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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., (© 2022. The Author(s).)

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., (Copyright © 2021. Published by Elsevier GmbH.)

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 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 β-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.

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., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

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., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

A growing need exists for the development of practical feeding strategies to mitigate methane (CH 4 ) emissions from cattle. Therefore, the objective of this study was to evaluate the influence of milk replacer feeding intensity (MFI) in calves on CH 4 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 CH 4 pre-weaning in wk 9, whereas post-weaning the 20%-MR group tended to have higher levels. Methane emission intensity (CH 4 /BW) was lower pre-weaning in 20%-MR animals but was comparable to the 10%-MR group post-weaning. Methane yield (CH 4 /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 CH 4 emission from calves pre-weaning, although this effect ceases post-weaning., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

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., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

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 β-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., (© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

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., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

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., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

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., (© 2019 Japanese Society of Animal Science.)

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 mm 2 ). 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., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

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 β-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., (© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

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.

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 β-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 β-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., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

This study aimed to explore the roles of microRNAs (miRNAs) in calf rumen development during early life. Rumen tissues were collected from 16 calves (8 at pre-weaning and 8 at post-weaning) for miRNA-sequencing, differential expression (DE), miRNA weighted gene co-expression network (WGCNA) and miRNA-mRNA co-expression analyses. 295 miRNAs were identified. Bta-miR-143, miR-26a, miR-145 and miR-27b were the most abundantly expressed. 122 miRNAs were significantly DE between the pre- and post-weaning periods and the most up- and down-regulated miRNAs were bta-miR-29b and bta-miR-493, respectively. Enrichment analyses of the target genes of DE miRNAs revealed important roles for miRNA in rumen developmental processes, immune system development, protein digestion and processes related to the extracellular matrix. WGCNA indicated that bta-miR-145 and bta-miR-199a-3p are important hub miRNAs in the regulation of these processes. Therefore, bta-miR-143, miR-29b, miR-145, miR-493, miR-26a and miR-199 family members might be key regulators of calf rumen development during early life., (Crown Copyright © 2018. Published by Elsevier Inc. All rights reserved.)

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 H 2 O 2 (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 H 2 O 2 (200 μM) for 2 h. Serum concentrations of nonesterified fatty acids and β-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, H 2 O 2 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 H 2 O 2 (12.5, 25, and 50 μM) increased the abundance of HSPB7 and NFE2L2, but high concentrations of H 2 O 2 (100 or 200 μM) reduced the abundance of HSPB7 and NFE2L2. The overexpression of HSPB7 improved the H 2 O 2 -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 H 2 O 2 -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., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

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., Competing Interests: This study was partially funded by DSM (Produtos Nutricionais Brasil S.A.), but this does not alter our adherence to PLOS ONE policies on sharing data and materials.

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, β-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 β-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., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

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., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Microbial colonisation in the forestomach of a ruminant is one of the most crucial factors in determining many of its physiological developments and digestive capabilities. The present study attempts to identify establishment pattern of microbes in relation to food, age and rumen development in the buffalo calves at every fortnight interval from birth to 6 months of age, followed by every month till animals became 1 year of age. Diversity study based on 16S rRNA gene sequencing identified rapidly changing bacterial population during initial 60 days of life, which got assemblage as rumen became physiologically mature with increasing age of animals. A lactate fermenting aerobic to facultative anaerobic genera found during initial 30 days of life were expeditiously replaced by strict anaerobic cellulolytic bacterial population with increasing age. The study confirms that initial colonisation mainly depends on the oral cavity and skin of the mother, followed by the surrounding environment and feed offered, which is reversed in order once animal gets older. Some of the well-described genera based on culture-dependent studies like Ruminococcus spp. were found to be in lesser proportion suggesting an additional role of other microbes or niche in cellulose degradation. We report the presence of Porphyromonas spp. and Mannheimia glucosidal for the first time in bovine infants.

In order to investigate the effect of rearing systems on growth and rumen development in Malabari male kids, 14 pre-weaned Malabari male kids of uniform morphological characters were randomly divided into two equal groups as T 1 and T 2 . Both the groups were reared intensively for 12 weeks whereby kids under T 1 group were allowed to suckle their dams and provided green grass ad libitum. The kids under T 2 were weaned at the age of 7 days and provided formulated semi-solid broiler goat concentrate diet through the feeding bottle and were not offered any grass/roughage. Parameters like live weight gain, daily weight gain and body measurements were studied to evaluate the growth performance. However, gastrointestinal tract morphometric studies and qualitative morphological analysis of rumen papillae were undertaken to measure the rumen development. The daily weight gain of kids under T 2 was significantly (P ≤ 0.01) higher than the daily weight gain of kids under T 1 . Also, the body measurements like height at withers, heart girth and body length were significantly (P ≤ 0.01) higher in T 2 than in T 1 . The rumen and abomasum were comparatively more developed in T 2 than in T 1 . The morphology of rumen papillae in both groups was different in size, shape and colour. The length, width, density and surface area of rumen papillae among kids in T 2 were significantly (P ≤ 0.01) higher than those kids in T 1 . The results of the present study indicated that the feeding of semi-solid broiler goat concentrate diet improved growth performance and early rumen development in kids.

The length and density of rumen papillae starts to increase during weaning and growth of ruminants. This significant development increases the intraruminal surface area and the efficiency of VFA (acetate, propionate, butyrate, etc.) uptake. Thus, it is important to investigate the factors controlling the growth and development of rumen papillae during weaning. This study aimed to compare the transcriptomes of rumen papillae in suckling and weaned calves. Total RNA was extracted from the rumen papillae of 10 male Japanese Black calves (5 suckling calves, 5 wk old; 5 weaned calves, 15 wk old) and used in RNA-sequencing. Transcript abundance was estimated and differentially expressed genes were identified and these data were then used in Ingenuity Pathway Analysis (IPA) to predict the major canonical pathways and upstream regulators. Among the 871 differentially expressed genes screened by IPA, 466 genes were upregulated and 405 were downregulated in the weaned group. Canonical pathway analysis showed that "atherosclerosis" was the most significant pathway, and "tretinoin," a derivative of vitamin A, was predicted as the most active upstream regulator during weaning. Analyses also predicted IgG, lipopolysaccharides, and tumor-necrosis factor-α as regulators of the microbe-epithelium interaction that activates rumen-related immune responses. The functional category and the up-regulators found in this study provide a valuable resource for studying new candidate genes related to the proliferation and development of rumen papillae from suckling to weaning Japanese Black calves.

The development of the gastrointestinal tract in newborn calves is essential for sufficient nutrient uptake. An intensive milk feeding during the neonatal period may impair the rumen development in calves. The aim of this study was to investigate effects of milk replacer (MR) feeding in unlimited amounts for the first 5 wk of age on the gastrointestinal growth and development in preruminant calves at wk 9 of age. Twenty-eight newborn Holstein and Holstein × Charolais crossbred calves (19 male and 9 female) were fed MR ad libitum (ADLIB) or in restricted amounts (6 L per day; RES) until wk 5 of age. Thereafter, the MR intake of ADLIB was gradually reduced at wk 6 and 7, and all calves received 6 L of MR per day until wk 9 of age. In wk 9, calves were slaughtered and carcass and organ weight as well as rumen papilla size in the atrium, ventral sac, and ventral blind sac, and villus size of the mucosa in the small intestine (duodenum; proximal, mid, and distal jejunum; and ileum) were determined. The expression of mRNA associated with the local insulin-like growth factor (IGF) system was measured in the rumen epithelium. Ad libitum MR feeding increased MR intake and growth in ADLIB without influencing concentrate intake compared with RES. Carcass weight in wk 9 was greater in ADLIB than in RES. The density of the rumen papillae in the atrium and ventral blind sac was greater in RES than in ADLIB calves, but surface area of the epithelium was not different between groups in the investigated regions of the rumen. The mRNA abundance of IGF1 in the atrium tended to be greater and the IGFR1 mRNA abundance in the ventral sac tended to be lower in the ADLIB than in the RES feeding group. The rumen pH and volatile fatty acid concentrations were not affected by MR feeding intensity. In mid-jejunum, villus circumference was greater in ADLIB than in RES calves. In the distal jejunum, villus surface area and the villus height/crypt depth ratio were greater and the villus circumference and height tended to be greater, whereas crypt depth was smaller in ADLIB than in RES calves. The findings from this study indicate that ad libitum MR feeding for 5 wk of age followed by its gradual reduction promotes growth performance without any negative influence on gastrointestinal growth and development in dairy calves at 9 wk of age., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The importance of the use of exogenous butyrate in calves' diets is due to its role as a factor stimulating the functional development of ruminal epithelium and improving calf performance during the transition from preruminant to ruminant status. Our review will first present results related to effects of the administration of butyrate in calves' diets on the development of ruminal epithelium toward a more effective absorption and metabolism of fermentation products from the rumen. The introduction of sodium butyrate at a level of about 0.3% of diet dry matter is accompanied by an increase to 35% in butyrate concentration in the rumen of 33-day-old calves. Mutual reliance between an enhanced ruminal concentration of butyrate and the activities of transcription factors, genes and proteins involved in cell proliferation, ketogenesis and the maintenance of cell pH homeostasis in the ruminal epithelial cells has been clearly confirmed in many experiments. Second, the review presents results related to the effects of the introduction of butyrate salts in the diet on calf performance. Of 11 studies a positive effect was found in six; five of these were obtained from the calves that started receiving butyrate supplement at a level of about 0.3% diet dry matter from the age of 3 to 5 days. Results indicate that when a supplement is given to calves soon after birth the functional development of ruminal epithelium in cooperation with the endocrine and digestion systems is transferred into improving the efficiency of rearing. There have been no studies on the effects of greater amounts of butyrate salts in milk replacer; butyrate constitutes about 1.2% of the whole cow's milk dry matter. In older calves, when butyrate administration is provided as a component of a starter concentrate at the increasing inclusion rate from 0.3% to 3.0%, the practical effect in calf performance relates to the risk of depression of rumen pH below 5.5 and accompanying disruption of the organization of the ruminal epithelial tissue. The higher risk is noted in calves received starter with substantial content of a rapidly degradable starch. At present, the insufficient number of positive results confirming the beneficial effect of butyrate supplements in terms of an improvement in performance does not allow their recommendation for use in the practical feeding of calves.

We previously reported 2 experiments with rumen-cannulated Holstein-Friesian dairy cows showing that during the transition period, rumen papillae surface area, and fractional absorption rate of volatile fatty acids (VFA) increase after calving. However, supplemental concentrate during the dry period and rate of increase of concentrate allowance during lactation affected papillae surface area, but not VFA absorption. Here we report the changes in gene and protein expression in rumen papillae related to tissue growth and VFA utilization. The lactation experiment treatment consisted of a rapid [RAP; 1.0 kg of dry matter (DM)/d; n = 6] or gradual (GRAD; 0.25 kg of DM/d; n = 6) increase of concentrate allowance (up to 10.9 kg of DM/d), starting at 4 d postpartum (pp). The dry period experiment treatment consisted of 3.0 kg of DM/d of concentrate (n = 4) or no concentrate (n = 5) during the last 28 d of the dry period. Real-time quantitative PCR analysis of rumen papillae showed that the expression of apoptosis-related genes was neither affected by day nor its interaction with treatment for both experiments. Expression of epithelial transporter genes was not affected by day or treatment in the lactation experiment, except for NBC1. In the dry period experiment, expression of MCT1, NBC1, DRA, NHE2, NHE3, and UT-B generally decreased after calving. A day and treatment interaction was observed for ATP1A1 in the dry period experiment, with greater expression at 18 and 8 d antepartum for concentrate than no concentrate. Generally, expression of VFA metabolism-related genes was not affected by day or its interaction with treatment. In the lactation experiment, immunoblotting of 5 selected genes showed that protein expression of DRA and PCCA was greater at 16 d pp compared with 3 and 44 d pp. Expression of NHE2 was greater, and that of ATP1A1 lower, at 16 and 44 d pp compared with 3 d pp, suggesting alterations in intracellular pH regulation and sodium homeostasis. Both MCT1 and PCCA protein were upregulated by RAP from 3 to 16 d pp, indicating modulations in VFA metabolism. Our data suggests that VFA absorption and metabolic capacity changed little per unit of surface area during the transition period, and suggests that a change in mitosis rate rather than apoptosis rate is associated with the increased ruminal VFA production, resulting in tissue growth. A significant but weak correlation between the examined gene and protein expression levels was observed only for PCCA, indicating that care must be taken when interpreting results obtained at either level., (Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Compensatory growth (CG) is utilised worldwide in beef production systems as a management approach to reduce feed costs. However the underlying biology regulating the expression of CG remains to be fully elucidated. The objective of this study was to examine the effect of dietary restriction and subsequent re-alimentation induced CG on the global gene expression profile of ruminal epithelial papillae. Holstein Friesian bulls (n = 60) were assigned to one of two groups: restricted feed allowance (RES; n = 30) for 125 days (Period 1) followed by ad libitum access to feed for 55 days (Period 2) or (ii) ad libitum access to feed throughout (ADLIB; n = 30). At the end of each period, 15 animals from each treatment were slaughtered and rumen papillae harvested. mRNA was isolated from all papillae samples collected. cDNA libraries were then prepared and sequenced. Resultant reads were subsequently analysed bioinformatically and differentially expressed genes (DEGs) are defined as having a Benjamini-Hochberg P value of <0.05. During re-alimentation in Period 2, RES animals displayed CG, growing at 1.8 times the rate of their ADLIB contemporary animals in Period 2 (P < 0.001). At the end of Period 1, 64 DEGs were identified between RES and ADLIB, with only one DEG identified at the end of Period 2. When analysed within RES treatment (RES, Period 2 v Period 1), 411 DEGs were evident. Genes identified as differentially expressed in response to both dietary restriction and subsequent CG included those involved in processes such as cellular interactions and transport, protein folding and gene expression, as well as immune response. This study provides an insight into the molecular mechanisms underlying the expression of CG in rumen papillae of cattle; however the results suggest that the role of the ruminal epithelium in supporting overall animal CG may have declined by day 55 of re-alimentation.

Isobutyrate supplements could improve rumen development by increasing ruminal fermentation products, especially butyrate, and then promote the growth performance of calves. The objective of this study was to evaluate the effects of isobutyrate supplementation on growth performance, rumen development, blood metabolites and hormone secretion in pre- and post-weaned dairy calves. In total, 56 Chinese Holstein male calves with 30 days of age and 72.9±1.43 kg of BW, blocked by days of age and BW, were assigned to four groups in a randomized block design. The treatments were as follows: control, low-isobutyrate, moderate-isobutyrate and high-isobutyrate with 0, 0.03, 0.06 and 0.09 g isobutyrate/kg BW per calf per day, respectively. Supplemental isobutyrate was hand-mixed into milk of pre-weaned calves and the concentrate portion of post-weaned calves. The study consisted of 10 days of an adaptation period and a 50-day sampling period. Calves were weaned at 60 days of age. Seven calves were chosen from each treatment at random and slaughtered at 45 and 90 days of age. BW, dry matter (DM) intake and stomach weight were measured, samples of ruminal tissues and blood were determined. For pre- and post-weaned calves, DM intake and average daily gain increased linearly (P<0.05), but feed conversion ratio decreased linearly (P<0.05) with increasing isobutyrate supplementation. Total stomach weight and the ratio of rumen weight to total stomach weight tended to increase (P=0.073) for pre-weaned calves and increased linearly (P=0.021) for post-weaned calves, whereas the ratio of abomasum weight to total stomach weight was not affected for pre-weaned calves and decreased linearly (P<0.05) for post-weaned calves with increasing isobutyrate supplementation. Both length and width of rumen papillae tended to increase linearly for pre-weaned calves, but increased linearly (P<0.05) for post-weaned calves with increasing isobutyrate supplementation. The relative expression of messenger RNA for growth hormone (GH) receptor and 3-hydroxy-3-methylglutaryl-CoA synthase 1 in rumen mucosa increased linearly (P<0.05) for pre- and post-weaned calves with increasing isobutyrate supplementation. Blood concentrations of glucose, acetoacetate, β-hydroxybutyrate, GH and IGF-1 increased linearly (P<0.05) for pre- and post-weaned calves, whereas blood concentration of insulin decreased linearly with increasing isobutyrate supplementation. The present results indicated that isobutyrate promoted growth of calves by improving rumen development and its ketogenesis in a dose-dependent manner.

This study was performed to investigate the initial colonization of metabolically active methanogens and subsequent changes in four fractions: the rumen solid-phase (RS), liquid-phase (RL), protozoa-associated (RP), and epithelium-associated (RE) from 1 to 60 d after birth, and manipulate methanogen community by early weaning on 40 d and supplementing rhubarb from 40 to 60 d in black goats. The RNA-based real-time quantitative PCR and 16S rRNA amplicon sequencing were employed to indicate the metabolically active methanogens. Results showed that active methanogens colonized in RL and RE on 1 d after birth. RP and RE contained the highest and lowest density of methanogens, respectively. Methanobrevibacter, Candidatus Methanomethylophilus, and Methanosphaera were the top three genera. The methanogen communities before weaning differed from those post weaning and the structure of the methanogen community in RE was distinct from those in the other three fractions. The discrepancies in the distribution of methanogens across four fractions, and various fluctuations in abundances among four fractions according to age were observed. The addition of rhubarb significantly (P < 0.05) reduced the abundances of Methanimicrococcus spp. in four fractions on 50 d, but did not change the methanogen community composition on 60 d.

The rumen is lined on the luminal side by a stratified squamous epithelium that is responsible for not only absorption, but also transport, extensive short-chain fatty acid (SCFA) metabolism and protection. Butyrate has been demonstrated to initiate the differentiation of the tissue following introduction of solid feed to the weaning neonate as well as affecting the metabolism of other nutrients and absorption of nutrients in in vitro experiments. The objective of the present study was to validate expression stability of eight putative reference genes bovine rumen, considering the intrinsic heterogeneity of bovine rumen with regard to different luminal characteristics due to direct infusion of butyrate to double the intra-ruminal content of the rumen liquor. Our focus was on identifying stable reference genes which are suitable to normalize real-time RT-qPCR experiments from rumen samples collected from clinical assays, irrespective of localization within the organ and the across physiological state. The most stably expressed genes included: ACTB, UXT, DBNDD2, RPS9, DDX54 and HMBS. Their high stability values suggest these reference genes will facilitate better evaluation of variation of across an array of conditions including: localization within the rumen, differences among cattle fed an array of rations, as well as response to development in the weaning animal. Moreover, we anticipate these reference genes may be useful for expression studies in other ruminants.

Feeding of neonates with artificial milk formulas is a popular trend toward early weaning of newborn dairy calves. These milk replacers (MR) should accelerate the rumen development, determining early solid feed intake and leading to better performances in cattle. Previous research demonstrated that sodium butyrate supplementation in MR can affect both small intestine and rumen development. Also acetate and propionate showed similar properties, while only a few studies indicate some potential benefit of monoglycerides on gut functions. The present study is aimed to determine the effect of the supplementation of a blend containing short and medium chain fatty acids monoglycerides (SMCFA) in milk replacer on rumen papillae development and growth performances in weaning calves. Twenty bull calves (about 2 weeks old, weighing around 43kg) were randomly allocated into two groups: control (C) and treated (T). Besides MR and starter diet, the latter offered at libitum, T calves received 0.2% SMCFA in MR. Animals were slaughtered after 56 days from the beginning of the trial. No difference was found between groups either in growth performances or in mean number of papillae/cm(2) of mucosa, total surface of papillae (mm(2))/cm(2) of mucosa or papillary size. In both groups, the morphology of the rumen epithelium was typical of parakeratosis. The cells of the stratum spinosum were directly transformed into swollen, ovoid, still nucleated keratinocytes, particularly at the papillary tip, probably as a result of unphysiological osmolarities caused by high concentrate intake. Degenerated squamous horn cells covered the "balloon like" cells forming several layers, particularly in the places of the rumen mucosa more protected from an abrasive action of solid feed. This was more evident in C animals. The squamous cells covering the papillary tip showed cytoplasmic protrusion, representing remains of the attachment sites of desmosomes, which increased the total absorptive surface and were more numerous and higher in T compared to C animals. It might be hypothesized that SMCFA supplementation in MR could better regulate epithelial cell proliferation and probably have an "emollient effect" leading to an easier "peeling" that might increase efficiency for nutrient transport across the epithelium., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Recent research suggests that circulating β-hydroxybutyrate (BHB) levels may be a meaningful indicator of grain intake and rumen development in pre-ruminant calves. As such, BHB levels may be a surrogate measure of rumen function to contribute to minimal weaning stress during the transition from liquid to solid feed. The primary objective of this study was to determine the optimal cut-point of circulating BHB levels that would be predictive of sufficient grain intake and rumen development for a successful transition from liquid to solid feed at the time of weaning. An additional objective was to validate the Precision Xtra (Abbott Diabetes Care, Abingdon, UK) calf-side test for determination of BHB in whole blood in calves around weaning, as compared with the gold standard laboratory method. A total of 20 Holstein female calves were randomly assigned at birth to be weaned at 6 wk (n=10) or 8 wk (n=10) of age. Milk replacer (mixed at 150 g/L) was offered at 1.2kg/calf per d in 2 meals until a 1-wk step down, when milk meals were reduced by 50% 1 wk before weaning. Daily measurements included the intakes of starter grain, chopped straw, and water. Weekly measurements included body weight and blood BHB, until 70 d of life. To assess digestive tract development, rumen fluid samples were taken before and after weaning (d 35, 49, and 63) and analyzed for ruminal short-chain fatty acids. Whole blood was collected by jugular venipuncture, and BHB was determined by the Precision Xtra test at calf-side. In addition, serum was separated from a clotted sample, frozen, and stored until laboratory analysis was performed. Laboratory BHB results were correlated with both the Precision Xtra test (r=0.95) and starter intake over 1 d (r=0.89), a 3-d average (r=0.90), and a 7-d average (r=0.90). Additionally, laboratory BHB results were associated with total ruminal volatile fatty acids (r=0.82), ruminal butyrate (r=0.77), and body weight (r=0.69). A receiver operating characteristic curve was created to plot the true positive rate against the false positive rate at 10μmol/L BHB intervals to determine the optimal cut-point of circulating BHB that is predictive of an average starter intake of 1,000 g/d over a 3-d period. The optimal combination of sensitivity (95.7%) and specificity (96.1%) was at 100µmol of BHB/L of blood. A value of 0.2 mmol/L on the Precision Xtra test yielded a sensitivity and specificity of 84.0 and 97.2%, respectively, over the 3-d average period of starter intake. These results show considerable promise for use of the Precision Xtra whole blood BHB test in the decision-making process of determining sufficient starter grain intake and rumen development for a successful transition from liquid to solid feed, and indicate that this test conducted at calf-side is highly accurate., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Early consumption of starter feed promotes rumen development in lambs. We examined rumen development in lambs fed starter feed for 5 weeks using histological and biochemical analyses and by performing high-throughput sequencing in rumen tissues. Additionally, rumen contents of starter feed-fed lambs were compared to those of breast milk-fed controls. Our physiological and biochemical findings revealed that early starter consumption facilitated rumen development, changed the pattern of ruminal fermentation, and increased the amylase and carboxymethylcellulase activities of rumen micro-organisms. RNA-seq analysis revealed 225 differentially expressed genes between the rumens of breast milk- and starter feed-fed lambs. These DEGs were involved in many metabolic pathways, particularly lipid and carbohydrate metabolism, and included HMGCL and HMGCS2. Sequencing analysis of 16S rRNA genes revealed that ruminal bacterial communities were more diverse in breast milk-than in starter feed-fed lambs, and each group had a distinct microbiota. We conclude that early starter feeding is beneficial to rumen development and physiological function in lambs. The underlying mechanism may involve the stimulation of ruminal ketogenesis and butanoate metabolism via HMGCL and HMGCS2 combined with changes in the fermentation type induced by ruminal microbiota. Overall, this study provides insights into the molecular mechanisms of rumen development in sheep.

This study investigated the effects of rumen-protected methionine (RPM) and rumen-protected choline (RPC) on energy balance, postpartum lactation performance, antioxidant capacity and immune response in transition dairy cows. Forty-eight multiparous transition cows were matched and divided into four groups: control, 15 g/d RPC, 15 g/d RPM or 15 g/d RPC + 15 g/d RPM. Diet samples were collected daily before feeding, and blood samples were collected weekly from the jugular vein before morning feeding from 21 days prepartum to 21 days postpartum. Postpartum dry matter intake (DMI) was increased by both additives (P < 0.05), and energy balance values in supplemented cows were improved after parturition (P < 0.05). Both RPC and RPM decreased the plasma concentrations of non-esterified fatty acids (NEFA), β-hydroxybutyric acid (BHBA), total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) (P < 0.05), but increased the plasma levels of glucose, very-low-density lipoprotein (VLDL) and apolipoprotein B100 (ApoB 100, P < 0.05). The supplements improved milk production (P < 0.05), and increased (P < 0.05) or tended to increase (0.05 < P < 0.10) the contents of milk fat and protein. The post-ruminal choline and methionine elevated the blood antioxidant status, as indicated by total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px) activity and the vitamin E concentration (P < 0.05), and reduced the plasma malondialdehyde (MDA) level (P < 0.05). Furthermore, RPM and RPC elevated the plasma interleukin 2 (IL-2) concentration and the CD4+/CD8+ T lymphocyte ratio in peripheral blood (P < 0.05). Alternatively, the levels of tumor necrosis factor-α (TNF-α) and IL-6 were decreased by RPM and RPC (P < 0.05). Overall, the regulatory responses of RPC and RPM were highly correlated with time and were more effective in the postpartum cows. The results demonstrated that dietary supplementation with RPC and RPM promoted energy balance by increasing postpartal DMI and regulating hepatic lipid metabolism, improved postpartum lactation performance and enhanced antioxidant capacity and immune function of transition dairy cows.

The aim of this study was to evaluate the effects of Saccharomyces cerevisiae fermentation products (SCFP) in the calf starter and milk on ruminal fermentation, gastrointestinal morphology, and microbial community in the first 56 d of life. Thirty Holstein bull calves were randomly assigned to 1 of 3 groups: a texturized calf starter containing 0 (CON), 0.5, or 1% SCFP (XPC, Diamond V, Cedar Rapids, IA) of dry matter from d 4 to 56. In addition, the XPC-supplemented calves were fed with 1 g/d SCFP (SmartCare, Diamond V, Cedar Rapids, IA) in milk from d 2 to 30. All calves were fed 4 L of colostrum within 1 h of birth and were subsequently fed milk twice daily until weaned on d 56. Rumen fluid was collected by an esophageal tube 4 h after the morning feeding on d 28 and 56 to determine ruminal pH, ammonia-N, and volatile fatty acids concentrations. On d 56, 15 (5 per treatment) calves were harvested and slaughter weight, gastrointestinal morphology parameters, and bacteria community were recorded. Papilla length, width, and surface area were measured from 5 locations within the rumen. Villus height, width, surface area, crypt depth, and villus height-to-crypt depth ratio were measured in the duodenum, jejunum, and ileum. Next-generation sequencing technology was used to test the microbial community of the rumen and duodenum samples on d 28 and 56. Data were analyzed by MIXED procedure in SAS (SAS Institute Inc., Cary, NC) with contrast statements to declare CON versus all SCFP and 0.5 versus 1% SCFP in starter grains. Ruminal pH, ammonia-N, and total volatile fatty acids were not altered by SCFP. However, the supplemented groups exhibited higher ruminal butyrate concentrations coinciding with higher Butyrivibrio and lower Prevotella richness than CON group. Supplementation of SCFP increased papilla length in the rumen. In the small intestine, SCFP reduced crypt depth of jejunum, and increased villus height-to-crypt depth ratio in all segments of the small intestine, especially when supplemented at a higher dosage in the starter. In conclusion, Saccharomyces cerevisiae fermentation products improved gastrointestinal morphology, possibly due to increased Butyrivibrio and decreased Prevotella richness of the rumen fluid, which resulted in an increase in butyrate production, and the effect was slightly greater with the higher dosage of SCFP in the starter., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

This study aimed to identify the genes associated with the development of the rumen epithelium by screening for candidate genes by digital differential display (DDD) in silico. Using DDD in NCBI's UniGene database, expressed sequence tag (EST)-based gene expression profiles were analyzed in rumen, reticulum, omasum, abomasum and other tissues in cattle. One hundred and ten candidate genes with high expression in the rumen were derived from a library of all tissues. The expression levels of 11 genes in all candidate genes were analyzed in the rumen, reticulum, omasum and abomasum of nine Japanese Black male calves (5-week-old pre-weaning: n = 3; 15-week-old weaned calves: n = 6). Among the 11 genes, only 3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2), aldo-keto reductase family 1, member C1-like (AKR1C1), and fatty acid binding protein 3 (FABP3) showed significant changes in the levels of gene expression in the rumen between the pre- and post-weaning of calves. These results indicate that DDD analysis in silico can be useful for screening candidate genes related to rumen development, and that the changes in expression levels of three genes in the rumen may have been caused by weaning, aging or both., (© 2015 Japanese Society of Animal Science.)

Calves are born with a physically and metabolically underdeveloped rumen and initially rely on milk to meet nutrient demands for maintenance and growth. Initiation of solid feed consumption, acquisition of anaerobic microbes, establishment of rumen fermentation, expansion of rumen in volume, differentiation and growth of papillae, development of absorption and metabolic pathways, maturation of salivary apparatus and development of rumination behavior are all needed as the calf shifts from dependence on milk to solid feed. In nature and some production systems (e.g., most beef calves), young ruminants obtain nutrients from milk and fresh forages. In intensive dairying, calves are typically fed restricted amounts of milk and weaned onto starter feeds. Here we review the empirical work on the role of feeding and management during the transition from milk to solid feed in establishing the rumen ecosystem, rumen fermentation, rumen development, rumination behavior, and growth of dairy calves. In recent years, several studies have illustrated the benefits of feeding more milk and group rearing of dairy calves to take advantage of social facilitation (e.g., housing with peers or dam), and this review also examines the role of solid feed on rumen development and growth of calves fed large quantities of milk and reared under different housing situations. We conclude that the provision of high-starch and low-fiber starter feeds may negatively affect rumen development and that forage supplementation is beneficial for promoting development of the gut and rumination behavior in young calves. It is important to note that both the physical form of starter diets and their nutritional composition affect various aspects of development in calves. Further research is warranted to identify an optimal balance between physically effective fiber and readily degradable carbohydrates in starter diets to support development of a healthy gut and rumen, rumination behavior, and growth in young calves., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

This study aims to determine the effects of alfalfa supplementation on the pre- and postweaning performance, rumen development, and feed transition in starter diet-fed lambs. Six of 66 male Hu lambs were slaughtered at the age of 10 d to serve as a control. The other 60 lambs were randomly allocated to 2 dietary treatments: milk replacer and starter pellets without (STA) or with free-choice chopped alfalfa (S-ALF). The animals were offered 300 g/d of the concentrate mixture and had free access to alfalfa after weaning at the end of wk 4 (age 38 d). The alfalfa inclusion in the S-ALF group tended to increase the starter intake before weaning, significantly increased the concentrate intake soon after weaning ( < 0.05), and increased the BW ( < 0.01) and ADG ( < 0.10) in pre- and postweaning lambs. The S-ALF group had heavier carcasses ( < 0.05), rumens ( < 0.05), reticula ( < 0.05), omasums ( < 0.10), abomasums ( < 0.05), and visceral organs ( < 0.10) than the STA lambs after weaning. Alfalfa supplementation increased ( < 0.05) the rumen papillae length and the ratio of the duodenal villus height to the crypt depth; it also decreased ( < 0.05) the concentration and molar proportion of propionate in wk 1 and 5. The STA lambs had higher ( < 0.01) blood concentrations of globulin and blood urea nitrogen and lower β-hydroxybutyrate after weaning. The STA group also had a higher incidence of feed plaque. From the above results, we infer that the free-choice addition of chopped alfalfa to starter diets is beneficial to rumen development, relieves weaning stress, and improves the performance of lambs.

Thirty calves were randomly assigned to two treatments and fed until weaning [42 days (d) of age]. Treatments were a control group (n = 15), which did not receive Megasphaera elsdenii (Me0) and a M. elsdenii group, which received a 50-ml oral dose of M. elsdenii NCIMB 41125 (10(8) CFU/ml) at day 14 day of age (Me14). Calves were given colostrum for the first 3 day followed by limited whole milk feeding. A commercial calf starter was offered ad libitum starting at day 4 until the end of the study. Fresh water was available throughout the study. Feed intake and growth were measured. Blood samples were collected via jugular venipuncture to determine β-hydroxybutyrate (BHBA) concentrations. Fourteen male calves (seven per group) were euthanised on day 42 and digestive tracts harvested. Reticulo-rumen weight was determined and rumen tissue samples collected from the cranial and caudal sacs of the ventral and dorsal portions of the rumen for measurements of papillae length, papillae width and rumen wall thickness. Dosing with M. elsdenii NCIMB 41125 improved starter dry matter intake (DMI), weaning body weight (BW) and tended to improve average daily gain. Calves in Me14 group had greater plasma BHBA concentration than Me0-calves during the last 3 weeks of the trial and had at day 42 greater reticulo-rumen weight, papillae width and papillae density compared to Me0. No differences in rumen wall thickness or papillae length were observed between the two groups. Total volatile fatty acids, acetate and propionate production did not differ between treatments, but butyrate production was greater in Me14 than Me0. Dosing M. elsdenii NCIMB 41125 showed benefit for calves with improved feed intake and rumen development suggesting increased epithelium metabolism and improved absorption of digestive end products., (Journal of Animal Physiology and Animal Nutrition © 2015 Blackwell Verlag GmbH.)

Inclusion of forage and its physical form in starter may affect rumen development, average daily gain (ADG), and dry matter intake (DMI) of dairy calves. To evaluate the effects of forage and its physical form (chopped vs. pelleted) on growth of calves under a high milk feeding regimen, 32 Holstein calves (38.8±1.1kg) were assigned at birth to 1 of 3 treatments in a completely randomized block design. Dietary treatments (% of dry matter) were (1) 100% semi-texturized starter (CON); (2) 90% semi-texturized starter + 10% chopped alfalfa hay (mean particle size=5.4mm) as a total mixed ration (TMR; CH); and (3) 90% semi-texturized starter + 10% pelleted alfalfa (mean=5.8mm) hay as a TMR (PH). Data were subjected to mixed model analysis with contrasts used to evaluate effect of forage inclusion. Calves were weaned at 76 d of age and the experiment finished 2 wk after weaning. Individual milk and solid feed consumption were recorded daily. Solid feed consumption and ADG increased as age increased (effect of week), but neither forage inclusion nor physical form of forage affected these variables pre- or postweaning. Plasma urea N was affected by treatments such that the CON group had a lower concentration than forage-fed groups. Forage inclusion, but not physical form, resulted in increased total protein in plasma. Although days with elevated rectal temperature, fecal score, and general appearance were not affected by dietary treatments, calves fed alfalfa hay during the first month of life had fewer days with respiratory issues, regardless of physical form of hay. We concluded that provision of forage does have some beneficial effects in calves fed large amounts of milk replacer, but pelleted alfalfa hay did not result in any improvement in calf performance or health., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The aim of this study was to assess the effects of particle size (PS) of alfalfa hay on growth characteristics and rumen development in dairy calves at two levels of alfalfa supplementation. Fifty newborn dairy calves (42.7 ± 2.2 kg BW) were used in a 2 × 2 factorial arrangement with the factors supplementation level (low, 8%; or high, 16% on DM basis) and PS (medium, 2.92 mm; or long, 5.04 mm as geometrical means) of alfalfa hay. In addition, a control group without alfalfa hay was used. Hence, treatments were: control (C); low level with medium PS (LM); low level with long PS (LL); high level with medium PS (HM) or high level with long PS (HL). Growth performance of alfalfa-fed calves did not differ from control calves, but alfalfa supplementation decreased corneum thickness of the rumen wall. In alfalfa-fed calves, post-weaning starter intake was greater for LL calves than for LM calves. During the entire rearing period, starter intake was 26-32% higher for LL and HM calves than for LM calves. Pre-weaning average daily gain was higher for LL and HM calves than for HL calves, but this effect was not persistent over the entire rearing period. Final body weight decreased from 86 to 79 kg when the level of long PS alfalfa hay increased from 8 to 16%, but increased from 78 to 87 kg when the level of medium PS alfalfa increased from 8 to 16%. Regardless of PS and level, morphometric characteristics of rumen wall were generally similar among alfalfa feeding groups, but corneum thickness decreased from 8.7 to 6.1 μm with greater PS at the low level. These results indicate that adequate, but not excessive, physical stimulation is required for appropriate rumen development and growth performance of dairy calves., (Journal of Animal Physiology and Animal Nutrition © 2014 Blackwell Verlag GmbH.)

Understanding of the colonization process of epithelial bacteria attached to the rumen tissue during rumen development is very limited. Ruminal epithelial bacterial colonization is of great significance for the relationship between the microbiota and the host and can influence the early development and health of the host. MiSeq sequencing of 16S rRNA genes and quantitative real-time PCR (qPCR) were applied to characterize ruminal epithelial bacterial diversity during rumen development in this study. Seventeen goat kids were selected to reflect the no-rumination (0 and 7 days), transition (28 and 42 days), and rumination (70 days) phases of animal development. Alpha diversity indices (operational taxonomic unit [OTU] numbers, Chao estimate, and Shannon index) increased (P < 0.01) with age, and principal coordinate analysis (PCoA) revealed that the samples clustered together according to age group. Phylogenetic analysis revealed that Proteobacteria, Firmicutes, and Bacteroidetes were detected as the dominant phyla regardless of the age group, and the abundance of Proteobacteria declined quadratically with age (P < 0.001), while the abundances of Bacteroidetes (P = 0.088) and Firmicutes (P = 0.009) increased with age. At the genus level, Escherichia (80.79%) dominated at day zero, while Prevotella, Butyrivibrio, and Campylobacter surged (linearly; P < 0.01) in abundance at 42 and 70 days. qPCR showed that the total copy number of epithelial bacteria increased linearly (P = 0.013) with age. In addition, the abundances of the genera Butyrivibrio, Campylobacter, and Desulfobulbus were positively correlated with rumen weight, rumen papilla length, ruminal ammonia and total volatile fatty acid concentrations, and activities of carboxymethylcellulase (CMCase) and xylanase. Taking the data together, colonization by ruminal epithelial bacteria is age related (achieved at 2 months) and might participate in the anatomic and functional development of the rumen., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

The effect of fermented apple pomace (FAP) on animal health, antioxidant activity (AA), hematic biometry (HBm) and the development of ruminal epithelium were investigated in a study with 24 finishing lambs (BW = 25.4 ± 3.3 Kg). Lambs were grouped by sex (12 male and 12 female) and fed (n = 6 per group of treatment) a basal fattening diet (Control diet, T1) or the basal diet supplemented to include 10.91% of fermented apple pomace (FAP diet, T2). The animals were kept 56 d in individual metabolic cages, with ad libitum access to water and feed. Two blood samples were collected from each animal on d 0, 28, and 56 to determine AA in plasma and hematic biometry (HBm). Four samples of ruminal tissue were taken postmortem to evaluate the development of ruminal epithelium based on the length (LP) and width (WP) of papillae. AA and HBm data were analyzed with a mixed model (fixed effects: diet, sampling, sex, and their interaction; using the experimental unit nested in the effect of the diet as the random effect). LP and WP were analyzed with a hierarchical model, as simple and nested effects in the sampling site, where the fixed effects were the diet and the sex of the animal and their interaction. There was an effect of diet on AA, which was higher (P < 0.06) in T2 vs. T1 at 56 d (24.34 vs. 21.79 mM Fe2). Leukocytes increased (P < 0.05) from 7.52*10(3) ± 1.29*10/(3)μL to 9.14*10(3) ± 1.24*10(3)/μL in all the animals in the experiment, with a marked increased (P < 0.05) at 28 d after beginning of the feeding period, with values within the normal range for this species and without effect of the diet (P > 0.05) for the other indicators of HBm. Males' LP was higher in T2 than in T1 (P < 0.05). It was concluded that the use of FAP in the diets of finishing sheep reaped benefits on animal health and the development of rumen epithelium by improving antioxidant activity in plasma and stimulating the growth of papillae.

The aim of the present study was to describe age-related changes in anatomic, functional and microbial variables during the rumen development process, as affected by the feeding system (supplemental feeding v. grazing), in goats. Goats were slaughtered at seven time points that were selected to reflect the non-rumination (0, 7 and 14 d), transition (28 and 42 d) and rumination (56 and 70 d) phases of rumen development. Total volatile fatty acid (TVFA) concentration (P= 0·002), liquid-associated bacterial and archaeal copy numbers (P< 0·01) were greater for supplemental feeding v. grazing, while rumen pH (P< 0·001), acetate molar proportion (P= 0·003) and solid-associated microbial copy numbers (P< 0·05) were less. Rumen papillae length (P= 0·097) and extracellular (P= 0·093) and total (P= 0·073) protease activity potentials in supplemented goats tended to be greater than those in grazing goats. Furthermore, from 0 to 70 d, irrespective of the feeding system, rumen weight, rumen wall thickness, rumen papillae length and area, TVFA concentration, xylanase, carboxymethylcellulase activity potentials, and microbial copy numbers increased (P< 0·01) with age, while the greatest amylase and protease activity potentials occurred at 28 d. Most anatomic and functional variables evolved progressively from 14 to 42 d, while microbial colonisation was fastest from birth to 28 d. These outcomes suggest that the supplemental feeding system is more effective in promoting rumen development than the grazing system; in addition, for both the feeding systems, microbial colonisation in the rumen is achieved at 1 month, functional achievement at 2 months, and anatomic development after 2 months.

Two experiments were conducted to assess the effect of physical form of a starter feed with or without straw supplementation on growth performance of Holstein calves. In experiment 1, a total of 32 calves were randomly assigned at 7 d of age to texturized starter feed (containing rolled barley, corn, and oats) without straw, texturized starter feed with chopped straw, and pelleted starter feed with chopped straw. All calves were offered 4 L of pasteurized whole milk twice daily from 7 to 35 d of age, 2 L of milk twice daily from 36 to 42 d of age, and 2 L of milk from 43 to 49 d of age. Animals were weaned at 50 d of age, and the study finished when calves were 63 d old. In experiment 2, a total of 60 calves (8 d of age) were randomly assigned to texturized starter feed (containing whole corn) without straw, pelleted starter feed without straw, and pelleted starter feed with chopped straw. All calves were offered the same milk replacer (MR; 23% crude protein and 19.5 fat) at 11% dry matter concentration, 4 L/d of MR until 14 d of age, 6 L/d of MR from 14 to 37 d, 3 L/d of MR from 38 to 44 d, and 1.5 L/d of MR from 45 to 52 d of age. The experiment finished when calves were 58 d old (1 wk after weaning). Rumen liquid pH was measured after weaning. In both studies, calves were individually housed in pens on sawdust bedding and starter feed and chopped straw were offered free choice in separate buckets. In experiment 1, starter feed and straw intake and growth did not differ among treatments. However, calves receiving straw showed a greater rumen pH compared with those not receiving straw. In experiment 2, pelleted started feed supplemented with straw fostered an increase in solid feed intake (as percentage of body weight) compared with a pelleted or texturized starter feed without straw supplementation. However, calves that received the texturized starter feed containing whole corn had rumen pH similar to those fed a pelleted starter feed with straw. Feeding a texturized starter feed containing rolled barley, corn, and oats (with or without straw provision) was not able to maintain rumen pH or promote growth and intake compared with offering a pelleted starter feed with chopped straw. However, when whole corn was used in the texturized starter feed, rumen pH was equivalent to that obtained with a pelleted starter feed and straw supplementation., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

A slaughter experiment was conducted to determine the effects of alfalfa particle size on rumen morphology and performance of lambs. Twenty-four Balouchi lambs aged 21 days (9.1 ± 1.1 kg) were randomly fed control (diet without alfalfa hay; CON) and mixed rations containing 15% finely ground (FINE; 2 mm) and 15% coarsely chopped alfalfa hay (LONG; 3 to 4 cm). After a 63 days feeding period, nine animals (three per treatment) were slaughtered to obtain ruminal tissue samples for morphological analyses. Alfalfa particle size did not affect (p > 0.05) papillae density, height, width, epithelium depth and surface area. Coarse alfalfa decreased the stratum corneum and increased (p < 0.05) muscle depth compared with fine and control diets. Neither DNA content and nor RNA concentration of rumen tissue was affected by feeding different diets. Forage particle size did not affect the blood concentration of glucose, urea nitrogen (BUN), beta-hydroxybutyric acid (BHBA) and non-esterified fatty acids (NEFA). Dry matter intake and feed conversion ratio were higher for control diet; however, there were no significant differences between treatments for average daily gain. These data suggest that coarse alfalfa significantly reduces the stratum corneum and increases muscularity of rumen wall and tended to better feed conversion ratio., (Journal of Animal Physiology and Animal Nutrition © 2014 Blackwell Verlag GmbH.)

Salmonellosis is an insidious and potentially epidemic problem in pre-weaned dairy calves. Managing this disease, or any other diarrheal disease, is a financial burden to producers. Calf mortalities and medicinal treatments are overt costs of salmonellosis, while hidden costs include hampered weight gains and persistent intestinal colonization of the pathogen. In this study, we examined the anti-Salmonella effects of Saccharomyces cerevisiae fermentation products (SCFP) incorporated into both the milk replacer and the starter grain. In a blinded study, 2-8 day-old calves were fed SCFP (n=20 calves) or an SCFP-free Control (n=20 calves) for two weeks before and three weeks after experimental challenge with Salmonella enterica serotype Typhimurium. Following the challenge, calves were monitored for clinical signs and parameters associated with salmonellosis. Calves were then euthanized and examined for rumen development and intestinal Salmonella colonization. When compared to calves that received milk replacer and feed lacking SCFP, calves fed SCFP had fewer bouts of diarrhea and fever. Rumens from these calves were more developed, as measured by the length of papillae, which is consistent with the enhanced weight gain observed in this treatment group. Additionally, Salmonella intestinal colonization was reduced in SCFP-fed calves and Salmonella fecal shedding disappeared at an earlier stage in these calves. This study revealed that the combination of two proprietary S. cerevisiae fermentation products provide marked benefit for preventing the negative effects of salmonellosis in pre-weaned dairy calves, while also boosting productivity. The mechanism of action needs to be clarified, but it may be related to the observed decrease in colonization by the pathogen and increase in rumen development., (Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.)

Urea recycling, with urea originating from catabolism of amino acids and hepatic detoxification of ammonia, is particularly relevant for ruminant animals, in which microbial protein contributes substantially to the metabolizable protein supply. However, the quantitative contribution of urea recycling to protein anabolism in calves during the transition from preruminants (milk-fed calves) to ruminants [solid feed (SF)-fed calves] is unknown. The aim of this study was to quantify urea recycling in milk-fed calves when provided with low-protein SF. Forty-eight calves [164 ± 1.6 kg body weight (BW)] were assigned to 1 of 4 SF levels [0, 9, 18, and 27 g of dry matter (DM) SF · kg BW(-0.75) · d⁻¹] provided in addition to an identical amount of milk replacer. Urea recycling was quantified after a 24-h intravenous infusion of [¹⁵N₂]urea by analyzing urea isotopomers in 68-h fecal and urinary collections. Real-time qPCR was used to measure gene expression levels of bovine urea transporter B (bUTB) and aquaglyceroporin-3 and aquaglyceroporin-7 in rumen wall tissues. For every incremental gram of DM SF intake (g DM · kg(0.75)), nitrogen intake increased by 0.70 g, and nitrogen retention increased by 0.55 g (P < 0.01). Of this increase in nitrogen retention, 19% could be directly explained by urea recycling. Additionally, part of the observed increase in nitrogen retention could be explained by the extra protein provided by the SF and likely by a greater efficiency of postabsorptive use of nitrogen for gain. Ruminal bUTB abundance increased (P < 0.01) with SF provision. Aquaglyceroporin-3 expression increased (P < 0.01) with SF intake, but aquaglyceroporin-7 expression did not. We conclude that in addition to the increase in digested nitrogen, urea recycling contributes to the observed increase in nitrogen retention with increasing SF intake in milk-fed calves. Furthermore, ruminal bUTB and aquaglyceroporin-3 expression are upregulated with SF intake, which might be associated with urea recycling., (© 2014 American Society for Nutrition.)

Molecular mechanisms regulating rumen epithelial development remain largely unknown. To identify gene networks and regulatory factors controlling rumen development, Holstein bull calves (n=18) were fed milk replacer only (MRO) until 42 d of age. Three calves each were euthanized at 14 and 42 d of age for tissue collection to represent preweaning, and the remaining calves were provided diets of either milk replacer + orchard grass hay (MH; n=6) to initiate weaning without development of rumen papillae, or milk replacer + calf starter (MG; n=6) to initiate weaning and development of rumen papillae. At 56 and 70 d of age, 3 calves from the MH and MG groups were euthanized for collection of rumen epithelium. Total RNA and protein were extracted for microarray analysis and to validate detected changes in selected protein expression, respectively. As expected, calves fed MRO had no rumen papillae and development of papillae was greater in MG versus MH calves. Differentially expressed genes between the MRO diet at d 42 (preweaning) versus the MG or MH diets at d 56 (during weaning) were identified using permutation analysis of differential expression. Expression of 345 and 519 transcripts was uniquely responsive to MG and MH feeding, respectively. Ingenuity Pathway Analysis (Qiagen, Redwood City, CA) indicated that the top-ranked biological function affected by the MG diet was the cell cycle, and TFGB1, FBOX01, and PPARA were identified as key transcriptional regulators of genes responsive to the MG diet and associated with development of rumen papillae. Increased expressions of TGFB1 mRNA and protein in response to the MG diet were confirmed by subsequent analyses. The top-ranking biological function affected by the MH diet was energy production. Receptors for IGF-1 and insulin, ESRRA, and PPARD were identified by ingenuity pathway analysis as transcriptional regulators of genes responsive to the MH diet. Further analysis of TGFB1 and ESRRA mRNA expression in rumen epithelium obtained from a separate ontogenic study of Holstein calves (n=26) euthanized every 7d from birth to 42 d of age showed increases in transcript expression with advancing age, supporting their roles in mediating rumen epithelial development and function during weaning. Additional evaluation of gene expression in the rumen epithelium of adult cows ruminally infused with butyrate also suggested that observed changes in ESRRA mRNA expression in developing calf rumen may be mediated by increased butyrate concentration. Our results identify TGFB1 and ESRRA as likely transcriptional regulators of rumen epithelial development and energy metabolism, respectively, and provide targets for modulation of rumen development and function in the growing calf., (Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)