigated the neural changes and their relationships with interstitial cells (ICs) in the rumen of pre-weaning goats by transmission electron microscopy, western blot and immunofluorescence (antibody: general neuronal marker-Protein Gene Product (PGP9.5)/ IC marker-vimentin). The immunofluorescence results showed that PGP9.5-positive reaction was widely distributed in neuronal soma (NS) and nerve fibre (NF). The NSs were observed in the ganglia of the myenteric plexus (MP) but not in the submucosal plexus. The mean optical density (MOD) of the whole of PGP9.5-positive nerves and the protein expression level of PGP.5 in the rumen wall both decreased significantly with age. However an obvious increase MOD of PGP.5-positive NFs within the rumen epithelium were observed. In the MP, the nerves and ICs were interwoven to form two complex networks that gradually tightened with age. Furthermore, NSs and nerve trunks were surrounded by a ring-boundary layer consisting of several ICs that became physically closer with aging. Moreover, ICs were located nearby NFs within the ML, forming connections between ICs, smooth muscle cells and axons. This study describes the pattern of neural distribution and its association with ICs in the developing rumen which shed light on the postpartum development of ruminants.

Effects of ad libitum milk replacer feeding and butyrate supplementation on the epithelial growth and development of the gastrointestinal tract in Holstein calves.

Intensive milk feeding and butyrate supplementation in calves stimulate body growth and affect gastrointestinal development. The aim of the present study was to investigate the synergistic effects of ad libitum milk replacer (MR) feeding and butyrate supplementation of MR on rumen and small intestinal growth and on gene expression in the small intestine related to growth and energy metabolism at weaning. Male Holstein calves (n = 32) received colostrum from birth to d 3 of age and MR either ad libitum (Adl) or restrictively (Res; 6 L of MR/d; 12.5% solids) with (AdlB+, ResB+) or without (AdlB-, ResB-) 0.24% butyrate from d 4 until wk 8 of age. From wk 9 to 10, all calves were weaned and were fed 2 L/d until the end of the trial. Concentrate, hay, and water were freely available. At d 80, calves were slaughtered, volatile fatty acids were measured in rumen fluid, and rumen and small intestine samples were taken for histomorphometric measurements. The expression of mRNA associated with the local insulin-like growth factor (IGF) system and glucose metabolism as well as lactase and maltase activities were measured in the intestinal mucosa. The small intestine was 3 m longer in Adl than in Res. In the atrium ruminis, papilla width was greater in Res than in Adl. Villus circumference, cut surface, and height in the duodenum, proximal jejunum, and ileum were greater in Adl than in Res and in the proximal, mid, and distal jejunum and ileum were greater in calves treated with butyrate. Crypt depth in the duodenum and proximal jejunum was greater in Adl than in Res and in the ileum was smaller in calves treated with butyrate. The villus height:crypt depth ratio was greatest in AdlB+ calves. In the proximal and mid jejunum, IGF1 mRNA abundance was lower in calves treated with butyrate. In the proximal jejunum, INSR mRNA abundance was greater in Res than in Adl. The abundance of PCK2 mRNA was greater in Res than in Adl in the duodenum and was greatest in ResB- in the mid jejunum. Lactase activity tended to be greater in Res than in Adl and after butyrate treatment in the proximal jejunum. The results indicated an elevated growth of the small intestinal mucosa at weaning due to intensive milk feeding and butyrate supplementation, and the local IGF system was involved in intestinal growth regulation. Rumen development was not affected by butyrate supplementation of MR and was slightly delayed due to ad libitum MR feeding.

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 ß1-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.

Effect of selected feed additives to improve growth and health of dairy calves.

The aim was to evaluate the effect of different feed additives on intake, performance, and fecal consistency index (FCI) of dairy calves from 6-60 d of age and its residual effect 15 d after weaning. Fifty Holstein calves (38 ± 1.0 kg BW) were fed 5 L/d of milk plus starter feed until weaning, and corn silage and concentrate after weaning. The treatments were: control (CON), monensin (MON; 30 mg/kg of starter), probiotic E. faecium (PROB; 70 mg/kg of starter), essential oils (EO; 300 mg/kg of starter), or PROB + EO (EOPROB). Fecal score and dry matter intake (DMI) were measured daily, and animals were weighed every 15 d. A DNA extraction from feces was performed to identify the presence of microorganisms (E. coli, Hafnia, Shiguella, Lactobacillus spp, Enterococcus spp, and Enterococcus faecium NCIMB 10415) by PCR. Two 72-h digestibility trials were performed at days 20-28 and 50-56, by total fecal collection. The DMI before weaning was greater for EO (903.0 g/d) compared with MON (794.3 g/d) and EOPROB (783.1 g/d). The FCI decreased during pre-weaning for EO and MON. Average daily gain (ADG) and feed efficiency (FE) did not differ among treatments before weaning. After weaning, DMI and FCI did not differ among treatments. The EO had greater ADG (917.5 g/d) compared with CON (615.8 g/d) and PROB (592.6 g/d). The FE improved with EO (0.72 g/g) over CON (0.36 g/g), MON (0.49 g/g), and PROB (0.36 g/g). The PCR results showed absence of E. faecium NCIMB 10415 in animals fed PROB and CON. Animals fed PROB had greater intake of CP and NDF than animals fed EOPROB. The EO can be added to the dairy calf ration to improve fecal score and increase DMI. The pre-weaning FCI decrease with MON and increase with PROB.

Impact of weaning age on rumen development in artificially reared lambs1.

This study examined the impact of weaning age (4 vs. 6 wk) on rumen morphological and metabolic development in artificially reared lambs. Thirty-two mixed-sex lambs (2 to 5 d old) were randomly allocated to 1 of 2 weaning groups: early weaning (EW; 4 wk) and control (Ctrl; 6 wk). Lambs were individually penned and fed milk replacer (MR; 24% CP and 25% fat, DM basis) at 20% of their corresponding initial BW. Weaning was achieved by gradual reduction of MR allowance over a period of 3 wk using a step-down procedure. Concentrate and meadow hay were offered ad libitum from 1 d of the study until 6 wk, when lambs were transferred to a mixed sward pasture. At week 4, individual DMI were recorded and blood samples collected to measure ß-hydroxybutyrate (BHBA) and NEFA. Eight animals per group were euthanized at week 4 and 16 to evaluate short chain fatty acids (SCFA) and histomorphometry of the rumen walls: dorsal (DS), ventral (VS), dorsal blind (DBS), and ventral blind (VBS) sacs. Linear mixed models were used to analyze the effect of weaning treatments at week 4 and 16, and to compare between timepoints and between rumen sites. Dry matter intake and rumen SCFA profiles were similar between groups (P > 0.10) at week 4 and 16. Plasma concentrations of BHBA were greater (P = 0.03) in EW than in Ctrl lambs, while NEFA did not differ (P > 0.10) between groups at week 4. No effect of weaning age on rumen empty weight, and papillae density, length, width, surface area ratio, and muscle layer thickness at any of the 4 rumen sites was found (P > 0.10) at either week 4 or week 16, except for greater (P = 0.02) papillae epithelium thickness in the DBS at week 4 in EW than Ctrl lambs. Papillae morphology and muscular thickness differed across rumen sites at week 4 and week 16 (P < 0.05), except for papillae density and surface area ratio at 4 week (P > 0.10). Rumen papillae length, width, and muscle layer thickness increased while papillae density and surface area ratio decreased (P < 0.05) between 4 and 16 wk. The results of this study indicate that morphological and physiological development of the rumen can be accelerated to support weaning of artificially reared lambs at 4 wk, using a step-down weaning system. Morphological differences between rumen sacs denote that future studies in lambs evaluating the impact of different diets should involve representative sampling across the rumen rather than a single site to more accurately study rumen development and ontogenic changes.

Effect of weaning age on growth, mammary gland development, and immune function in Holstein Friesian calves fed conserved alfalfa (FiberStart).

This study aimed to evaluate intake, body growth, and the development of the rumen, mammary gland, and immune system in Holstein Friesian calves reared for 100 d on the commercially available feed FiberStart (conserved alfalfa, Medicago sativa; Fiber Fresh Feeds Ltd., Reporoa, New Zealand) and fed calf milk replacer (CMR) for either 56 or 91 d. Eighty calves (40 bulls and 40 heifer calves) were reared indoors in groups (n = 5 of the same sex/pen). All calves were fed 4 L of CMR/d (175 g/L of CMR) in 2 feeds/d for the first 10 d and then 1 feed/d until d 49 or 84. The calves were gradually weaned by d 56 (earlier weaned; n = 8 pens) and d 91 (later weaned; n = 8 pens). All calves were fed FiberStart ad libitum as the only solid feed source from d 1 to 100 of the study. Irrespective of treatment, all calves had similar body weights at d 0 (40.9 ± 3.0 kg) and d 49 (74.2 ± 5.1 kg) of the study. Calf sex had no effect on intake, growth, blood, and immune system parameters. Earlier-weaned calves consumed 18% more solid feed dry matter but had 16% lower body weight gain (28.9 vs. 38.5 kg, respectively) from d 56 to 100 relative to later-weaned calves, resulting in different body weight at 100 d (104 vs. 121 ± 1.3 kg). Although earlier-weaned calves could compensate for the loss of CMR dry matter and crude protein intake from d 56 to 100 by increasing forage intake, they were unable to compensate for the loss of energy from the CMR by increasing solid feed consumption. Plasma ß-hydroxybutyrate concentrations were 52% greater in earlier-weaned calves than in later-weaned calves at d 84, indicating greater metabolic activity of the rumen wall. The duration of CMR feeding had no influence on humoral or cell-mediated immune functions of the calves, as evidenced by a lack of effect on antivaccine antibody responses as well as on immune gene expression. Earlier- versus later-weaned heifer calves had 5% lower mammary gland mass, indicating that greater energy supply increased mammary mass. The results of this experiment demonstrate the ability to artificially rear dairy calves on a conserved alfalfa as the only solid feed. Furthermore, earlier weaning off CMR promotes solid feed intake and an associated increase in blood ß-hydroxybutyrate, an indicator of rumen development, but increasing the duration of CMR feeding improves growth and mammary gland mass by d 100. The implications of these findings on lifetime growth, health, and milk production in dairy heifers warrant further investigation.

Effect of heat-shock protein B7 on oxidative stress in adipocytes from preruminant calves.

Dairy cows with ketosis display excessive lipolysis in adipose tissue. Heat-shock protein B7 (HSPB7), a small heat-shock protein, plays important roles in mediating cytoprotective responses to oxidative stress in rodent adipose tissue. Accordingly, it is assumed that HSPB7 may also play important roles in the antioxidant response in adipose tissue of ketotic cows. Therefore, the aim of this study is to investigate (1) the redox state of adipose tissue in ketotic cows and (2) the role and mechanism of HSPB7 on the regulation of oxidative stress in adipocytes from preruminant calves. An in vivo study consisting of 15 healthy and 15 clinically ketotic cows was performed to harvest subcutaneous adipose tissue and blood samples. In addition, adipocytes isolated from calves were treated with different concentrations of H2O2 (0, 12.5, 25, 50, 100, or 200 µM) for 2 h, transfected with adenovirus-mediated overexpression of HSPB7 for 48 h, or transfected with small interfering RNA of HSPB7 for 48 h followed by exposure to H2O2 (200 µM) for 2 h. Serum concentrations of nonesterified fatty acids and ß-hydroxybutyrate were greater in cows with clinical ketosis, whereas serum concentration of glucose was lower. Compared with healthy cows, the malondialdehyde content was greater but the activity of glutathione peroxidase and superoxide dismutase was lower in adipose tissue of clinically ketotic cows. The abundance of HSPB7 and nuclear factor, erythroid 2 like 2 (NFE2L2) was greater in adipose tissue of clinically ketotic cows. In vitro, H2O2 treatment induced the overproduction of reactive oxygen species and malondialdehyde, and inhibited the activity of antioxidant enzymes glutathione peroxidase and superoxide dismutase in adipocytes from preruminant calves. The low concentration of H2O2 (12.5, 25, and 50 µM) increased the abundance of HSPB7 and NFE2L2, but high concentrations of H2O2 (100 or 200 µM) reduced the abundance of HSPB7 and NFE2L2. The overexpression of HSPB7 improved the H2O2-induced oxidative stress in adipocytes via increasing the abundance of NFE2L2 and its downstream target genes heme oxygenase-1 (HMOX1) and NADH quinone oxidoreductase 1 (NQO1). Knockdown of HSPB7 markedly inhibited the expression of NFE2L2, HMOX1, and NQO1 and further exacerbated H2O2-induced oxidative stress. Overall, these results indicate that activation of the HSPB7-NFE2L2 pathway increases cellular antioxidant capacity, thereby alleviating oxidative stress in bovine adipocytes.

Reconstituted versus dry alfalfa hay in starter feed diets of Holstein dairy calves: Effects on growth performance, nutrient digestibility, and metabolic indications of rumen development.

We investigated the effect of reconstitution of alfalfa hay on starter feed intake, nutrient digestibility, growth performance, rumen fermentation, selected blood metabolites, and health criteria of dairy calves during the pre- and postweaning periods. A total of 20 newborn male Holstein calves (3 d of age; 40.3 ± 1.30 kg of body weight; ±SE) were assigned randomly to 1 of 2 treatments, a starter feed containing either 10% dry (AH) or reconstituted alfalfa hay (RAH), each consisting of 10 calves. Alfalfa hay was reconstituted with water 24 h before feeding to achieve a theoretical dry matter content of 20%. Both starter feeds had the same ingredients and nutrient compositions but differed in their dry matter content (91.2 and 83.8% dry matter for AH and RAH, respectively). Calves were weaned on d 50 and remained on the study until d 70. All calves had free access to fresh and clean drinking water and the starter feed at all times. During the study period, the average maximum temperature-humidity index was 73.8 units, indicating no degree of environmental heat load for dairy calves. Starter feed dry matter intake, total dry matter intake, and body weight (at weaning and at the end of the trial) were unaffected by treatment. Nutrient intake (except for total ether extract intake) increased during the postweaning period compared with the preweaning period. Average daily gain and feed efficiency were unchanged between treatments. Calves had higher average daily gain and skeletal growth during the postweaning period; however, feed efficiency was lower during the post- versus preweaning period. Calves fed RAH gained more hip width and body barrel compared with calves fed AH during the preweaning and all studied periods, respectively. Rectal temperature was similar between treatments, but feeding RAH decreased fecal score and general appearance score during the preweaning period. Apparent total-tract nutrient digestibility was not affected by reconstitution of alfalfa hay; however, reconstitution increased total-tract digestibility of neutral detergent fiber during the postweaning period. Ruminal fluid pH, and concentrations and profile of total volatile fatty acids were unchanged between treatments. Molar concentration of propionate and acetate to propionate ratio increased and decreased, respectively, during the postweaning period. Reconstitution of alfalfa hay did not affect concentrations of glucose, ß-hydroxybutyrate, blood urea N, and albumin, and albumin to globulin ratio during the studied periods; however, reconstitution increased concentration of blood total protein during the overall period. Calves had higher concentrations of blood glucose and globulin during the preweaning and ß-hydroxybutyrate during the postweaning period. Overall, reconstitution of alfalfa hay did not interact with calf phase (pre- vs. postweaning) to affect dry matter intake, growth performance, and metabolic indications of rumen development (measured as ruminal volatile fatty acids and selected blood metabolites), but improved health-related variables (fecal score and general appearance score) during the preweaning period.

Symposium review: Re-evaluation of National Research Council energy estimates in calf starters.

Provision of nutrients in appropriate amounts to meet nutrient requirements for growth, production, and reproduction is the basis for modern animal nutrition. Ration formulation systems predict nutrient requirements based on numerous inputs and then predict nutrient supply based on predicted intake and nutrient content of feeds. Energy systems are used to predict energy supply based on gross caloric content of feeds followed by adjustments for digestion and metabolism of ingested energy. Many models of energy supply use static coefficients of digestibility based on nutrient composition of feed. Other models partition digestion dynamically between ruminal and postruminal digestion but use static estimates of intestinal digestibility to predict energy supplied to the animal. In young calves, both ruminal fermentation and intestinal digestion are underdeveloped; therefore, existing models of energy supply might overestimate the energy available before complete gastrointestinal maturation. In a series of experiments, we reported that total-tract digestion of nutrients changes with advancing age and nutrient intake. Total-tract digestion was measured in calves from 3 to 16 wk of age when fed different amounts and types of milk replacers. Calves were also fed different types of calf starter for ad libitum consumption. Total-tract digestibility of protein, fat, neutral detergent fiber, and nonfiber carbohydrate (NFC) was used to calculate the metabolizable energy (ME) in starter. We used nonlinear regression to estimate the contribution of protein and fat from starter and milk replacer before weaning. Early in life, calculated ME of starter was low and increased with increasing intake of NFC. Cumulative intake of NFC was more highly correlated with changing ME values than other indices, including age, intake of milk replacer, or intake of other nutrients in starter. When calves consumed at least of 15 kg of NFC, ME calculated from digestibility measurements was similar to the ME calculated using National Research Council equations and indicated maturation of gastrointestinal digestion. Our data suggest that intake of NFC is critical to gastrointestinal maturation and the calf's ability to extract energy from calf starter.