Investigating nutritional strategies during a rest period to improve health, growth, and behavioral outcomes of transported surplus dairy calves.

The objective of this study was to investigate the effects of feeding surplus dairy calves a milk replacer (MR) or one of 2 different oral rehydration solutions (ORS) during a mid-transportation rest period on metabolic and clinical health indicators, growth, and behavioral outcomes after arrival to a calf-raising facility. Surplus dairy calves (n = 128) were transported in 4 cohorts from February to July 2022 for 12 h to a holding facility, rested for 8 h, then transported for an additional 6 h to a calf-raising facility. Upon arrival to the holding facility, calves were randomly assigned to 1 of 3 treatments: MR (n = 43), a high sodium ORS developed for diarrhea (ORS-D; n = 43), or a high potassium ORS developed for transportation (ORS-T; n = 42). The exact age of calves at transportation was unknown, however all calves were under 14 d of age. Calf body weight at enrollment was 43.9 ± 5.9 kg, 43.7 ± 6.5 kg, and 45.0 ± 4.5 kg for calves fed MR, ORS-D, and ORS-T, respectively. Calves were fed 2.0 L of their treatment twice, once upon arrival and once before leaving the holding facility. At unloading and reloading at the holding facility, calves were weighed and blood sampled. Calves were also health scored at unloading at the holding facility. After arrival at the calf-raising facility, calves were weighed, health scored, and blood sampled. Blood samples were collected at 24 and 48 h and body weight (BW) was recorded at 24 h, 48 h, 72 h, 5 d, 7 d, 14 d, and at 8 wks after arrival to the calf-raising facility. Calves were also health scored daily for 14 d, which included fecal consistency scoring and evaluating the presence or absence of respiratory disease. Lying time, lying bouts, and activity index were measured during transportation and from 3 d relative to transportation using accelerometers. At arrival to the calf-raiser, calves fed ORS-D had higher concentrations of NEFA and BHB than calves fed MR. Furthermore, calves fed ORS-T had higher concentrations of BHB at arrival to the calf raiser compared with calves fed MR. In the 14 d after arrival to the calf-raiser, there was evidence that calves fed ORS-T had a higher proportion of days with diarrhea and respiratory disease compared with those fed MR. During transportation, calves fed ORS-T had a lower activity index than calves fed MR, suggesting that ORS-T calves had lower overall activity. Additionally, on the day of transportation (d 0), ORS-T and ORS-D calves had a lower activity index than calves fed MR. There were no treatment effects on growth outcomes. The results of this study suggest that feeding MR rather than an ORS during a mid-transportation rest period could minimize fat mobilization and can potentially improve diarrhea and respiratory disease but does not affect growth outcomes after arrival to calf-raisers.

Effects of supplementing colostrum beyond the first day of life on growth and health parameters of preweaning Holstein heifers.

The preweaning period for a dairy calf is characterized by high morbidity and mortality rates, leading to financial losses for producers. Identifying strategies to improve the health and welfare of calves while reducing antimicrobial use continues to be crucial to the success of the dairy industry. The objective of this study was to determine the effects of feeding colostrum replacer (CR) to dairy heifer calves beyond d 1 of life on growth, serum IgG, the incidence of diarrhea and bovine respiratory disease (BRD), and the risk of mortality in the preweaning period. At birth, Holstein heifer calves (n = 200; 50/treatment) weighing 40.7 ± 0.35 kg (mean ± SE) were fed 3.2 L of CR (205 g IgG/feeding) at 0 h and 12 h of life. Calves were then randomly assigned to 1 of 4 treatments: 450 g of milk replacer (MR) from d 2 to 14 (control, CON), 380 g of CR + 225 g of MR from d 2 to 3, then 450 g of MR from d 4 to 14 (transition, TRAN), 45 g of CR + 450 g of MR from d 2 to 14 (extended, EXT); or 380 g of CR + 225 g of MR from d 2 to 3, then 45 g of CR + 450 g of MR from d 4 to 14 (transition + extended, TRAN+EXT). Each treatment was reconstituted to 3 L and fed twice daily. All CR treatments were fed using bovine-derived CR containing 27% IgG. From d 15 to 41, all calves were fed 600 g of MR reconstituted to 4 L twice daily. Body weight was recorded at birth and every 7 d until study completion on d 49. Blood samples were taken daily until d 7 to evaluate serum IgG and then every 7 d until d 49. A health assessment was performed daily to evaluate calves for BRD and diarrhea. Data were analyzed using mixed linear regression, mixed logistic regression, and survival analysis models in SAS 9.4. Serum IgG concentrations were not affected by treatment for the study period. The EXT and TRAN+EXT groups had greater average daily gain (ADG) from d 7 to 14 (0.14 kg/d) and the TRAN group had greater ADG from d 14 to 21 (0.11 kg/d), compared with CON. There was no association of treatment with the odds or the duration of a diarrhea bout. However, provision of CR to the TRAN and EXT calves was associated with a reduced hazard of diarrhea compared with CON calves. Furthermore, TRAN and EXT calves have a lower hazard of mortality compared with CON calves, with TRAN and EXT calves had a 2.8- and 3.8-times lower hazard of mortality, respectively. Our findings suggest that the supplementation of CR to dairy calves positively affects ADG, and reduces the hazard of diarrhea and mortality during the preweaning period. Future research should look to further refine the supplementation strategy of CR to calves and explore the mechanism of action.

The development and validation of a milk feeding behavior alert from automated feeder data to classify calves at risk for a diarrhea bout: A diagnostic accuracy study.

The objective of this diagnostic accuracy study was to develop and validate an alert to identify calves at risk for a diarrhea bout using milk feeding behavior data (behavior) from automated milk feeders (AMF). We enrolled Holstein calves (n = 259) as a convenience sample size from 2 facilities that were health scored daily preweaning and offered either 10 or 15 L/d of milk replacer. For alert development, 132 calves were enrolled and the ability of milk intake, drinking speed, and rewarded visits collected from AMF to identify calves at risk for diarrhea was tested. Alerts that had high diagnostic accuracy in the alert development phase were validated using a holdout validation strategy of 127 different calves from the same facilities (all offered 15 L/d) for -3 to 1 d relative to diarrhea diagnosis. We enrolled calves that were either healthy or had a first diarrheal bout (loose feces ≥2 d or watery feces ≥1 d). Relative change and rolling dividends for each milk feeding behavior were calculated for each calf from the previous 2 d. Logistic regression models and receiver operator curves (ROC) were used to assess the diagnostic ability for relative change and rolling dividends behavior relative to alert d) to classify calves at risk for a diarrhea bout from -2 to 0 d relative to diagnosis. To maximize sensitivity (Se), alert thresholds were based on ROC optimal classification cutoff. Diagnostic accuracy was met when the alert had a moderate area under the ROC curve (≥0.70), high accuracy (Acc; ≥0.80), high Se (≥0.80), and very high precision (Pre; ≥0.85). For alert development, deviations in rolling dividend milk intake with drinking speed had the best performance (10 L/d: ROC area under the curve [AUC] = 0.79, threshold ≤0.70; 15 L/d: ROC AUC = 0.82, threshold ≤0.60). Our diagnostic criteria were only met in calves offered 15 L/d (10 L/d: Se 75%, Acc 72%, Pre 92%, specificity [Sp] 55% vs. 15 L/d: Se 91%, Acc 91%, Pre 89%, Sp 73%). For holdout validation, rolling dividend milk intake with drinking speed met diagnostic criteria for one facility (threshold ≤0.60, Se 86%, Acc 82%, Pre 94%, Sp 50%). However, no milk feeding behavior alerts met diagnostic criteria for the second facility due to poor Se (relative change milk intake -0.36 threshold, Se 71%, Acc 70%, and Pre 97%). We suggest that changes in milk feeding behavior may indicate diarrhea bouts in dairy calves. Future research should validate this alert in commercial settings; furthermore, software updates, support, and new analytics might be required for on-farm application to implement these types of alerts.

Activity behaviors and relative changes in activity patterns recorded by precision technology were associated with diarrhea status in individually housed calves.

The objective of this case-control study was to quantify any association of daily activity behaviors and relative changes in activity patterns (lying time, lying bouts, step count, activity index) with diarrhea status in preweaning dairy calves. Individually housed calves sourced from auction were health-scored daily for signs of diarrhea (fecal consistency loose or watery for 2 consecutive days) for the 28 d after arrival. Calves with diarrhea were pair-matched with healthy controls (n = 13, matched by arrival date, arrival weight, and diagnosis days to diarrheic calves). Mixed linear regression models were used to evaluate the association of diarrhea status, and the diarrhea status by day interaction with activity behaviors (d -3 to d 4) and relative changes in activity patterns (d -3 to d 4) relative to diagnosis of a diarrhea bout. The serum Brix percentage at arrival and daily temperature-humidity index from the calf barn were explored as quantitative covariates, with day as a repeated measure. The baseline for relative changes in activity patterns was set at 100% on d 0. Diarrheic calves were less active; they averaged fewer steps (119.1 ± 18.81 steps/d) than healthy calves (227.4 ± 18.81 steps/d, LSM ± SEM). Diarrheic calves also averaged lower activity indices (827.34 ± 93.092 daily index) than healthy calves (1,396.32 ± 93.092 daily index). We also found also a diarrhea status by day interaction for lying time on d -3, with diarrheic calves spending more time lying (20.80 ± 0.300 h/d) than healthy calves (19.25 ± 0.300 h/d). For relative changes in activity patterns, a diarrhea status by day interaction was detectable on d -2, where diarrheic calves had greater relative changes in step counts (diarrhea 634.85 ± 87.581% vs. healthy 216.51 ± 87.581%) and activity index (diarrhea 316.83 ± 35.692% vs. healthy 150.68 ± 35.692%). Lying bouts were not associated with diarrhea status. These results show that diarrheic calves were more lethargic, and they had relative changes in activity patterns 2 d before clinical signs of diarrhea. Future research should explore the potential of an activity alert that positively indicates an individually housed calf at risk for a diarrhea bout using deviations from relative changes in individual calf activity patterns.

Evaluation of a point-of-care meter for measuring glucose concentrations in dairy calves: A diagnostic accuracy study.

The objective of this cross-sectional, diagnostic accuracy study was to validate a human blood glucose meter (Contour Next One Meter, Ascensia Diabetes Care) for accuracy and precision when measuring blood glucose, and for diagnostic accuracy for hypoglycemic status in dairy calves using whole blood and blood plasma. A total of 49 male dairy calves [body weight (BW): 46.3 ± 0.8 kg] had jugular catheters placed within 75 min after birth. Thereafter, blood was withdrawn from the catheter at specific time points (-10, 10, 20, 30, 45, 60, 90, 120, 180, 240, 360, 480, and 600 min) relative to the first and second colostrum feedings (2 h 15 min and 14 h 5 min postnatal; feeding rate: 7% of BW wt/wt). The reference standard method for plasma glucose concentration was determined colorimetrically and in duplicate using the glucose oxidase-peroxidase reaction. Data were assessed for agreement between the glucose meter and the reference standard using Lin's concordance correlation coefficients (CCC), coefficients of determination (precision), and Bland-Altman plots. In addition, a mixed linear regression model was built using the reference method as the outcome, with the glucose meter and repeated measures of time as the explanatory variables and calf as a random effect. The sensitivity (Se), specificity (Sp), and area under the curve (AUC) for the glucose meter using calf whole-blood and plasma were calculated at a threshold of <4.44 mmol/L to determine hypoglycemia. The precision (CCC = 0.95, R2 = 0.93) and accuracy (AUC = 0.98) of the glucose meter were very high when used on 1,303 blood plasma samples. Youden's index revealed a threshold of <4.45 mmol/L for the glucose meter when used with plasma, leading to Se of 94.2% and Sp of 91.9%, with 92.5% of samples being correctly classified, suggesting high diagnostic accuracy. When using whole blood, precision (CCC = 0.85 and R2 = 0.73) and accuracy (AUC = 0.92) were high when used on 476 samples. Youden's index revealed a threshold of <4.95 mmol/L for the glucose meter when used with whole calf blood, leading to Se of 95.6% and Sp of 80.3%, with 84.7% of samples being correctly classified, suggesting high diagnostic accuracy for use on farm. In summary, this glucose meter was validated for measuring calf blood glucose using both plasma and whole blood. This meter can measure glycemic status in calves and may be useful for clinical and on-farm use to make intervention decisions.

Evaluation of an infrared thermography camera for measuring body temperature in dairy calves.

The objective of this diagnostic accuracy study was to validate an infrared thermography (IRT) camera and its software (FLIR One, FLIR, Global) for accuracy and precision for ocular temperature readings to serve as a proxy for rectal temperature in commercially housed calves. A total of 318 male Holstein calves were enrolled into this study from the day of arrival to a calf rearing facility until 14 d later. Researchers took an ocular temperature reading using an IRT camera, and a rectal temperature on each calf each day in the morning. The reference standard method for body temperature in the calves was rectal temperature. We assessed the data for agreement between the IRT and the reference standard using Pearson correlations by calf (accuracy), coefficients of determination (precision), and Bland-Altman plots for bias. In addition, a logistic regression model was built using the reference method as the outcome, with IRT as an explanatory variable to assess the diagnostic accuracy of IRT as an indicator of fever (rectal temperature ≥39.5°C). There was a negligible correlation between the IRT readings and rectal temperature (r = 0.22) and the coefficient of determination for IRT to predict rectal temperature was negligible (R2 = 0.05), suggesting poor precision. The average mean difference between the IRT data and rectal temperature was 0.55°C, and the differences between IRT and rectal formed a linear line around the mean difference, suggesting the Bland-Altman analyses showed proportional error and bias. The optimal probability cut-off for IRT readings for fever was at 39.5°C, and had a receiver operating characteristic area under the curve of 0.67, a sensitivity of 61%, a specificity of 71%, and 78% (3,134/4,427) of the samples were correctly labeled as either having a fever or not using IRT readings. In summary, the IRT camera and software were not validated for serving as a proxy for rectal temperature in commercially housed calves due to poor precision, and proportional error partially explained by ambient environmental conditions. We suggest that this infrared thermography system should not replace rectal temperature readings for use in commercially housed calves.

Association of a green tea extract with serum immunoglobulin G status and neonatal vitality in newborn dairy calves.

Poor vigor at birth has been associated with reduced IgG absorption from colostrum and a reduced vitality in neonatal dairy calves. Some natural compounds, such as green tea extract, may improve vitality in compromised calves. The objective of this randomized controlled trial was to evaluate the potential of supplementing a green tea extract (15 mL) to calves to improve vigor and activity behavior for the first 72 h postnatal. Also, this study aimed to investigate the influence of green tea extract supplementation on calf serum IgG concentration and the apparent efficiency of absorption (AEA) of colostral IgG. Holstein calves (n = 24) weighing 42.49 ± 1.07 kg postnatal received a complete random assignment at 3 h of one 15-mL dose of green tea extract (Calf Perk, TechMix) or distilled water orally before tube feeding colostrum replacer (Premolac Plus IgG, Zinpro) at 4 h postnatal. Two observers assessed for calving time and dystocia by live video stream to retrieve all calves within 2 h postnatal. One veterinarian performed a baseline vigor assessment based on heart rate and response to stimuli on all calves at 2.5 h, before colostrum feeding at 3.75 h, as well as at 24, 48, and 72 h postnatal. Calf blood samples were taken to assess total IgG by radial immunodiffusion assay at 2.5, 6, 12, 24, 48, and 72 h. Calf vitality was also observed continuously by video for all calves to determine whether treatment was associated with attempts to stand, lying time, and exploration of their pen environment for the first 24 h. We used an ordinal logistic model to evaluate the odds of green tea extract improving a calf's vigor category from 2.5 h postnatal to 72 h of age. Vigor score was categorized as abnormal (≤4), average (5), or alert (≥6), with hour as a fixed effect. We also ran mixed linear models to evaluate the effect of extract on total IgG and AEA, with time and dystocia as fixed effects. Five dystocia calves were enrolled (2 control, 3 extract), but assistance was minor (e.g., manual assistance and all were assisted within 1 h). Baseline vigor scores and baseline total IgG were not different between groups. Vigor score category was not associated with green tea extract supplementation (odds ratio 1.17; 95% CI: 0.43-3.15) but increased with time compared with controls. We observed no association of treatment with total IgG or AEA in the calves, suggesting green tea extract does not compromise IgG absorption. Calf vitality, lying behavior, and exploratory behavior were not associated with green tea extract treatment. Our findings suggest that green tea extract supplementation does not affect AEA and serum IgG concentration in calves. Future research should evaluate whether green tea extract improves vitality in calves experiencing severe dystocia.

Effect of supplementation with algae ß-glucans on performance, health, and blood metabolites of Holstein dairy calves.

Studies have shown that ß-glucans extracted from the cell wall of cereals, algae, and yeasts have been associated with improved immune function. However, it is unknown whether algae ß-glucan supplementation affects the performance, blood metabolites, or cell counts of immune cells in dairy calves. The objective of this randomized clinical trial was to evaluate whether supplementation of ß-glucans to milk replacer in dairy calves fed 6 L/d improved growth performance and fecal status and altered the blood metabolite profile. In this trial, we enrolled Holstein calves (n = 34) at birth (body weight 36.38 ± 1.33 kg; mean ± standard deviation) to receive, from 1 d of age, either 2 g/d algae ß-glucans mixed into 6 L/d of milk replacer (22.4% crude protein and 16.2% fat) or an unsupplemented milk replacer (control). The calves were blocked in pairs according to birth weight, sex, and date of birth (up to 5 d difference). Calves were housed individually, and calf starter (24.7% crude protein and 13.9% neutral detergent fiber) was offered ad libitum based on orts of the previous day until 56 d of age (end of the trial). Body weight was measured weekly, and health checks and daily fecal consistency were evaluated daily in every calf by the same observer. Calves with 2 consecutive days of loose feces that sifted through bedding were considered diarrhea positive. We used a linear mixed effects model to evaluate the effects of ß-glucan supplementation fed during the preweaning period on performance (average daily gain), final weight, feed efficiency (FE), white blood cell count, and selected blood metabolites, repeated by time. A generalized linear mixed effects model was also run to evaluate the likelihood of a diarrhea bout in the first 28 d of life, controlling for the calf as the subject with a logistic distribution. We included age, serum total protein at 48 h, and birth weight as covariates. At 56 d, ß-glucan-supplemented calves weighed more than control calves (56.3 vs. 51.5 kg). Treatment had no effect on total starter intake, but there was a treatment by age interaction for FE, with greater FE for ß-glucan-supplemented calves in wk 3 and 5 of age. There was only a tendency for average daily gain to be greater in supplemented calves than in control calves for the duration of the study. Furthermore, control calves had 14.66 [95% confidence interval (95% CI): 9.87-21.77] times greater odds of having a diarrheal bout than ß-glucan-supplemented calves. Control calves had 12.70 (95% CI: 8.82-18.28) times greater odds of having an additional day with an abnormal fecal score compared with ß-glucan-supplemented calves, suggesting that supplementation ameliorated diarrhea severity. We found no association of treatment with concentrations of serum total protein, albumin, creatinine, or glucose during the preweaning period. Our findings suggest that dietary supplementation of 2 g/d of algae ß-glucans to milk replacer improved fecal status and may affect growth, as evidenced by a higher weaning weight, compared with control calves. Future studies should explore the effect of algae ß-glucans on lower-gut physiology and digestibility in dairy calves.

Feeding behavior and activity levels are associated with recovery status in dairy calves treated with antimicrobials for Bovine Respiratory Disease.

Calves with Bovine Respiratory Disease (BRD) have different feeding behavior and activity levels prior to BRD diagnosis when compared to healthy calves, but it is unknown if calves who relapse from their initial BRD diagnosis are behaviorally different from calves who recover. Using precision technologies, we aimed to identify associations of feeding behavior and activity with recovery status in dairy calves (recovered or relapsed) over the 10 days after first antimicrobial treatment for BRD. Dairy calves were health scored daily for a BRD bout (using a standard respiratory scoring system and lung ultrasonography) and received antimicrobial therapy (enrofloxacin) on day 0 of initial BRD diagnosis; 10-14 days later, recovery status was scored as either recovered or relapsed (n = 19 each). Feeding behaviors and activity were monitored using automated feeders and pedometers. Over the 10 days post-treatment, recovered calves showed improvements in starter intake and were generally more active, while relapsed calves showed sickness behaviors, including depressed feed intake, and longer lying times. These results suggest there is a new potential for precision technology devices on farms in evaluating recovery status of dairy calves that are recently treated for BRD; there is opportunity to automatically identify relapsing calves before re-emergence of clinical disease.

Daily behavioral measures recorded by precision technology devices may indicate bovine respiratory disease status in preweaned dairy calves.

Precision technology devices can measure and detect relative changes in an animal's behavior to possibly create alerts to intervene and to administer treatments. However, the association of relative changes in daily feeding and activity behaviors in calves with bovine respiratory disease (BRD) status is still largely unexplored. The objective of this case-control study was to determine if daily behavioral patterns of preweaning dairy calves (measured by precision technologies) change before BRD diagnosis. This case-control study enrolled 33 pairs of calves (33 BRD calves matched by age, sex, and birthdate to 33 controls) health scored daily for BRD for the preweaning period (until 50 d on the automated feeder). A pedometer (IceQube, Ice Robotics) was attached to the left rear leg to track activity (lying time, lying bouts, total steps, and acceleration activity index). At 3.0 ± 2.0 d of age, calves were trained to use the automated feeder, which recorded milk and calf starter intake, drinking speed, and feeder visits. Calves were allotted a maximum 10 L/d of milk replacer (Cow's Match, Land O'Lakes Animal Milk Products Co.; 140 g/L) and starter (Special Calf Starter and Grower, Baghdad Feeds), ad libitum. Calves were scored daily for signs of BRD using the Wisconsin scoring system, and their lungs were scored with ultrasonography twice weekly. Outward signs of BRD as defined by the Wisconsin scoring system and an area of consolidated lung ≥3.0 cm2 was identified as BRD (d 0). Relative changes in daily behaviors were calculated using d -5 before BRD diagnosis as the baseline for each calf for each behavior. Linear mixed models were used to investigate the association of BRD status with feeding behaviors, activity, and relative changes in calf behavior over a 5-d period before diagnosis, as well as the BRD status × day interaction. Calves with a BRD bout were diagnosed at an average age of 33.0 ± 9.0 d (mean ± standard deviation) and weighed 56.1 ± 9.7 kg. Over the period, BRD calves had reduced milk and starter intake, greater lying times, and fewer lying bouts, step counts, and activity indices when compared with healthy calves. Furthermore, there was a BRD status × day interaction for relative changes in unrewarded visits, and relative changes in calf starter intake. Specifically, BRD calves had a decline in relative changes in their unrewarded visits on d -4, -2, -1, and 0 compared with healthy calves. This study suggests that there is the potential to use feeding and activity behaviors to identify BRD development in preweaning calves. However, utilization of an animals' daily behavioral patterns in real time is fundamental for developing disease detection algorithms, thus we suggest relative changes in unrewarded visits may be useful for algorithm development when d -5 is used as a baseline. Future research should investigate the potential of feeding behavior and activity levels collectively to indicate BRD status in calves using machine learning techniques.

Nutraceutical intervention with colostrum replacer: Can we reduce disease hazard, ameliorate disease severity, and improve performance in preweaned dairy calves?

The objective of this randomized clinical trial was to determine if an intervention with colostrum replacer (CR; Premolac Plus, Zinpro; 125 g/d fed for 3 d) or a placebo milk replacer (Cows Match, Land O'Lakes Inc.; 125 g/d for 3 d) following a triggered alarm could ameliorate disease bouts in dairy calves. The alarm was set to detect negative deviations of milk intake (20% reduction) or drinking speed (30% reduction) in relation to a calf's 12-d rolling average feeding behavior. Calves were enrolled on this study (n = 42 CR, n = 42 placebo) when they triggered an alarm from d 14 to 50 on the feeder before weaning. Once calves were enrolled, calves received a bottle of either 125 g of CR or milk replacer mixed with 1 L of water for 3 d. Calves were enrolled on the automated milk feeder at age 4.0 ± 2.0 d (mean ± SD), were offered 10 L/d of the same milk replacer fed as a placebo, and had ad libitum access to calf starter measured by automated feeders. Weaning began at d 50, with 50% reduction of milk replacer across 14 d, and an additional 20% reduction for another 7 d before weaning at d 70. Calves were health scored from birth to 2 wk postweaning (88 ± 2.0 d of age) daily for bovine respiratory disease (BRD) and diarrhea, and were weighed and scored using lung ultrasonography twice weekly. A BRD score ≥5 and an area of consolidated lung ≥3.0 cm2 was considered BRD positive; diarrhea was defined as a watery fecal consistency that sifted through the bedding. The effect of CR on BRD and diarrhea likelihood were calculated with logistic models; the 7 d before and after an alarm were fixed effects, with milk intake as a covariate. A Cox proportional hazards model evaluated the effect of CR on the hazard of being positive for BRD and diarrhea following enrollment, with sex as a fixed effect in the diarrhea model. The effect of CR on average daily gain the week following intervention was evaluated with linear modeling with repeated measures, and birth date was a random effect. After intervention, placebo calves had 1.64 (95% CI: 1.11-2.43) times greater odds of having a BRD bout compared with CR calves for the 7 d following intervention. Moreover, placebo calves had 1.50 (95% CI: 1.11-2.08) times greater odds of having lobar lung consolidation in the 7 d after intervention than CR calves. No difference was found in the likelihood of diarrhea in the 7 d after intervention (odds ratio: 0.91; 95% CI: 0.71-1.16). However, placebo calves had a 2.38 (95% CI: 1.30-4.33) times greater hazard of incurring BRD in the 14 d after intervention. Average daily gain was not associated with an intervention with CR for the 7 d after intervention (placebo: 0.73 ± 0.07 kg/d; CR: 0.70 ± 0.08 kg/d; LSM ± SEM). These results suggest CR may reduce BRD likelihood but did not influence growth in calves offered high allowances of milk. Future research should examine which properties of CR ameliorate BRD in calves.

Supplementation of lysolecithin in milk replacer for Holstein dairy calves: Effects on growth performance, health, and metabolites.

Lysolecithin is an antiinflammatory emulsifier associated with improved apparent digestibility of total dietary fat and improved feed efficiency in dairy cattle. However, it is unknown if lysolecithin (LYSO) improves performance in calves. Moreover, since many conventional milk replacers use vegetable-sourced fat (e.g., palm oil), nutrient absorption and fecal score may be affected in neonatal calves. Thus, the objective of this study was to evaluate the effects of LYSO supplemented in milk replacer on performance, metabolites, and gut health of preweaned dairy calves. Holstein calves (n = 32) with adequate passive transfer were assigned in pairs (16 blocks) balanced by birth weight, date of birth, and sex at 1 d of age to randomly receive either LYSO (mixed in 2 milk replacer feedings at a rate of 4 g/d Lysoforte, Kemin Industries Inc., Des Moines, IA) or a milk replacer control (nothing added). Both treatments were fed 6 L/d milk replacer [22.5% crude protein, 16.2% crude fat (vegetable oil fat source) on a dry matter basis with 14% solids] by bucket in 2 daily feedings for 56 d. Calves were individually housed in wooden hutches and offered a commercial calf starter (24.6% crude protein and 13.9% neutral detergent fiber) and water by bucket ad libitum. Feed refusals and calf health was assessed daily. Weights and blood metabolites (glucose, total serum protein, albumin, creatinine, triglycerides, and cholesterol) were sampled weekly, and calves completed the study before weaning at 56 d of age. The effects of LYSO on calf average daily gain, feed efficiency, and blood metabolites were evaluated using a linear mixed model with time as a repeated measure, calf as the subject, and block as a random effect in SAS (SAS Institute Inc., Cary, NC). The effect of LYSO to improve the odds of abnormal fecal score was evaluated using a logistic model. Supplementation of LYSO increased average daily gain (control 0.28 ± 0.03 kg; LYSO 0.37 ± 0.03 kg; least squares means ± standard error of the mean) and increased feed efficiency (gain-to-feed; control 0.25 ± 0.03; LYSO 0.32 ± 0.03). Similarly, LYSO calves had a higher final body weight at d 56 (control 52.11 ± 2.33 kg; LYSO 56.73 ± 2.33 kg). Interestingly, total dry matter intake was not associated with LYSO despite improved average daily gain (total dry matter intake control 1,088.7 ± 27.62 g; total dry matter intake LYSO 1,124.8 ± 27.62 g). Blood glucose, albumin, creatinine, triglycerides, and cholesterol were not associated with LYSO. Indeed, only total serum protein had a significant interaction with LYSO and age at wk 5 and 6. Moreover, control calves had a 13.57 (95% confidence interval: 9.25-19.90) times greater odds of having an abnormal fecal score on any given day during the diarrhea risk period from d 1 to 28. The inclusion of LYSO as an additive in milk replacer in a dose of 4 g/d may improve performance, and calf fecal score, preweaning. Further research should investigate the mechanisms behind the effects of LYSO on fat digestibility in calves fed 6 L/d of milk replacer with vegetable-sourced fat.

A comprehensive evaluation of microchips to measure temperature in dairy calves.

Elevated temperature is often an indicator of an immune response and used in the diagnosis of illness in dairy calves; however, measuring rectal temperature is labor intensive and often not measured daily on the farm. The objective of this study was to measure body temperature using a microchip and determine an appropriate implant site that would passively read body temperature in dairy calves. First, the precision of the temperature microchips and the rectal thermometer were tested ex vivo. Then, Holstein bull calves (n = 12) at 14 ± 12 d (mean ± SD) of age were implanted with microchips subcutaneously by the scutulum of the ear, subcutaneously in the upper scapula (SCAP), and intramuscularly in the trapezius muscle of the neck. One week after implantation, a temperature reading was taken for every microchip implant site using a radiofrequency ID (RFID) reader, as well as rectally and in the tympanic membrane using a digital thermometer every 60 min for 24 h in each calf (hereafter, the hourly study). Additionally, microchip readings and rectal temperatures were taken daily at 0800 h from 8 wk of age (n = 9; 57 ± 12 d of age) until 2 wk postweaning for a subset of the bull calves used in the hourly study (hereafter, the daily study). In the ex vivo trial, the microchip readings were very highly correlated with the rectal thermometer (r = 0.96), and the average coefficient of variation between microchip readings was very low (0.12 ± 0.03%). The relationships between the microchip readings within ear, SCAP, and neck and rectal and tympanic temperatures were analyzed using Pearson correlations and Bland-Altman plots. The ear and neck readings were strongly correlated for the hourly study [individual animal correlation; median (Q1, Q3), r = 0.78 (0.73, 0.84)] and for the daily study [r = 0.79 (0.73, 0.89)] across calves. However, rectal temperature was not significantly correlated with ear, SCAP, neck, or tympanic temperature for the daily and hourly studies. Results suggest that temperature microchips measure temperature appropriately, but temperature is dependent upon the implant site in calves, and temperature measured at ear, SCAP, and neck implant sites cannot be used to estimate rectal temperature. Future research should determine thresholds for fever that are specific to implant sites in calves.

Homophily around specialized foraging underlies dolphin social preferences.

Individuals often associate socially with those who behave the same way. This principle, homophily, could structure populations into distinct social groups. We tested this hypothesis in a bottlenose dolphin population that appeared to be clustered around a specialized foraging tactic involving cooperation with net-casting fishermen, but in which other potential drivers of such social structure have never been assessed. We measured and controlled for the contribution of sex, age, genetic relatedness, home range and foraging tactics on social associations to test for homophily effects. Dolphins tended to group with others having similar home ranges and frequency of using the specialized foraging tactic, but not other traits. Such social preferences were particularly clear when dolphins were not foraging, showing that homophily extends beyond simply participating in a specific tactic. Combined, these findings highlight the need to account for multiple drivers of group formation across behavioural contexts to determine true social affiliations. We suggest that homophily around behavioural specialization can be a major driver of social patterns, with implications for other social processes. If homophily based on specialized tactics underlies animal social structures more widely, then it may be important in modulating opportunities for social learning, and therefore influence patterns of cultural transmission.

Transport of valine across the small intestinal epithelium in pigs fed different valine levels and Bacillus subtilis.

Mutants of Bacillus subtilis overproducing valine (B. subtilis VAL) could be an approach to supply pigs dietary valine (Val). In the study, 18 gilts were fed: (i) negative diet with a standardized ileal digestible (SID) Val:Lys of 0.63:1 (Neg); (ii) Neg added B. subtilis VAL (1.28 × 1011  cfu/kg as-fed) or; (iii) Neg added L-Val to a Val:Lys of 0.69:1. Using the Ussing chamber method, the study aimed to investigate whether (i) the diets affect intestinal transport of additions of 0, 5, 10 or 20 mmol Val/L from the mucosal to the serosal side and (ii) the B. subtilis VAL contributes to a net transport of Val produced in situ. The results showed that the Isc (ΔIscVal ) and release of Val to the serosal side solution (Srel ; µmol cm-2  min-1 ) increased with Val addition (linear and quadratic, p < .0001) but was similar for 5, 10 or 20 mmol Val/L and not affected by diet. No net transport of in situ produced Val by B. subtilis VAL was detected. In conclusion, feeding a Val-deficient diet with or without B. subtilis VAL or a Val sufficient diet did not affect the Val transport across intestinal epithelia. No in situ Val production by B. subtilis VAL was observed in the Ussing chambers.

Evaluation of in situ valine production by Bacillus subtilis in young pigs.

Mutants of Bacillus subtilis can be developed to overproduce Val in vitro. It was hypothesized that addition of Bacillus subtilis mutants to pig diets can be a strategy to supply the animal with Val. The objective was to investigate the effect of Bacillus subtilis mutants on growth performance and blood amino acid (AA) concentrations when fed to piglets. Experiment 1 included 18 pigs (15.0±1.1 kg) fed one of three diets containing either 0.63 or 0.69 standardized ileal digestible (SID) Val : Lys, or 0.63 SID Val : Lys supplemented with a Bacillus subtilis mutant (mutant 1). Blood samples were obtained 0.5 h before feeding and at 1, 2, 3, 4, 5 and 6 h after feeding and analyzed for AAs. In Experiment 2, 80 piglets (9.1±1.1 kg) were fed one of four diets containing 0.63 or 0.67 SID Val : Lys, or 0.63 SID Val : Lys supplemented with another Bacillus subtilis mutant (mutant 2) or its parent wild type. Average daily feed intake, daily weight gain and feed conversion ratio were measured on days 7, 14 and 21. On day 17, blood samples were taken and analyzed for AAs. On days 24 to 26, six pigs from each dietary treatment were fitted with a permanent jugular vein catheter, and blood samples were taken for AA analysis 0.5 h before feeding and at 1, 2, 3, 4, 5 and 6 h after feeding. In experiment 1, Bacillus subtilis mutant 1 tended (P<0.10) to increase the plasma levels of Val at 2 and 3 h post-feeding, but this was not confirmed in Experiment 2. In Experiment 2, Bacillus subtilis mutant 2 and the wild type did not result in a growth performance different from the negative and positive controls. In conclusion, results obtained with the mutant strains of Bacillus subtilis were not better than results obtained with the wild-type strain, and for both strains, the results were not different than the negative control.

The structure of a bottlenose dolphin society is coupled to a unique foraging cooperation with artisanal fishermen.

Diverse and localized foraging behaviours have been reported in isolated populations of many animal species around the world. In Laguna, southern Brazil, a subset of resident bottlenose dolphins (Tursiops truncatus) uses a foraging tactic involving cooperative interactions with local, beach-casting fishermen. We used individual photo-identification data to assess whether cooperative and non-cooperative dolphins were socially segregated. The social structure of the population was found to be a fission-fusion system with few non-random associations, typical for this species. However, association values were greater among cooperative dolphins than among non-cooperative dolphins or between dolphins from different foraging classes. Furthermore, the dolphin social network was divided into three modules, clustering individuals that shared or lacked the cooperative foraging tactic. Space-use patterns were not sufficient to explain this partitioning, indicating a behavioural factor. The segregation of dolphins using different foraging tactics could result from foraging behaviour driving social structure, while the closer association between dolphins engaged in the cooperation could facilitate the transmission and learning of this behavioural trait from conspecifics. This unique case of a dolphin-human interaction represents a valuable opportunity to explore hypotheses on the role of social learning in wild cetaceans.

CpG island methylator phenotype (CIMP) of colorectal cancer is best characterised by quantitative DNA methylation analysis and prospective cohort studies.

BACKGROUND: The concept of CpG island methylator phenotype (CIMP) is not universally accepted. Even if specific clinicopathological features have been associated with CIMP, investigators often failed to demonstrate a bimodal distribution of the number of methylated markers, which would suggest CIMP as a distinct subtype of colorectal cancer. Previous studies primarily used methylation specific polymerase chain reaction which might detect biologically insignificant low levels of methylation. AIM: To demonstrate a distinct genetic profile of CIMP colorectal cancer using quantitative DNA methylation analysis that can distinguish high from low levels of DNA methylation. MATERIALS AND METHODS: We developed quantitative real time polymerase chain reaction (MethyLight) assays and measured DNA methylation (percentage of methylated reference) of five carefully selected loci (promoters of CACNA1G, CDKN2A (p16), CRABP1, MLH1, and NEUROG1) in 460 colorectal cancers from large prospective cohorts. RESULTS: There was a clear bimodal distribution of 80 microsatellite instability-high (MSI-H) tumours according to the number of methylated promoters, with no tumours showing 3/5 methylated loci. Thus we defined CIMP as having >or=4/5 methylated loci, and 17% (78) of the 460 tumours were classified as CIMP. CIMP was significantly associated with female sex, MSI, BRAF mutations, and wild-type KRAS. Both CIMP MSI-H tumours and CIMP microsatellite stable (MSS) tumours showed much higher frequencies of BRAF mutations (63% and 54%) than non-CIMP counterparts (non-CIMP MSI-H (0%, p<10(-5)) and non-CIMP MSS tumours (6.6%, p<10(-4)), respectively). CONCLUSION: CIMP is best characterised by quantitative DNA methylation analysis. CIMP is a distinct epigenotype of colorectal cancer and may be less frequent than previously reported.

An evaluation of hybrid methods for matching biomedical terminologies: mapping the gene ontology to the UMLS.

Integration of disparate biomedical terminologies is becoming increasingly important as links between biological science and clinical medicine grow. Mapping concepts in the Gene Ontology (GO) to the UMLS may help further this integration and allow for more efficient information exchange among researchers. Using a gold standard of GO term--UMLS concept mappings provided by the NCI, we examined the performance of various published and combined mapping techniques, in order to maximize precision and recall. We found that for the previously published techniques precision varied between (0.61-0.95), and recall varied from (0.65-0.90), whereas for the hybrid techniques, precision varied between (0.66-0.97), and recall from (0.59-0.93). Our study reveals the benefits of using mapping techniques that incorporate domain knowledge, and provides a basis for future approaches to mapping between distinct biomedical vocabularies.