Technical note: A simple rumen collection device for calves: An adaptation of a manual rumen drenching system

A limited amount of research is available related to the rumen microbiota of calves, yet there has been a recent spike of interest in determining the diversity and development of calf rumen microbial populations. To study the microbial populations of a calf's rumen, a sample of the rumen fluid is needed. One way to take a rumen fluid sample from a calf is by fistulating the animal. This method requires surgery and can be very stressful on a young animal that is trying to adapt to a new environment and has a depressed immune system. Another method that can be used instead of fistulation surgery is a rumen pump. This method requires a tube to be inserted into the rumen through the calf's esophagus. Once inside the rumen, fluid can be pumped out and collected in a few minutes. This method is quick, inexpensive, and does not cause significant stress on the animal. This technical note presents the materials and methodology used to convert a drenching system into a rumen pump and its respective utilization in 2 experiments using dairy bull calves.     

Technical note: Infusion,sampling, and vacuum-assisted collection devices for use in ruminally cannulated calves

Calves can be ruminally cannulated at young ages, but equipment size limitations preclude use of an infusion and sampling device in these small animals. Likewise, a procedure to easily evacuate rumen contents in young calves has not been described. Overcoming these technical complications related to assessment of ruminal passage kinetics, nutrient digestion, and volatile fatty acid absorption would aid in future studies advancing our knowledge of dairy calf nutrition. The first objective was to design and fabricate 2 devices (one device for infusion and sampling, and another for vacuum-assisted collection) suitable for use in young ruminally cannulated dairy calves. The second objective was to test the utility of these tools when performing procedures commonly used in ruminant nutrition research. A single weaned 62-d-old ruminally cannulated calf was used to evaluate the ability to infuse a solution of LiCoEDTA and sample rumen contents through the cannula cap over a period of 2 h to assess the rumen liquid passage rate (procedure 1). The device was capable of infusing the LiCoEDTA and sampling the rumen fluid, as evidenced by the presence of elevated Co concentrations in the sampled rumen fluid. Using the fluid samples obtained, liquid passage rate within the calf was estimated to be 40.2% of ruminal fluid/h. The second procedure tested the vacuum-assisted collection device and consisted of evacuating and weighing the rumen contents, which is considered a key preparatory step in washed reticulorumen technique experiments that aim to measure nutrient absorption. In agreement with existing literature, evacuated rumen contents represented approximately 4% of the calf's body weight. In conclusion, custom-built devices for infusion, sampling, and vacuum-assisted collection were efficacious when tested in a 62-d-old ruminally cannulated calf fed a diet of 100% texturized starter (18% crude protein, as-fed). Fellow scientists may employ and further modify these techniques to suit their needs when assessing passage kinetics, nutrient digestion, and volatile fatty acid absorption in calves.     

Influence of weaning method on health status and rumen development in dairy calves

In the artificial rearing of dairy calves, the same feeding plan is applied to all animals during the milk-feeding period, with individual differences attributable to development or health status rarely considered. The aim of this study was 1) to analyze whether the parameters of feeding behavior automatically recorded by a feeding computer and weight gain are suitable for predicting the health status and rumen development of male dairy calves, and 2) to compare a conventional weaning method (end of milk provision at 12 wk of age, n = 23 calves) with a concentrate-dependent weaning method (with reduction in the milk amount depending on the consumption of concentrate, n = 24). The health status of each animal was evaluated daily by a scoring list (health score), and body temperature was measured automatically during each milk intake. In addition, the number of veterinary treatments per calf was recorded. Rumen development was assessed by measuring rumen papillae in 8 rumen areas after slaughter (n = 24, half of each treatment group). During the milk-feeding period, body temperature was elevated (≥39.5°C) on 40.8 and 43.2% of all days for calves on the concentrate-dependent weaning method and the conventional weaning method, respectively. Hay and concentrate intake (but not milk intake) and weight gain were clearly affected by health status. In addition, health score and the probability of being treated by a veterinarian were significantly related to decreases in concentrate consumption. During the milk-feeding period, increased body temperature, an increased number of veterinary treatments, and decreases in milk consumption were all associated with reduced weight gain. Calves on the concentrate-dependent weaning method were weaned at an average age of 76 d, which was significantly shorter than the age at the end of milk provision for conventionally fed calves (84 d). Weight gain and health status did not differ between treatment groups. Weight gain was positively associated with papillae length. A treatment effect on rumen development could not be found. We conclude that the concentrate-dependent weaning method allows a faster physiological development without any negative impact on rumen development, weight gain, or health status; we therefore recommend its use in practice.     

Effect of various levels of forage and form of diet on rumen development and growth in calves

The effect of form of starter grain (coarse vs. ground) and inclusion of various levels of hay on body weight gain and rumen development was evaluated. Two experiments were conducted to determine the effect of form of diet and forage inclusion on intake, growth, feed efficiency, and weaning age in dairy calves. Diets consisted of commercial coarse starter (C), ground starter (G), coarse starter with 7.5% bromegrass hay of consistent particle size (8 to 19 mm) (H1), and coarse starter with 15% hay (H2). In experiment 1, intake was held constant across treatments until weaning, when feed was offered ad libitum. Calves receiving H1 and H2 were heavier and had greater body weight gain and greater feed efficiency than calves receiving C. There were no differences in intake. Total volatile fatty acid concentrations were higher, and the proportion of acetate was lower for calves fed G vs. C. In experiment 2, calves (n = 56) were offered diets on an ad libitum basis and weaned according to intake. There were no differences in body weight gain, average daily gain, feed efficiency, and age at weaning with respect to treatment. Starter and total dry matter intake tended to be greater in calves fed H1 and H2 vs. C. The addition of controlled particle size hay to diets of young calves appears to favorably alter rumen environment, resulting in increased intake and improved feed efficiency. Forage of a consistent particle size can be successfully utilized in starter rations of young calves.     

Effects of supplemental yeast (Saccharomyces cerevisiae) culture on rumen development, growth characteristics, and blood parameters in neonatal dairy calves

Yeast (Saccharomyces cerevisiae) culture was added to a texturized calf starter at 0 (control), 1, or 2% of dry matter to determine effects on intake, growth, blood parameters, and rumen development. Seventy-five Holstein calves (38 male; 37 female) were started on the experiment at 2 +/- 1 d of age and were studied for 42 d. Starter intake was measured, and fecal scoring was conducted daily. Growth and blood parameter measurements were recorded at weekly intervals. A subset of 6 male calves (2 per treatment) was euthanized at 5 wk of age, and rumen tissue was sampled for rumen epithelial growth measurements. An additional 6 male calves were euthanized at 6 wk of age for rumen epithelial growth measurements. Inclusion of yeast culture at 2% of the starter ration significantly increased starter and total dry matter intake, average daily gain, and daily hip width change when compared with the control treatment. Average daily gain was improved by 15.6% for the 2% yeast treatment. Daily change in hip height was also significantly greater for calves receiving 2% supplemental yeast culture than for calves receiving 1%. No significant treatment differences were observed for any other variables. These data suggest that the addition of yeast culture in a dairy calf starter at 2% enhances dry matter intake and growth and slightly improves rumen development in dairy calves.     

Preweaning to postweaning rumen papillae structural growth,ruminal fermentation characteristics,and acute-phase proteins in calves

This study evaluated pre- to postweaning ruminal structural development, fermentation characteristics, and acute-phase protein levels in calves with a high milk replacer (MR) feeding rate prior to weaning. Six ruminally cannulated Holstein bull calves were fed MR (150 g/L) at 15% of body weight (BW) in 2 equal volumes daily. Volumes were adjusted weekly based on BW. Calves were weaned using a 1-step weaning method, with MR decreased by 50% at the end of wk 5 and full weaning at the end of wk 6. Calf starter, chopped straw, and water were offered ad libitum. Intake was recorded daily, and BW was recorded weekly. From wk 5 to 12, ruminal pH was continuously measured using a ruminal pH bolus. Ruminal fluid was collected weekly from wk 5 to 12 for measurement of short-chain fatty acid concentrations and quantification of total bacteria and protozoa. Rumen papillae were obtained at wk 5, 6, 7, 8, and 12 for histological analysis. Serum amyloid A and lipopolysaccharide-binding protein were measured weekly. Data were analyzed using GLIMMIX procedure of SAS (SAS Institute Inc., Cary, NC), with week as a fixed effect and calf as a random effect. During the weaning step-down, starter intake was 3-fold higher and continued to increase until wk 12. Body weight increased from birth to wk 12; however, BW did not change during wk 6, 7, and 8, possibly due to low metabolizable energy intake caused by the weaning strategy. Preweaning ruminal pH was below 5.8 for approximately 936.3 ± 125.99 min/d, implying ruminal acidosis. Furthermore, ruminal pH below 5.8 reached a peak at wk 8 with 1,203.9 ± 227.65 min/d below pH 5.8 and slowly decreased to 388.1 ± 189.82 min/d below pH 5.8 at wk 12. Papillae surface area, length, and width increased during wk 12 compared with wk 5. Corneum thickness increased by week, whereas spinosum/basale thickness only increased during wk 8 compared with wk 5. The acute-phase protein concentration was highest at wk 1 and then decreased and remained constant until wk 12. In conclusion, even before step-down weaning, calves experienced ruminal acidosis despite low starter intake. Further, the observed prolonged ruminal pH depression suggests that dietary rumen adaptation after weaning can take several weeks in calves with a high MR feeding rate preweaning. The prolonged depressed ruminal pH did not affect acute-phase proteins and this finding, along with the other results, suggests that rumen epithelium barrier integrity is not compromised during weaning.     

Starch source evaluation in calf starter: II. Ruminal parameters, rumen development, nutrient digestibilities, and nitrogen utilization in Holstein calves

Ruminal parameters, rumen development, nutrient digestibilities, and N utilization were estimated in Holstein calves fed starch from different sources. Ground corn, ground barley, ground wheat, and crimped oats were used to formulate 4 isostarch (25% of starter dry matter) pelleted diets. These diets were randomly allocated to calves (16 calves per treatment, 8 female and 8 male) and fed ad libitum along with mixed grass hay throughout the experiment. Ruminal contents and blood were sampled at d 35, 50, and 70 of age to estimate ruminal parameters and plasma β-hydroxybutyrate, respectively. At d 70, twenty-four male calves (6/treatment) were randomly selected, euthanized, and forestomach weight, papillae length (PL), papillae width (PW), rumen wall thickness (RWT), and papillae concentration were measured. At d 63, twenty-four female calves (6/treatment) were randomly selected and moved to metabolism stalls to estimate total tract apparent nutrient digestibilities and N utilization. Female calves were given 2 wk for adaptation to experimental facilities and then total collections of feces and urine were made from d 77 to 84 of age. Ruminal pH at d 35 of age was higher in calves fed corn and oat diets than in those fed barley and wheat diets. Ruminal pH at d 50 and 70 of age was the lowest in calves on barley diets followed by those on oat and wheat diets and then by those on the corn diet. Ruminal total volatile fatty acid concentrations at d 35 of age were greatest in calves fed corn or wheat diets followed by those fed barley and oat diets. Calves on corn and wheat diets maintained greater ruminal volatile fatty acids concentrations at d 50 and 70 of age. Ruminal ammonia, acetate, propionate, butyrate, and blood β-hydroxybutyrate concentrations were also greater in calves on the corn and wheat diets. Full and empty weights of forestomach, PL, PW, RWT, and papillae concentrations were greater in calves on corn and wheat diets. Daily average intake of nutrients (dry matter, crude protein, neutral detergent fiber, starch, Ca, and P) was greater in calves fed corn and wheat diets than in those fed barley and oat diets. Starch source did not influence the total tract apparent digestibilities of nutrients in calves. Daily N retention (g/d) was greatest on the corn diet followed by the wheat diet and then the barley and oat diets. In conclusion, calves on a corn diet have greater ruminal capacity to accommodate feed bulk. More physically and metabolically functional rumens in calves on corn and wheat diets probably resulted in greater feed consumption and N retention.     

Short communication: Effect of calf starter on rumen pH of Holstein dairy calves at weaning

The objective of this study was to evaluate the effect of feeding calf starter on rumen pH of dairy calves during weaning transition. Twenty Holstein bull calves were paired into 10 blocks by starting date of the study and body weight, and fed either milk replacer and hay (MR) or MR, hay, and a commercial texturized calf starter (MR+S) in a randomized complete block design. All calves were fed 750 g/d of milk replacer as the basal diet. Calves on MR+S treatment were also fed a calf starter ad libitum to maintain similar energy intake between calves within blocks, and MR calves were fed additional milk replacer that was equivalent to energy from calf starter intake. When MR+S calves consumed a calf starter at 680 g/d for 3 consecutive d, rumen pH of a MR+S calf and his MR counterpart was measured continuously for 3 d using a small ruminant rumen pH measurement system. Treatment did not affect minimum pH, mean pH, maximum pH, standard deviation of mean pH, and duration or area under pH 5.8, indicating that calf starter consumption did not appear to affect rumen pH. However, hay intake was negatively correlated to area under pH 5.8, with a breakpoint at 0.080 kg/d intake, suggesting hay intake might play an important role in mitigating ruminal acidosis in dairy calves during weaning transition.     

Technical note: The development of a methodology for ruminal and colon tissue biopsying of young Holstein dairy calves

The objectives of this study were to develop a methodology for biopsying the rumen and colon of young dairy calves and to collect suitable quality tissue samples for microscopic and gene expression analysis. Six Holstein dairy bull calves (45.0 ± 1.5 kg birth weight) were ruminally cannulated during the second week of life and weaned at the end of wk 6. Ruminal and colon tissue samples were collected at the end of wk 5, 6, 7, 8, and 12. Calves were not sedated but were restrained in a chute for sampling. The endoscope (100 cm length, 9.8 mm diameter) was introduced through the rumen cannula to harvest ruminal tissue. Endoscopic biopsies of the rumen with endoscopic biopsy forceps were unsuccessful 85% of the time because they were unable to shear the ruminal tissue. Thereafter, an Allis clamp was used to retrieve the blind sac through the rumen cannula to perform direct tissue biopsying with surgical scissors. To biopsy the colon, the lubricated distal tip of an endoscope was slowly inserted into the calf's anus. A total of 6 colon tissue samples (12.6 ± 0.74 mg) were collected per calf per time point from the distal colon 30 to 40 cm from the calf's anus using endoscopic biopsy forceps, which were inserted through the instrument channel. A new forcep was used between sites and calves. Between calves, the outside of the endoscope was washed with 4% chlorohexidine and rinsed with water and the instrument channel was washed with distilled water and 70% ethanol. Colon and ruminal samples were processed for histological measurements, and RNA was isolated and sequenced. High-quality RNA (RNA integrity number 8.8 ± 0.08) was collected from samples, and light and electron microscopy was performed on samples. In conclusion, endoscopic biopsying can be used for tissue harvest in the colon of young calves. However, it was found that collecting ruminal tissue by retracting the rumen from the cannula and taking samples with surgical scissors was more successful than an endoscopic biopsy. This method allows for tissue collection of the same animal throughout time, which can help the research community investigate the effect of weaning regimens, feed rations, and age on the structure and function of the gastrointestinal tract.     

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

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

Evaluation of a system for monitoring rumination in heifers and calves

The Hi-Tag electronic rumination-monitoring system (SCR Engineers Ltd., Netanya, Israel), based on capturing audio recordings, provides a reasonable measure of rumination time in dairy cows, but has not been validated for milk-fed or weaned heifers. The objective of this study was to validate the Hi-Tag rumination-monitoring system in heifers and calves and to assess whether suckling from a teat interfered with recording from this system. Assessments of 2 independent observers were highly correlated (r = 0.99, n = 20), indicating that direct visual observations provide a useful standard. Measures from the Hi-Tag system were validated by comparing values with those from a single human observer, using observations from three 2-h intervals from 35 Holstein calves and heifers aged 4, 6, and 9 wk and 3, 6, and 9 mo, respectively. In 9-mo-old heifers, rumination times obtained from the electronic system were highly correlated with visual observations (r = 0.88, R² = 0.77, n = 15), and the mean difference was minor (−4 ± 8 min/2-h interval). The accuracy of data from the automated system decreased when used on heifers less than 9 mo old. Suckling did not interfere with the electronic system (r = −0.1, n = 18). These results indicate that the Hi-Tag system is an accurate tool for monitoring rumination behavior in Holstein Friesian heifers from the age of 9 mo.     

Longitudinal assessment revealed the shifts in rumen and colon mucosal-attached microbiota of dairy calves during weaning transition

The objectives of this study were to investigate the shifts in rumen and colon mucosa-associated microbiota in dairy calves fed a high milk replacer feeding rate before and after weaning and to determine whether such shifts are associated with tissue physiological measures. Longitudinal biopsy was performed to collect rumen and colon mucosal tissues of 4 ruminally cannulated Holstein dairy bull calves (weaned at 6 wk of age) at the end of wk 5 (before weaning), 7 (weaning adaptation) and 12 (after weaning), and were used to assess mucosa-associated microbiota and their changes using amplicon sequencing. Both rumen and colon mucosa-associated bacterial communities shifted during the weaning process, as evidenced by their clear separation among 3 different weaning periods and increased α diversity (Shannon and Chao1 indices) during weaning transition. Among the 3 dominant bacterial phyla identified (relative abundance >1.0%), the relative abundance of Proteobacteria and Bacteroidetes decreased in the rumen mucosa, whereas the relative abundance of Firmicutes increased in both rumen and colon mucosa during weaning transition. In the rumen mucosa, Campylobacter (0.6–22.1%) gradually became prevalent during weaning transition, whereas Succinivibrio (6.2–10.3%) and Prevotella 1 (4.7–10.5%) were dominant regardless of weaning transition. In the colon mucosa, Bacteroides (12.8–25.4%) was dominant during weaning transition, although its relative abundance decreased after weaning. In the meantime, relative abundance of uncultured Lachnospiraceae increased from 2.2% to 25.7% during this period. In addition, genera Pyramidobacter (in the rumen mucosa) and Lachnoclostridium (in the colon mucosa) were positively correlated with rumen papilla surface area and colon mucosal thickness, respectively. Moreover, genera Ruminococcaceae UCG-005 and Sharpea in the rumen mucosa were positively correlated with the molar proportion of propionate and butyrate, respectively. Overall, our findings revealed that rumen and colon mucosa-associated bacterial communities altered in response to the weaning transition, and some bacterial taxa in these communities may have positive effects on rumen and colon mucosa development during this period.     

Technical note: Evaluation of a triaxial accelerometer for monitoring selected behaviors in dairy calves

The objectives of this study were (1) to develop an algorithm for the acceleration sensor of the Smartbow Eartag (Smartbow GmbH, Weibern, Austria) to distinguish between postures (lying and standing or locomotion) and to detect 6 kinds of activities (milk intake, water intake, solid feed intake, ruminating, licking or sucking without milk intake, and other activities) in dairy calves and (2) to evaluate this sensor for identifying these behaviors in dairy calves compared with observations from video. Accelerometers were applied to the left ears of 15 preweaned Holstein dairy calves. Calves were kept in a group pen and received milk replacer from an automatic calf feeder. Based on 38 h of acceleration data and video observation, an algorithm was established to detect the predefined behaviors. Using cross-validation, video recordings were used to analyze whether a behavior was detected correctly by the developed algorithm. For posture, sensitivity (94.4%), specificity (94.3%), precision (95.8%), and accuracy (94.3%) were high. Cohen's kappa was calculated as 0.88. For the 6 defined activities, overall (i.e., aggregated for all activities) accuracy was 70.8% and kappa was calculated as 0.58. Some activities (e.g., ruminating, feed intake, other activities) were identified better than others. In conclusion, the developed algorithm based on the acceleration data of the Smartbow Eartag was successful in detecting lying behavior, rumination, feed intake, and other activities in calves, but further development of the underlying algorithm will be necessary to produce reliable results for milk and water intake.     

Effect of roughage source and roughage to concentrate ratio on animal performance and rumen development in veal calves

Sixty-four male Holstein-Friesian × Dutch Friesian veal calves (46 ± 3.0 kg) were used to evaluate the effect of the inclusion of different levels and sources of dietary roughage on animal performance and rumen development. Treatments consisted of 1) C100 = concentrate only; 2) C70-S30 = concentrate (70%) with straw (30%), 3) C70-G30 = concentrate (70%) with dried grass (30%), 4) C70-G15-S15 = concentrate (70%) with dried grass (15%) and straw (15%), 5) C70-CS30 = concentrate (70%) with corn silage (30%), 6) C40-CS60 = concentrate (40%) with corn silage (60%), 7) C70-CS30-AL = concentrate (70%) with corn silage (30%) ad libitum, 8) C70-G15-S15-AL = concentrate (70%) with dried grass (15%) and straw (15%) ad libitum. All dietary treatments were provided in addition to a commercial milk replacer. Concentrate was provided as pellets and roughage was chopped. The dietary treatments 1 to 6 were supplied restrictedly to a maximum of 750 g of dry matter (DM) per day, whereas treatments 7 and 8 were offered ad libitum in combination with a reduced amount of milk replacer. Calves were euthanized after 10 wk. Straw supplementation (C70-S30 vs. C70-G30 and C70-CS30) reduced DM intake, and ad libitum supply of concentrate and roughage increased DM intake. Roughage addition did not affect growth performance. Rumen fermentation was characterized by low pH and high total volatile fatty acids and reducing sugar concentrations. Calves fed ad libitum showed lower ruminal lactate concentrations than calves fed restrictedly. Ammonia concentrations were highest in calves fed C-100 and lowest in calves fed ad libitum. The recovery of CoEDTA (added to milk replacer) varied between 20.5 and 34.9%, indicating that significant amounts of milk entered the rumen. Roughage addition decreased the incidence of plaque formation (rumen mucosa containing focal or multifocal patches with coalescing and adhering papillae covered by a sticky mass of feed, hair and cell debris) and the incidence of calves with poorly developed rumen mucosa. However, morphometric parameters of the rumen wall were hardly influenced by the type and level of roughage. Ruminal polysaccharide-degrading enzyme activities reflected the adaptation of the microorganisms to the dietary concentrate and roughage source. Results indicated that in veal calves, the addition of roughage to concentrate diets did not affect growth performance and positively influenced the macroscopic appearance of the rumen wall.     

Technical note: validation of methodology for characterization of feeding behavior in dairy calves

Time sampling techniques are useful in collecting feeding behavior data because they minimize the time required for observation. Instantaneous recording is often used in the collection of feeding behavior data for dairy calves; however, the recording intervals used vary widely and it is unclear what minimum interval is necessary to yield accurate data. The objective of this study was to validate data obtained using instantaneous recording of feeding behavior of dairy calves across a range of time intervals with data obtained from continuous recording. Ten Holstein bull calves were observed continuously using time-lapse video for 3 d during the milk-feeding period while they were fed milk ad libitum and for 3 d post-milk-weaning while they consumed solid feed. Feeding behavior data obtained from continuous recording were compared with data obtained from instantaneous recording at intervals ranging from 15 s to 10 min. As expected, the strength of linear association between behavior measures obtained from continuous recording and instantaneous recording decreased with increasing recording interval. The relationship varied between feeding behavior measures; feeding time was represented well (R² >0.76) by instantaneous recording at up to 5-min intervals, but a strong linear association of meal frequency and meal time (R² >0.8) required intervals no greater than 1 min and 30 s, respectively. The relationship between feeding behavior measures obtained from continuous recording and recording at different intervals was similar in both periods; however, sensitivity of time sampling data across recording intervals was greater during the milk-feeding period. Sensitivity was low in both periods (<0.7 with scanning intervals as short as 1 min), indicating that instantaneous recording may not represent within-meal feeding behavior well. Instantaneous recording can provide accurate calf feeding behavior data if the recording interval is sufficiently short.     

Technical note: An improved tool to insert lines for abomasal infusion in rumen cannulated cattle

An improved tool was tested that facilitates the insertion of abomasal infusion lines in ruminally cannulated cattle for postruminal infusion studies. The insertion device was made from a 68 × 25 mm section of 20-mm i.d. stainless steel device with a weight of 125 g. The general procedure to place the tool is to pass the insertion device with the infusion line through the rumen cannula, through the sulcus omasi (reticulo-omasal orifice), and finally into the abomasum. Once inside the omasum, the insertion device is placed inside the abomasum by gently shifting the infusion line from the flexible polyvinyl chloride tubing fitted to the insertion device. The insertion device places itself by self-moving down through the abomasal orifice into the abomasum by means of its own weight. After device placement, its position can be verified by palpating through the wall of the rumen. This way, an infusion line can easily be inserted into the abomasum by any person without introducing the hand into the omasum, avoiding possible injuries to the animal and tool being expulsed of its desired position.     

Interactions of alfalfa hay and sodium propionate on dairy calf performance and rumen development

The objective of this experiment was to investigate the effects of different levels of alfalfa hay (AH) and sodium propionate (Pro) added to starter diets of Holstein calves on growth performance, rumen fermentation characteristics, and rumen development. Forty-two male Holstein calves (40 ± 2 kg of birth weight) were used in a complete randomized design with a 3 × 2 factorial arrangement of treatments. Dietary treatments were as follows: (1) control = concentrate only; (2) Pro = concentrate with 5% sodium propionate [dry matter (DM) basis]; (3) 5% AH = concentrate + 5% alfalfa hay (DM basis); (4) 5% AH + Pro = concentrate + 5% alfalfa hay + 5% sodium propionate (DM basis); (5) 10% AH = concentrate + 10% alfalfa hay (DM basis); and (6) 10% AH + Pro = concentrate + 10% alfalfa hay + 5% sodium propionate (DM basis). All calves were housed in individual pens bedded with sawdust until 10 wk of age. They were given ad libitum access to water and starter throughout the experiment and were fed 2 L of milk twice daily. Dry matter intake was recorded daily and body weight weekly. Calves from the control, 10% AH, and 10% AH + Pro treatments were euthanized after wk 10, and rumen wall samples were collected. Feeding of forage was found to increase overall dry matter intake, average daily gain, and final weight; supplementing sodium propionate had no effect on these parameters. Calves consuming forage had lower feed efficiency than those on the Pro diet. Rumen fluid in calves consuming forage had higher pH and greater concentrations of total volatile fatty acids and molar acetate. Morphometric parameters of the rumen wall substantiated the effect of AH supplementation, as plaque formation decreased macroscopically. Overall, the interaction between forage and sodium propionate did not affect calf performance parameters measured at the end of the experiment. Furthermore, inclusion of AH in starter diets positively enhanced the growth performance of male Holstein calves and influenced both the macroscopic and microscopic appearances of the rumen wall. These benefits, however, were small when only sodium propionate was offered.     

Effects of a mixture of phytobiotic-rich herbal extracts on growth performance,blood metabolites,rumen fermentation,and bacterial population of dairy calves

Forty-eight newborn Holstein dairy calves [40 ± 3.4 (SD) of kg of body weight (BW); 24 females and 24 males] were used in a completely randomized design to investigate the effects of a mixture of phytobiotic-rich herbal extract (Immunofin, IMPE) incorporated into milk on performance, ruminal fermentation, bacterial population, and serum biochemical metabolites during the preweaning period. Calves had free access to calf starter and clean water from d 6 until weaning. The treatments were the control (CON; without additive) and IMPE at 4, 8, and 12 mL/d. The treatments had no significant effect on total dry matter intake, weight gain, and BW at weaning. The incidence of diarrhea was lower in calves fed 8 mL of IMPE/d compared with CON. At weaning, body measurements (except for front leg circumference) were not affected by IMPE treatment. Relative to the CON group, front leg circumference was significantly decreased by IMPE supplementation. Serum IgG concentration was not significantly increased by IMPE supplementation compared with the CON group. Triglyceride concentration decreased in calves receiving 4, 8, and 12 mL/d of IMPE compared with the CON groups. In contrast to the CON group, serum albumin and total serum protein concentrations increased with IMPE supplementation. Calves receiving 4 mL/d of IMPE had a greater abundance of total bacteria, Ruminococcus albus, Ruminococcus flavefaciens, and Fibrobacter succinogenes compared with the other treatments. Molar proportions of acetate increased in calves fed IMPE (at 12 mL/d) compared with calves fed CON. Ruminal N-NH₃ concentrations decreased linearly with the increase in IMPE supplementation. The results of the present study suggest that the addition of IMPE to milk may improve some health and immunity conditions, blood metabolite concentrations, and increase the abundance of some cellulolytic bacteria in the rumen of Holstein dairy calves. The use of IMPE may be an alternative to feeding antibiotics at subtherapeutic concentrations to improve calf health and immunity status.     

Effects of forage provision to young calves on rumen fermentation and development of the gastrointestinal tract

Fifteen Holstein male calves were randomly assigned to 1 of 3 dietary treatments according to age and body weight (BW) to determine the effects of feeding different forages sources on rumen fermentation and gastrointestinal tract (GIT) development. Treatments consisted of a starter (20% crude protein, 21% neutral detergent fiber) fed alone (CON) or supplemented with alfalfa (AH) or with oat hay (OH). All calves received 2 L of milk replacer (MR) at 12.5% dry matter twice daily until 49 d of age. Calves received 2 L of the same MR from 50 to 56 d of age and were weaned at 57 d of age. Individual starter, forage, and MR intakes were recorded daily and BW was recorded weekly. A rumen sample was taken weekly to determine rumen pH and volatile fatty acid concentrations. Three weeks after weaning, animals were harvested and each anatomical part of the GIT was separated and weighed with and without contents. Rumen pH was lower in CON than in OH and AH calves. Furthermore, acetate proportion in the rumen liquid tended to be greater in AH than in CON and OH treatments. Total GIT weight, expressed as a percentage of BW, tended to be greater in AH compared with the other 2 treatments. Rumen tissue tended to weigh more in CON than in OH animals. Animals with access to forage tended to have a greater expression of monocarboxylate transporter 1 than CON calves. In conclusion, calves supplemented with oat hay have a better rumen environment than calves offered no forage and do not have an increased gut fill.     

Rumen microbiota and its relation to fermentation in lactose-fed calves

In our previous studies, we revealed the effect of lactose inclusion in calf starters on the growth performance and gut development of calves. We conducted the present study as a follow-up study to identify the shift in rumen microbiota and its relation to rumen fermentation when calves are fed a lactose-containing starter. Thirty Holstein bull calves were divided into 2 calf starter treatment groups: texturized calf starter (i.e., control; n = 15) or calf starter in which starch was replaced with lactose at 10% (i.e., LAC10; n = 15) on a dry matter basis. All calves were fed their respective treatment calf starter ad libitum from d 7, and kleingrass hay from d 35. Rumen digesta were collected on d 80 (i.e., 3 wk after weaning) and used to analyze rumen microbiota and fermentation products. There was no apparent effect of lactose feeding on the α-diversity and overall composition of rumen microbiota. Amplicon sequencing and real-time PCR quantification of the 16S rRNA gene confirmed that the abundance of butyrate-producing bacteria (i.e., Butyrivibrio group and Megasphaera elsdenii) did not differ between the control and LAC10 groups. Conversely, the relative abundance of Mitsuokella spp., which produce lactate, succinate, and acetate, was significantly higher in the rumen of calves that were fed lactose, whereas the lactate concentration did not differ between the control and LAC10 groups. These findings suggest that the lactate production can be elevated by an increase of Mitsuokella spp. and then converted into butyrate, not propionate, since the proportion of propionate was lower in lactose-fed calves. In addition, we observed a higher abundance of Coriobacteriaceae and Pseudoramibacter-Eubacterium in the LAC10 group. Both these bacterial taxa include acetate-producing bacteria, and a positive correlation between the acetate-to-propionate ratio and the abundance of Pseudoramibacter-Eubacterium was observed. Therefore, the higher abundance of Coriobacteriaceae, Mitsuokella spp., and Pseudoramibacter-Eubacterium in the rumen of lactose-fed calves partially explains the increase in the proportion of rumen acetate that was observed in our previous study.