Technical note: technique for dissection and analysis of the rumen in young calves

This paper discusses a technique used to evaluate rumen development in young calves, including removal, dissection, and analysis of tissue. The method allowed for examination of the different sacs of the rumen (dorsal, ventral, cranial, and caudal) using scanning electron microscopy to measure papillae denseness and histology slides to measure papillae length and width. Computer software was used to produce accurate measurements of papillae. The rumens of young calves were dissected, and samples were taken from the cranial, caudal, ventral, and dorsal sections. Calves were part of a nutrition research study, and dietary treatments did have an effect on development measurements such as length, width, and papillae denseness.     

Technical note: development of a tool to insert abomasal infusion lines into dairy cows

A tool was developed to aid in ruminal insertion of abomasal infusion lines into dairy cows. The tool consisted of 2 pieces cut from polyvinyl chloride pipe. The first piece of pipe, the insertion tool, contained a groove that held the flexible plastic flange that is on the end of the infusion line. The insertion tool containing the flange was inserted into the ruminal cannula, through the sulcus omasi, and into the abomasum. The second piece of pipe, the delivery tool, was threaded through the insertion tool, and it was used to dislodge the flange from the insertion tool and into the abomasum.     

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.     

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: 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.     

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.     

Effects of active dried Saccharomyces cerevisiae on ruminal fermentation and bacterial community during the short-term ruminal acidosis challenge model in Holstein calves

We investigated the effects of active dried Saccharomyces cerevisiae (ADSC) on ruminal pH, fermentation, and the fluid bacterial community during the short-term ruminal acidosis challenge. Five rumen-fistulated male Holstein calves (147.0 ± 5.8 kg of body weight; 3.6 ± 0.2 mo of age) were used in a crossover design, and 0 g (control group, n = 5) or 2 g (SC group, n = 5) of ADSC (1 × 10¹⁰ cfu/g) was administered twice daily for 21 consecutive days. Calves were fed a high-forage diet during the first 15 d (d –14 to d 0; prechallenge), a high-grain diet for 2 d (d 1 and 2; ruminal acidosis challenge), and a high-forage diet for 4 d (d 3 to 6; postchallenge). Ruminal pH was measured continuously. Rumen fluid samples were collected once daily (0800 h) on d 0, 3, 4, and 6 and twice daily (0800 and 1100 h) on d 1 and 2. Bacterial DNA was extracted from fluid samples collected on d 0 and 3. The 24-h and 1-h mean ruminal pH was significantly depressed during the ruminal acidosis challenge in each group, although the changes were more severe in the SC group, consistent with a significant increase in lactic acid on d 2 (1100 h) compared with d 0 and a significantly higher proportion of butyric acid on d 2 (1100 h) compared with the control group. Feeding a high-grain diet caused a decrease in bacterial diversity due to high acidity in both groups. The relative abundances of the genus Bifidobacterium and operational taxonomic unit (OTU) 3 (Bifidobacterium species) increased significantly in both groups but were higher in the SC group. Correlation analyses indicated that OTU3 (Bifidobacterium species) were positively correlated with lactic acid concentration and that OTU1 (Prevotella species) and OTU5 (Succinivibrio species) were correlated with the proportion of butyric acid. These results suggest that ADSC supplementation induced the intense decreases in ruminal pH by increased butyric and lactic acid production through a high-grain diet fermentation by rumen fluid bacterial species during the short-term ruminal acidosis challenge in Holstein calves after weaning.     

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.     

Technical note: Evaluating nuclear magnetic resonance spectroscopy for determining body composition in Holstein dairy calves using deuterium oxide dilution methods

Deuterium oxide (D₂O) dilution methods have been used to assess body composition in live animals. Estimated body water content can be used to predict body fat and protein, and thus, the amount of energy reserves. It is an alternative method to direct chemical analysis and considered a noninvasive technique that is economical and repeatable. Deuterium oxide use is considered easy, safe, and accurate; however, the traditional methods of analyzing D₂O are expensive, tedious, and time consuming. The objective of this study was to evaluate the potential for using nuclear magnetic resonance spectroscopy (NMR) to determine body composition in Holstein dairy heifers. Nuclear magnetic resonance is less expensive and requires minutes to calculate the percentage of D₂O in the blood. This study used 24 newborn dairy heifer calves blocked by birth and randomly assigned to 1 of 3 treatments: (1) 446 g dry matter (DM) of a conventional milk replacer (MR) [CON; 20% crude protein (CP), 20% fat], (2) 669 g DM of a moderately high protein MR (MOD; 26% CP, 18% fat), or (3) 892 g DM of a moderately high protein MR (aggressive, AGG; 26% CP, 18% fat). All calves had free-choice access to starter and water. Both MR and starter were medicated with decoquinate. During weaning (d 43 to 49), the morning MR feeding ceased. On d 50, all MR feedings ended but starter and water intakes were continuously recorded until d 56. When calves were 50 d of age, a baseline blood sample was taken followed by injection of 300 mg of D₂O/kg of body weight in sterile physiological saline (0.9%). The syringes containing the D₂O in physiological saline were weighed before and after administration to record the actual dose of D₂O injected gravimetrically. After injection, the D₂O was allowed to equilibrate with body water for 1 h. Six blood samples were taken over 6 d (1/d) at 1630 h to estimate the dilution of the tracer. The plasma was aspirated and stored at ?20°C until further D₂O analysis. This new method was validated using 4 calf plasma samples that were sent to an outside laboratory for measurement using an independent validation method. We detected no differences in total body water, protein, fat, or mineral content in calves fed CON, MOD, or AGG; however, results demonstrated that the D₂O dilution technique and analysis by NMR is an appropriate and easy method to estimate water, protein, ash, and fat in young heifers.     

Evaluation of an indwelling ruminal probe methodology and effect of grain level on diurnal pH variation in dairy cattle

To evaluate a ruminal probe for recording diurnal variation in rumen pH, we fitted three ruminally fistulated cows with probes affixed to the inside of the cannula. Probes were connected to a data logger and readings were recorded hourly. The experiment was a Latin square design with cows fed three different diets: 50, 60, and 70% grain as a total mixed rations once daily. Feed offerings and refusals were recorded daily. The experimental period was 21 d. The first 6 d were for adaptation, followed by 5-d rotations through each of the diets. Daily probe readings were recorded at 4-, 6-, 8-, 12-, and 24-h intervals. At each interval, readings were recorded (precleaned), a sample of rumen fluid was taken, and pH was measured in the laboratory. As probes were removed from the rumen, probe ends were cleaned with 0.1 N HCl and reinserted into the rumen, and a reading was recorded (postcleaned). No protective pH probe shields were used in this experiment. There were no differences between pre- and postcleaned pH readings across cows for all diets. Mean time under pH 5.0, 5.5, and 6.0 were 0.2, 2, and 7.2 h, respectively. Diet affected length of time under a certain pH, only for readings under pH 6. Diurnal pH profiles were monophasic in nature. The degree of acidity increased after feeding and duration of nadir increased with increasing grain in the diet. Daily DMI increased but was highly variable within the first week after switching to the next higher grain level. These results indicate the use of an indwelling ruminal probe without a protective shield, cleaned, and calibrated daily can accurately measure diurnal variation of ruminal pH. In addition, transition to higher grain levels in the diet increases pH, duration of pH nadir, and daily DMI fluctuation.     

Effects of feeding a calf starter on molecular adaptations in the ruminal epithelium and liver of Holstein dairy calves

The objective of this study was to elucidate the effect of feeding a calf starter on the volatile fatty acid (VFA) profile in the rumen and on expression of genes involved in epithelial intracellular pH regulation, butyrate metabolism, and hepatic urea cycle during the weaning transition. Twenty Holstein bull calves were fed either milk replacer and hay (MR) or milk replacer, 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 the MR+S treatment were also fed starter ad libitum, and the energy intake of calves within blocks was maintained by supplementing the MR group with extra milk replacer that was equivalent to the energy intake from calf starter. Calves were killed 3 d after they consumed 680 g/d of calf starter for 3 consecutive days. Calves fed MR+S had higher VFA concentrations in the rumen (99.1±8.1 vs. 64.6±8.6 mM) and a higher molar proportion of butyrate (15.6±1.7 vs. 7.9±1.9%) than calves fed MR. Relative abundance of mRNA for monocarboxylate transporter isoform 1 was higher (1.45 vs. 0.53), and that of Na(+)/H(+) exchanger isoform 3 (0.37 vs. 0.82) and 3-hydroxy-3-methylglutaryl synthase isoform 1 (0.40 vs. 0.94) lower for the MR+S treatment compared with the MR treatment. In the liver, relative mRNA abundances of argininosuccinate synthetase isoform 1 (2.67 vs. 1.56), argininosuccinate lyase (1.44 vs. 0.99), and arginase isoform 1 (3.21 vs. 1.74) were greater for MR+S than for MR calves. Calf starter consumption appeared to increase fermentation in the rumen and affected expression of genes involved in cholesterol synthesis and intracellular pH regulation in ruminal epithelium, and those involved in urea cycle in the liver.     

Effect of milk allowance on concentrate intake, ruminal environment, and ruminal development in milk-fed Holstein calves

The aim of the present experiment was to test the hypothesis that a barley-based concentrate would induce an acidic ruminal environment in young calves and that increased milk allowance would alleviate this condition. Eight Holstein calves ruminally cannulated at d 7 ± 1 of age were used to study the effect of variation in barley-based starter concentrate intake induced by 4 different milk allowances (3.10, 4.84, 6.60, and 8.34 kg of milk replacer/d; 123 g of dry matter/kg of milk) on the ruminal environment, blood variables, and fore-stomach development from wk 2 to 5 of age. Twelve ruminal fluid samples were collected during a weekly 24-h sampling in 4 consecutive weeks. Blood samples were collected by venipuncture between 1200 and 1300 h on ruminal sampling days. Rumen papillae development and visceral organ mass were recorded at slaughter. A linear treatment × week effect was observed for concentrate intake, with the calves fed the lowest milk allowance having the fastest increase in concentrate intake whereby these calves reached the same ME intake in wk 5 compared with calves with the highest milk allowance. Effects on ruminal variables were dominated by week of sampling, with minor differences among treatments. Ruminal pH was below 5.5 for 5 to 13 h/d and all calves with concentrate intake above 20 g of dry matter/d were observed to have a daily ruminal pH minimum at pH 5.5 or lower. The ruminal concentration of total volatile fatty acids (VFA) increased from 71 to 133 ± 9 mmol/L in wk 2 to 5 and was characterized by a relatively high molar proportion of propionate, increasing from 34 to 40 mol/100 mol of VFA in wk 2 to 5. In addition, the presence of ethanol and propanol as well as numerous VFA esters points to a ruminal environment with a relatively high hydrogen pressure. Plasma glucose and insulin responded to the highest milk allowance in wk 2 to 4. Plasma VFA and ketone bodies increased with the lowest milk allowance in wk 4 to 5. At slaughter, empty wet weights of the rumen + reticulum and omasum as well as mass of digesta in these compartments were found to decrease linearly and perirenal fat was found to increase linearly with milk allowance, indicating that the milk allowance changed the body composition of the calves. Lengths of ruminal papillae in the atrium and ventral ruminal sac were not affected by treatment. We concluded that the ruminal environment of young calves fed a barley-based starter concentrate was characterized by a low ruminal pH and high VFA concentration regardless of the milk allowance.     

Technical note: effect of removal of microbial cells by centrifugation on peptide and alpha-amino nitrogen concentrations in ruminal fluid

We evaluated the effect of centrifuging rumen fluid prior to analysis on concentrations of alpha-amino N (AAN) and peptides. Rumen fluid was collected from steers fed grain-based diets at either various times after feeding or after dosing the rumens with solubilized casein. Fluid was either directly processed for peptide analysis by acidifying 10 ml of rumen fluid with 0.5 ml of 70% (wt/wt) perchloric acid, or first centrifuged at 500 x g for 20 min to remove protozoa and then at 30,000 x g for 15 min to remove bacterial cells prior to further processing. By removing microbial cells, intracellular AAN and peptides were not included in subsequent analyses. Concentrations of AAN were determinedusing an automated trinitrobenzene sulfonic acid assay, and peptides were determined as the increase in AAN following acid hydrolysis of the samples. When casein was not dosed, removal of microbial cells prior to analysis decreased concentrations of both AAN andpeptides, and the decrease was greater for AAN (2.2 mM)than for peptides (1.2 mM). Dosing with casein led to much higher concentrations of ruminal peptides and AAN. After casein dosing, decreases in AAN and peptidecon-centrations due to prior centrifugation (2.1 mM and 1.0 mM for AAN a nd pept ides, respectively) were similar to the decreases observed before the casein dosing. Results suggest that the contribution of intracellular AAN and peptides to the concentrations in ruminal fluid are relatively constant across broad ranges of dietary protein supply for cattle fed corn-based diets.     

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.     

Technical note: Ruminal cannulation technique in young Holstein calves: Effects of cannulation on feed intake, body weight gain, and ruminal development at six weeks of age

Ruminal cannulation techniques are frequently used to study fermentation in the ruminant forestomach. Unsatisfactory results with the traditionally applied procedure for cannulation of young calves stimulated the development of a simpler and more robust procedure; this procedure was tested for effects on performance traits and gross anatomy of the gastrointestinal tract compared with a control group not undergoing surgery. Five calves were ruminally cannulated at approximately 10 d of age and 5 matching calves were used as controls. All calves were fed milk replacer and a diet based on clover grass silage and sodium hydroxide-treated wheat. Ruminal fluid was collected from cannulated calves once weekly for 3 consecutive weeks. All calves were euthanized at 43 ± 3 d of age. No apparent adverse effects of cannulation were observed. Feed intake, BW gain, and gross anatomy of the gastrointestinal tract were not affected by cannulation. Minimum ruminal pH increased with sampling week, but average ruminal pH, total volatile fatty acids concentration, and volatile fatty acids proportions were not affected by sampling week. In conclusion, the implemented surgical technique was found to have no major effect on apparent animal health and performance traits, and the cannula proved useful for multiple samplings of ruminal contents in young calves.     

Changes in the rumen and colon microbiota and effects of live yeast dietary supplementation during the transition from the dry period to lactation of dairy cows

The first objective of this study was to evaluate the dynamics and their potential association with animal performance of the microbiota in both the rumen and colon of dairy cows as they move from a nonlactation to a lactation ration. The second objective was to assess the potential effects on the microbiota of live yeast supplementation. Twenty-one Holstein cows were split in 2 treatments consisting of 1 × 10¹⁰ cfu/d of live yeast (LY; n = 10) or no supplementation (control; n = 11) starting 21 d before until 21 d after calving. At 14 d before and 7 and 21 d after calving, samples of rumen and colon digesta were obtained from each cow using an endoscope. Total DNA was extracted and submitted to high-throughput sequencing. Shannon diversity index, in both the rumen and colon, was unaffected by LY; however, in the rumen it was lowest 7 d after calving and returned to precalving values at 21 d in milk, whereas in the colon it was greatest 14 d before calving but decreased after calving. In the rumen, LY supplementation increased the relative abundance (RA) of Bacteroidales (group UCG-001), Lachnospiracea (groups UCG-002 and UCG-006), and Flexilinea 14 d before calving, and increased RA of Streptococcus 21 d after calving compared with control cows. However, changes in the ruminal microbiota were more drastic across days relative to calving than as influenced by the dietary treatment, and the effect of LY in the colon was milder than in the rumen. The ruminal RA of several genera was associated with postcalving DMI, and that of Gastranaerophilales was the only order positively associated with milk yield. Several genera were positively correlated with feed efficiency, with Clostridiales (unclassified) being the only genus negatively associated with feed efficiency. In the colon, Prevotellaceae (group Ga6A1) was the only genus positively associated with feed efficiency. The ruminal RA of Prevotella 7 and Ruminobacter 14 d precalving was negatively correlated with dry matter intake and milk yield postcalving. The RA of Parabacteroides in the colon 14 d before calving was negatively correlated with milk yield, whereas the RA of Eggerthellaceae (unclassified) and Erysipelotrichaceae (groups c and unclassified) were positively correlated with feed efficiency. Interestingly, LY supplementation doubled the RA of Eggerthellaceae (unclassified) in the colon. It is concluded that microbial diversity in the rumen experiences a transient reduction after calving, whereas in the colon, the reduction is maintained at least until 21 d in milk. Most of the effects of LY on rumen microbiota were observed before calving, whereas in the colon, LY effects were more moderate but consistent and independent of the stage of production. The microbial community of the rumen after calving is more associated with feed intake, milk yield, and feed efficiency than that of the colon. However, the colon microbiota before calving is more associated with feed efficiency after calving than that of the rumen.     

Technical note: Evaluation of a wireless pulse oximeter for measuring arterial oxygen saturation and pulse rate in newborn Holstein Friesian calves

Pulse oximetry is a well-established technique in human and veterinary medicine. In farm animals, it could also be a useful tool for the detection of critical conditions relating to oxygen supply and the cardiovascular system. Among other uses, an innovative application could be the monitoring of fetuses during birth. This could help in the early identification of critical situations and support farmers and veterinarians in their decision to start obstetric or life-support interventions. Until now, however, its use in ruminant medicine was still limited to experimental applications. The objective of this study was to evaluate the accuracy of the Radius-7 Wearable Pulse CO-Oximeter (Masimo Corporation, Irvine, CA) for monitoring vital parameters in newborn calves. All measurements were conducted on animals in the lying down position. The sensor of the pulse oximeter was placed in the interdigital space of the calves' front legs and fixed with a homemade latex hoof cover. The pulsoximetric measurements of arterial oxygen saturation (SpO₂) in 40 newborn calves were compared with the corresponding results (SaO₂) from a portable blood gas analyzer (VetScan iStat1, Abaxis Inc., Union City, CA), which served as the reference. For this, an arterial blood sample was taken from the medial intermediate branch of the caudal auricular artery. In addition, the pulse rate was measured in 10 calves aged between 0 and 7 d with the pulse oximeter and simultaneously with a heart rate belt (Polar Equine Belt, Polar Electro Oy, Kempele, Finland) to determine their level of agreement. Spearman correlation coefficient for oxygen saturation was 93.8% for the pulse oximeter and the blood gas analyzer, and 97.7% for the pulse rate measured with the pulse oximeter and the heart rate belt. Bland-Altman plots revealed an overestimation of SaO₂ by 2.95 ± 6.39% and an underestimation of the pulse rate by ?0.41 ± 3.18 beats per minute compared with the corresponding reference methods. In summary, the pulse oximeter is suitable for continuous monitoring of arterial oxygen saturation and pulse in newborn Holstein Friesian calves. For practical use, purpose-built technical equipment is required to attach the sensor to the calves' legs.     

Repeated ruminal acidosis challenges in lactating dairy cows at high and low risk for developing acidosis: feed sorting

An experiment was conducted to determine whether the susceptibility of cows to ruminal acidosis influences feed sorting and whether feed sorting changes during a bout of ruminal acidosis. Eight ruminally cannulated cows were assigned to 1 of 2 acidosis risk levels: low risk (LR, mid-lactation cows fed a 60% forage diet) or high risk (HR, early lactation cows fed a 45% forage diet). As a result, diets were intentionally confounded with milk production to represent 2 different acidosis risk scenarios. Cows were exposed to an acidosis challenge in each of two 14-d periods. Each period consisted of 3 baseline days, a feed restriction day (restricting TMR to 50% of ad libitum intake), an acidosis challenge day (1-h meal of 4 kg of ground barley/wheat before allocating the TMR), and a recovery phase. Ruminal pH was measured continuously for the first 9 d of each period using an indwelling system. Feed and orts were sampled for 2 baseline days, on the challenge day, and 1 and 3 d after the challenge day for each cow and subjected to particle size analysis. The separator contained 3 screens (18, 9, and 1.18 mm) and a bottom pan to determine the proportion of long, medium, short, and fine particles, respectively. Sorting was calculated as the actual intake of each particle size fraction expressed as a percentage of the predicted intake of that fraction. All cows sorted against the longest and finest TMR particles and sorted for medium-length particles. Sorting was performed to a greater extent by the HR cows, and this sorting was related to low ruminal pH. Both HR and LR cows altered their sorting behavior in response to acidosis challenges. For the HR cows, severe acidosis was associated with increased sorting for the longer particles in the diet and against the shorter particles, likely to lessen the effects of the very low ruminal pH. These results suggest that feed sorting is affected by whether cows are at risk of acidosis. Furthermore, cows experiencing severe acidosis preferentially sort their feed to attenuate the effects of this condition.     

Structural growth, rumen development, and metabolic and immune responses of Holstein male calves fed milk through step-down and conventional methods

Structural growth, feed consumption, rumen development, metabolic response, and immune response were studied in Holstein calves fed milk through either a conventional method or a step-down (STEP) method. In the conventional method, calves (n = 20) were fed colostrum and then milk at a rate of 10% of their BW for the entire period of 44 d. In the STEP method, calves (n = 20) were given colostrum and then milk at a rate of 20% of their BW for 23 d, which was reduced (between d 24 to 28) to 10% of their BW for the remaining 16 d. The calves on both methods were weaned gradually by diluting milk with water between d 45 and 49. After weaning, feed consumption, structural growth, and body weight gain were monitored until calves were 63 d of age. At d 63, twelve calves (6/treatment) were euthanized and rumen papillae length, papillae width, rumen wall thickness, and emptied forestomach weight were recorded. At wk 4, 7, and 9, ruminal contents were collected to enumerate rumen metabolites. The STEP-fed calves consumed a greater amount of milk than conventionally fed calves during the pre-STEP (d 1 to 28), post-STEP (d 29 to 49), and preweaning (d 1 to 49) periods. Consumption of starter and hay was greater during the pre-STEP period and lesser during the post-STEP and postweaning (d 50 to 63) periods in calves on the conventional method than on the STEP method. Body weight gain and structural growth measurements of calves were greater on the STEP method than on the conventional method. A hypophagic condition caused by greater milk consumption depressed solid feed intake of STEP-fed calves during the pre-STEP period, and a hyperphagic response caused by a reduced nutrient supply from milk triggered their consumption of solid feed during the post-STEP and postweaning periods. Ruminal pH and concentrations of ammonia, total volatile fatty acids, acetate, propionate, butyrate, and plasma β-hydroxybutyrate were higher in calves on the STEP method and at weaning and postweaning (d 63) were lower in calves on the conventional method. Emptied weight of the forestomach, rumen wall thickness, papillae length, papillae width, and papillae concentration were higher in calves on the STEP method than in those on the conventional method. Blood glucose was lower, and blood urea nitrogen and β-hydroxybutyrate at weaning and postweaning were higher in STEP-fed calves. Serum IgG, IgA, and triglycerides for 1, 2, and 3 wk of age were higher in calves on the STEP method than in those on the conventional method. In conclusion, greater feed consumption, BW gain, and structural growth, and a more metabolically and physically developed rumen were observed in calves on the STEP method than in those on the conventional method.