A comprehensive molecular analysis of bovine coronavirus strains isolated from Brazil and comparison of a wild-type and cell culture-adapted strain associated with respiratory disease.

Bovine coronavirus (BCoV) has dual tropisms that can trigger enteric and respiratory diseases in cattle. Despite its global distribution, BCoV field strains from Brazil remain underexplored in studies investigating the virus's worldwide circulation. Another research gap involves the comparative analysis of S protein sequences in BCoV isolates from passages in cell lines versus direct sequencing from clinical samples. Therefore, one of the objectives of our study was to conduct a comprehensive phylogenetic analysis of BCoV strains identified from Brazil, including a respiratory strain obtained during this study, comparing them with global and ancestral BCoV strains. Additionally, we performed a comparative analysis between wild-type BCoV directly sequenced from the clinical sample (nasal secretion) and the cell culture-adapted strain, utilizing the Sanger method. The field strain and multiple cell passage in cell culture (HRT-18) adapted BCoV strain (BOV19 NS) detected in this study were characterized through molecular and phylogenetic analyses based on partial fragments of 1,448 nt covering the hypervariable region of the S gene. The analyses have demonstrated that different BCoV strains circulating in Brazil, and possibly Brazilian variants, constitute a new genotype (putative G15 genotype). Compared with the ancestral prototype (Mebus strain) of BCoV, 33 nt substitutions were identified of which 15 resulted in non-synonymous mutations (nine transitions and six transversions). Now, compared with the wild-type strain was identified only one nt substitution in nt 2,428 from the seventh passage onwards, which resulted in transversion, neutral-neutral charge, and one substitution of asparagine for tyrosine at aa residue 810 (N810Y).

High genetic merit dairy heifers grazing low quality forage had similar weight gain and urinary nitrogen excretion to those of low genetic merit heifers.

Climate variability and increasing drought events have become significant concerns in recent years. However, there is limited published research on body weight (BW) change of dairy heifers with different genetic merit when grazing on drought impacted pastures in southern Australia. Achieving target body weight (BW) is vital for dairy heifers, especially during critical stages like mating and calving. This study aimed to assess dry matter (DM) intake, BW change, urinary nitrogen excretion, and grazing behaviours of high vs. low genetic dairy heifers grazing pasture during a 43-day experimental period in a drought season. Forty-eight Holstein Friesian heifers grazed on ryegrass-dominant pasture and were divided into two groups based on their high and low Balanced Performance Index (HBPI and LBPI, respectively). Each group was further stratified into six plots, with similar BW, resulting in four heifers per replication group. Data from the five measurement days were averaged for individual cows to analyse the dry matter intake, nitrogen intake and nitrogen excretion. The statistical model included the treatment effect of BPI (H and L) and means were analysed using ANOVA. The pasture quality was poor, with metabolizable energy 9.3 MJ/Kg DM and crude protein 5.9% on a DM basis. Nitrogen intake and urinary nitrogen excretion were significantly higher (p < 0.05) in HBPI compared to the LBPI. However, despite these differences, the study did not find any advantages of having HBPI heifer grazing on low quality forage in terms of BW performance.

Effects of vitamin E and selenium administration on transportation stress in pregnant dairy heifers.

We investigated the effects of road transportation and administration of the vitamin E and selenium (ESe) on circulating cortisol, haptoglobin, blood metabolites, oxidative biomarkers, white blood cell profiles, and behaviors in pregnant dairy heifers. Forty pregnant Holstein heifers were randomly assigned to one of 4 treatments: no transportation and no ESe administration, no transportation and ESe administration, transportation and no administration, and transportation and ESe administration. The ESe (70 IU/kg dry matter feed of dl-α-tocopheryl acetate and 0.3 mg/kg dry matter feed of sodium selenite) was orally delivered once a day from 7 d before transportation to 3 d after transportation. The heifers were transported in trucks designed for cattle transportation. Blood was collected 1 h before transportation, immediately after transportation (IAT), and at 6, 24, and 48 h after transportation. Behaviors were recorded using a video camera for 2 consecutive days after transportation. Transported/non-ESe-administered heifers had greater cortisol at IAT, haptoglobin at 6 and 24 h after transportation, total oxidative status at 6 h after transportation, and nonesterified fatty acid levels, white blood cell numbers, and neutrophil percentages at IAT and 6 h after transportation in the blood than nontransported heifers. Transported/non-ESe-administered heifers had lower total antioxidative status levels at 48 h after transportation and lymphocyte percentages at IAT and 6 h after transportation than nontransported heifers. Lying time was shorter in transported heifers than nontransported/non-ESe-administered heifers. Transported/ESe-administered heifers had lower cortisol, total oxidative status, nonesterified fatty acid levels at IAT, and haptoglobin concentrations at 6 and 24 h after transportation than transported/non-ESe-administered heifers. Transported/ESe-administered heifers had greater total antioxidative status levels at 48 h after transportation than transported/non-ESe-administered heifers. No ESe administration effects were observed for white blood cell number and neutrophil and lymphocyte percentages and lying time. In conclusion, road transportation caused temporary oxidative stress. Administrating ESe partially alleviated the stress, suggesting that ESe administration could be a viable strategy to reduce stress in transported pregnant heifers, providing a novel role of vitamin E and selenium for improving animal welfare.

Limit feeding total mixed rations exacerbates intersucking in year-old dairy heifers.

Limit feeding is a practice that is used to improve feed efficiency and control growth in dairy heifers, but also has negative consequences associated with hunger and restriction of feeding behavior. One such consequence could be intersucking (i.e., the licking or sucking of another animal's teats or udder). This behavior is reported to varying degrees in heifers, and thus, our first objective was to evaluate whether intersucking increased in response to short-term (48 h) feed restriction. Intersucking interventions, such as nose rings, are often applied to specific animals, and so our second objective was to describe intersucking performance in individual heifers, including those that represent outliers in this population. We studied 42 heifers (9 Jerseys, 33 Holsteins) aged 12.8 ± 1.1 mo (mean ± SD). They were housed in 21 pairs, the experimental unit for our first objective. We used a switchback design with 3 periods of 2 d each where heifers were fed ad libitum total mixed ration in the first and third periods (Baseline and Return, respectively), and limit fed to 50% of ad libitum intake in the second period (Restriction). We scored time spent intersucking continuously and eating at 5-min intervals from video recordings from 0800 to 2000 h on 4 d (baseline d 2, restriction d 2, return d 1, and return d 2). Heifer pairs spent less time eating (16 ± 0.4% mean percentage of 12-h observations ± SE), more time intersucking (27 ± 7 s/12 h), and engaged in longer bouts of intersucking (23 ± 5 s/bout) on Restriction d 2 compared with all other days (eating: 34.7 ± 1.1%, 44.2 ± 0.9%, 35.8 ± 1.1% of 12-h; intersucking time: 7 ± 2, 7 ± 3, 10 ± 5 s/12 h; intersucking bout length: 3 ± 1, 2 ± 1, 3 ± 1 s/bout; Baseline d 2, Return d 1, Return d 2, respectively). There was no difference in the number of bouts of intersucking across days (1.3 ± 0.2 bouts/12 h). The drop in eating during feed restriction was followed by a rebound on Return d 1 before returning to baseline levels. Time spent intersucking did not differ among ad libitum periods. Overall, 90% of the heifers performed intersucking on at least 1 of the 4 d, and did this for 1 to 127 s/12 h (range) in 1 to 13 bouts/12 h. Of the heifers that intersucked, 55% did this at extreme levels relative to the rest of the experimental population (outliers). Solid feed restriction exacerbated intersucking in year-old heifers, but this behavior was widespread.

The effect of progesterone concentrations during superovulation of Holstein heifers in a randomized trial.

The aim of this study was to evaluate the effect of different progesterone (P4) concentrations during the follicular growth on the intensity of estrous expression, ovarian response to the superovulatory treatment, and embryo production and quality in superovulated heifers. A total of 63 Holstein heifers were randomly assigned into 2 experimental groups: Low P4 (n = 31) and High P4 (n = 32). Animals received a pre-synchronization protocol followed by a protocol of superovulation that included the allocated P4 treatment. Activity was monitored continuously by an automated activity monitor, and estrus characteristics (maximum intensity and duration) were recorded. Embryo collection was performed 7 d post artificial insemination (AI). Embryos were counted and graded from good or excellent (1) to degenerated (4). The outcomes of interest were: number and diameter of follicles at the time of AI, ovulation success (confirmed 7 d post-AI), time to estrus event, maximum intensity and duration of estrus, number and quality of embryos. Data were analyzed according to the type of outcome variable using logistic, linear, or Poisson regression models. A total of 105 embryos (High P4: n = 42; Low P4: n = 63) were graded for quality. Different P4 levels did not affect the maximum intensity (High P4 = 497.8 ± 23.9%; Low P4 = 542.2 ± 23.5%) or the duration (High P4 = 13.5 ± 1.5 h; Low P4 = 14.3 ± 1.4 h) of estrus. Heifers in the High P4 treatment had greater number of follicles at time of AI (High P4 = 16.6 ± 1.6 follicles; Low P4 = 13.9 ± 1.2 follicles), but with smaller diameter (High P4 = 11.3 ± 0.1 mm; Low P4 = 12.0 ± 0.1 mm) compared with Low P4. High P4 heifers tended to have better embryo quality compared with Low P4 heifers (odds ratio = 1.98; 95% CI = 0.90-4.35). High P4 heifers had less embryos than Low P4 heifers, but this was modified by the CIDR (intravaginal implant of P4) removal to estrus interval (interval 0-21 h: mean ratio = 1.15, 95% CI = 0.42-1.87; interval 22-46 h: mean ratio = 0.58, 95% CI = 0.27-0.96). Although estrous expression was not associated with embryo quality, as the duration and the maximum intensity of estrous expression increased, the number of embryos recovered 7 d post-AI increased (duration: mean ratio = 1.04; 95% CI = 1.03-1.05; maximum intensity: mean ratio = 1.50; 95% CI = 1.42-1.58). In conclusion, P4 during the follicular growth, and intensity of estrus, are playing a role in regulating the quality and the number of embryos produced by superovulated heifers. This study was supported by contributions from Resilient Dairy Genome Project and the Natural Sciences and Engineering Research Council.

Animal- and herd-level factors associated with onset of puberty in grazing dairy heifers.

AIMS: To explore animal- and herd-level risk factors influencing age at puberty in predominantly Holstein-Friesian dairy heifers managed in seasonal, pasture-based systems. METHODS: Heifers born in spring 2018 (n = 5,010) from 54 commercial dairy herds in New Zealand were visited on three occasions when the mean heifer age, within herd, was 10 (visit 1; V1), 11 (V2) and 12 (V3) months old. Blood samples were collected on each visit and liveweight, stature and anogenital distance (AGD) were measured at V2. Heifers were defined as having reached puberty at the first visit where blood progesterone was elevated (≥ 1 ng/mL). Animal-level response variables included pubertal status by V1, V2 and V3, and age at puberty (or age at V3 plus 31 days for those that had not attained puberty by V3). To explore herd-level management factors, farmers answered a questionnaire relating to animal location, land type, health, feeding, and management between weaning and mating. A partial least squares regression was undertaken to identify herd-level factors associated with the greatest influence on puberty rate within herd. RESULTS: The mean age at puberty was 352 (SD 34.9) days. Heavier animals at a greater proportion of expected mature liveweight based on their breeding value for liveweight, or animals with a higher breed proportion of Jersey and lower breed proportion of Holstein, were associated with earlier puberty. Herd puberty rates varied widely among enrolled herds, and averaged 20%, 39% and 56% by V1, V2 and V3, respectively. Liveweight, followed by breed and land type, had the greatest influence on the herd puberty rate. Heifer herds with a greater mean liveweight (absolute and proportion of expected mature weight) or greater Jersey proportion had more animals that reached puberty at any visit, whereas herds located on steep land or with greater Holstein breed proportions had lower puberty rates. Management-related factors such as vaccinations, provision of feed supplements, and weighing frequency were also herd-level risk factors of puberty but had less influence. CONCLUSIONS AND CLINICAL RELEVANCE: This study highlights the importance of having well-grown heifers for increasing the chances of earlier puberty onset and the effect of breed and youngstock management to achieve growth targets. These outcomes have important implications for the optimal management of heifers to achieve puberty before their maiden breeding and for the timing of measurements to potentially incorporate a puberty trait in genetic evaluations.

The anogenital ratio as an indicator of reproductive performance in dairy heifers.

The anogenital distance (AGD) is considered a marker for prenatal androgen exposure and fertility in multiple species including humans. In dairy cattle, it is described as the length between the center of the anus and the clitoral base (AGDc). However, in other species, the distance from the center of the anus to the dorsal commissure of the vulva (AGDv) is also considered to be a predictor for fertility traits, as well as the anogenital ratio (AGR, defined as [AGDv/AGDc]*100). The primary aim of the present study was to assess whether AGDv and AGR can be used as an indicator for reproductive performance in dairy heifers. Additionally, the relation between AGDv and AGDc and the correlation with other body measurements were explored. Data of 656 Holstein Friesian heifers at an age of 13.5 ± 1.08 months were analyzed. Respective means of 62.9 ± 8.20 mm (AGDv) and 107.6 ± 9.27 mm (AGDc) were recorded. The mean AGR ratio was calculated as 58.6 ± 6.75%, varying from 37.3 to 79.6%. The age of the heifers was not associated with any of the AGD measurements nor the ratio. Except for a very low correlation between heart girth and AGDc (r = 0.09, P < 0.05), both AGDs were largely uncorrelated with other body measurements. Linear regression models revealed that AGDc was not associated with any of the recorded fertility parameters. However, results revealed a negative association between AGDv and AGR and reproductive performance: heifers with a short AGDv and small AGR were younger at first AI (P ≤ 0.003) and at conception (P = 0.004). Based on ROC curve analyses, AGDv was the best indicator for pregnancy to first AI, with a threshold estimated at 65.3 mm. The pregnancy rate at first AI was 72.4% in heifers with a short AGDv (<65.3 mm, n = 413) compared to 61.7% in heifers with a long AGDv (≥65.3 mm, n = 243). Hence, short-AGDv heifers had 63% higher odds to conceive at first AI compared to their long-AGDv counterparts (P = 0.004). Additionally, an AGR threshold of 59,6% was determined: heifers with a small AGR (<59.6%) had 44% higher odds to be pregnant at first AI compared to heifers with an AGR ≥59.6%. Results of the present study suggest to consider AGDv and AGR as potential indicators for reproductive performance in dairy heifers. The latter implies that it is relevant to measure both AGDc and AGDv in future studies. The absence of correlation between body- and AGD-measurements furthermore suggests that AGD sizes are rather pre-than postnatally determined.

Ultrasound characterization of mammary gland development in heifer calves fed at two different levels until weaning.

Ultrasound technologies allow for a non-invasive assessment of mammary gland (MG) development, the differentiation between the tissue types of the MG, and the evaluation of changes in its composition. This study aimed to work out a detailed description of the different stages of MG development that are visually discernible by ultrasonography for providing a template to classify the different structures. With this basis, the qualitative categorization of the developmental stage, as well as further quantitative assessments via pixel densities in the structures of interest, should be facilitated. Ultrasonic images were acquired from all four quarters of 37 German Holstein heifer calves fed either at a high feeding level of milk replacer (MR; 14% solids) at 10 L/day (1.4 kg MR/day; HI, n = 18) or at a restrictive low level of 5.7 L/day (0.8 kg MR/day; RES, n = 19) until linear weaning from week 13 to 14 of life. Ultrasound MG scans were performed first in week 3 of life, fortnightly from week 8-16, and in week 20 of life, in standing position, of each quarter, using a B-mode ultrasound device equipped with a linear probe (18 MHz). The developmental stages of the mammary gland parenchyma (PAR), visible in ultrasound images, obtained over 20 weeks of life, were categorized, described, and drawn by hand. On this basis, a template for classifying the visible categories of mammary PAR development and its surrounding tissue (SURR), and for measuring their pixel brightness was created thus providing an ultrasonographic atlas of the developing bovine MG, describing 11 categories. The ultrasound images were further classified by PAR structure, and pixel brightness was measured in PAR and SURR by using ImageJ Fiji. The difference in pixel brightness between PAR and SURR, the delta (Δ) pixel value was calculated. With increasing age, higher atlas categories of PAR developmental stages were shown. Pixel values, i.e. the brightness of PAR, its SURR, and Δ pixel value changed with age but were neither affected by the feeding group nor by a group × time interaction. With progressing PAR development, its pixel brightness increased from week 10 to 20 of life, i.e., the PAR became more hyperechoic since it spread and grew into its SURR. The atlas can serve as a template for the categorization and qualitative assessment of MG structures and for the quantitative assessment of PAR's development by measuring pixel brightness. With our study, we could show the structural development in PAR as well as in SURR in MG simultaneously in early life and confirm the spreading of PAR into its SURR by ultrasound scanning.

Evaluation of a Novel Infrared Thermography Projection to Assess Udder Health in Primigravid Dairy Heifers.

Heifer mastitis in early lactation impacts negatively on animal welfare, milk production and longevity. A major challenge for the prevention and control of mastitis in dairy heifers is to establish when intramammary infection occurs because pre-partum secretum sampling is risky. We evaluated a ventrodorsal projection to capture thermal images of the entire udder of primigravid and compared results against caudocranial projection, which is used in lactating cattle. Based on the analysis of 119 heifers and images taken at 2 months and 2 weeks pre-partum, a very strong positive correlation (r = 0.91 and r = 0.96, respectively) was shown between caudocranial and ventrodorsal projections of hind quarters. Quarter maximum gradient temperatures were consistently greater on ventrodorsal projection than on caudocranial projection, and less variable than minimum gradient temperatures. The collection of ventrodorsal images is a simple one-step method involving the imaging of the entire udder in a manner safe for both the cattle and handlers. Together, these results demonstrate that a single projection can be used to scan the entire udder of primigravid dairy heifers in commercial farm conditions, with the potential to implement this as a routine method for the early detection of intramammary infection based on udder surface temperature.

Modelling growth in dairy heifers based on linear body measurements (withers height) using non-linear functions.

This research communication reports a study to model the growth curves for withers height (WH) and body weight (BW) to withers height ratio (BW:WH) using monthly records (from 1 to 24 months of age) for three breeds of dairy heifer (Holstein, Jersey and Brown Swiss). The data sets used were those reported by the Dairy Heifer Evaluation Project of Penn State Extension (USA) between 1991 and 1992. Four growth functions (monomolecular and Michaelis-Menten, both with diminishing returns behaviour, and Schumacher and López, both with asymptotic sigmoidal behaviour) were fitted using the non-linear regression procedure of the SigmaPlot software and the parameters estimated. The models were judged for goodness of fit using adjusted coefficient of determination ($R_{{\rm adj}}^2 $), root mean square error (RMSE), Akaike's information criterion (AIC) and Bayesian information criterion (BIC). Assessing the goodness of fit by $R_{{\rm adj}}^2 $ (>0.99 in all cases) reveals the generally appropriate fit of the models to the data. The non-sigmoidal functions (i.e. Michaelis-Menten and monomolecular) provided the best fits giving the lowest values of RMSE, AIC and BIC. Based on the chosen statistical criteria, the Schumacher and López equations provided acceptable fits to the WH and BW:WH growth curves, but showed points of inflexion at times before birth, indicating that these growth curves are not sigmoidal. In conclusion, evaluation of the different non-linear growth functions used in this study indicated their potential for modelling growth patterns in dairy heifers.

Effect of diet energy level and genomic residual feed intake on bred Holstein dairy heifer growth and feed efficiency.

The objective of this study was to determine growth, feed intake, and feed efficiency of postbred dairy heifers with different genomic residual feed intake (RFI) predicted as a lactating cow when offered diets differing in energy density. Postbred Holstein heifers (n = 128, ages 14-20 mo) were blocked by initial weight (high, medium-high, medium-low, and low) with 32 heifers per block. Each weight block was sorted by RFI (high or low) to obtain 2 pens of heifers with high and low genomically predicted RFI within each block (8 heifers per pen). Low RFI heifers were expected to have greater feed efficiency than high RFI heifers. Dietary treatments consisted of a higher energy control diet based on corn silage and alfalfa haylage [HE; 62.7% total digestible nutrients, 11.8% crude protein, and 45.6% neutral detergent fiber; dry matter (DM) basis], and a lower energy diet diluted with straw (LE; 57.0% total digestible nutrients, 11.7% crude protein, and 50.1% neutral detergent fiber; DM basis). Each pen within a block was randomly allocated a diet treatment to obtain a 2 × 2 factorial arrangement (2 RFI levels and 2 dietary energy levels). Diets were offered in a 120-d trial. Dry matter intake by heifers was affected by diet (11.0 vs. 10.0 kg/d for HE and LE, respectively) but not by RFI or the interaction of RFI and diet. Daily gain was affected by the interaction of RFI and diet, with low RFI heifers gaining more than high RFI heifers when fed LE (0.94 vs. 0.85 kg/d for low and high RFI, respectively), but no difference for RFI groups when fed HE (1.16 vs. 1.19 kg/d for low and high RFI, respectively). Respective feed efficiencies were improved for low RFI compared with high RFI heifers when fed LE (10.6 vs. 11.8 kg of feed DM/kg of gain), but no effect of RFI was found when fed HE (9.4 vs. 9.5 kg of DM/kg of gain for high and low RFI, respectively). No effect of RFI or diet on first-lactation performance through 150 DIM was observed. Based on these results, the feed efficiency of heifers having different genomic RFI may be dependent on diet energy level, whereby low RFI heifers utilized the LE diet more efficiently. The higher fiber straw (LE) diet controlled intake and maintained more desirable heifer weight gains. This suggests that selection for improved RFI in lactating cows may improve feed efficiency in growing heifers when fed to meet growth goals of 0.9 to 1.0 kg of gain/d.

Effects of stress after road transportation and oral administration of chromium and meloxicam on plasma cortisol concentrations and behavior in dairy calves.

OBJECTIVE: This study was performed to determine the effects of stress after road transportation and oral administration of chromium and meloxicam on growth performance, plasma cortisol, serum metabolites, and behavior in dairy calves. METHODS: A total of 50 Holstein heifers (average body weight [BW]: 172±4.19 kg; average age: 5.53±0.12 months) were randomly assigned to five groups including NL (not transported + D-lactose; 1 mg/kg BW), TL (transported + D-lactose; 1 mg/kg BW), TC (transported + chromium; 0.5 mg/kg dry matter [DM] feed), TM (transported + meloxicam; 1 mg/kg BW), and TMC (transported + combination of meloxicam and chromium; 1 mg/kg BW and 0.5 mg/kg DM, respectively). Doses of D-lactose monohydrate, meloxicam, and chromium were prepared for oral administration by suspension in 15 mL of water in a 20-mL dosing syringe. Blood was collected before transportation, immediately after 120 km of transportation (IAT), and at 6, 24, and 48 h after transportation. RESULTS: Neither transportation nor administration of meloxicam and/or chromium affected (p = 0.99) average daily gain and feed intake. Plasma cortisol concentrations in the NL group (average 0.13 and 0.18 nmol/L, respectively) were lower (p<0.001) compared to the TL group (average 0.39 and 0.61 nmol/L, respectively) at IAT and 48 h after transportation. At 48 h after transportation, cortisol concentrations were lower (p<0.05) in the TC group (average 0.22 nmol/L) than in the TL group (average 0.61 nmol/L), and TC calves had similar cortisol concentrations to NL calves. Lying duration (min/d) was shorter (p<0.05) in the TL group than in the NL group at 2 d after transportation. Lying duration was longer (p<0.05) for the TC and TMC groups than for the TL group at 2 d after transportation. CONCLUSION: Transportation increased cortisol concentrations and affected lying behavior, while chromium administration reduced cortisol concentrations and changed lying behavior. Thus, chromium administration before transportation may be a viable strategy to alleviate stress elicited by road transportation.

Comparison of reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen based on expression of estrus, pregnancy outcomes, and cost per pregnancy.

Our objective was to evaluate reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen. In experiment 1, nulliparous Holstein heifers (n = 462) were submitted to a 5-d progesterone-releasing intravaginal device (PRID)-Synch protocol [d 0, GnRH + PRID; d 5, PGF2α - PRID; d 6, PGF2α; d 8, GnRH + TAI] and were randomly assigned for PRID removal on d 5 or 6 of the protocol followed by timed artificial insemination (TAI) with conventional semen. Delaying PRID removal decreased early expression of estrus before scheduled TAI (0.9 vs. 12.2%), and pregnancies per AI (P/AI) did not differ between treatments. In experiment 2, nulliparous Holstein heifers (n = 736) from 3 commercial farms were randomized within farm to 1 of 3 treatments for first AI with sexed semen: (1) CIDR5 [d -6, GnRH + controlled internal drug release (CIDR); d -1, PGF2α - CIDR; d 0, PGF2α; d 2, GnRH + TAI]; (2) CIDR6 (d -6, GnRH + CIDR; d -1, PGF2α; d 0, PGF2α - CIDR; d 2, GnRH + TAI); and (3) EDAI (PGF2α on d 0 followed by once-daily estrous detection and AI). Delaying CIDR removal decreased early expression of estrus before scheduled TAI (0.004 vs. 27.8%); however, CIDR5 heifers tended to have more P/AI at 35 (53 vs. 45 vs. 46%) and 64 (52 vs. 45 vs. 45%) days after AI than CIDR6 and EDAI heifers, respectively. Overall, CIDR5 and CIDR6 heifers had fewer days to first AI and pregnancy than EDAI heifers which resulted in less feed costs than EDAI heifers due to fewer days on feed until pregnancy. Despite greater hormonal treatment costs for CIDR5 heifers, costs per pregnancy were $16.66 less for CIDR5 than for EDAI heifers. In conclusion, delaying PRID removal by 24 h within a 5-d PRID-Synch protocol in experiment 1 suppressed early expression of estrus before TAI, and P/AI for heifers inseminated with conventional semen did not differ between treatments. By contrast, although delaying CIDR removal by 24 h within a 5-CIDR-Synch protocol in experiment 2 suppressed early expression of estrus before TAI, delaying CIDR removal by 24 h tended to decrease P/AI for heifers inseminated with sexed semen. Further, submission of heifers to a 5-d CIDR-Synch protocol for first AI tended to increase P/AI and decrease the cost per pregnancy compared with EDAI heifers.

Effect of One-Day Delaying CIDR Administration in 5-Day Cosynch Protocol in Dairy Heifers.

Effect of one-day delaying progesterone administration at the beginning of 5-day Cosynch protocol was investigated in Holstein heifers for the first artificial insemination (AI). Heifers received a synchronized ovulation and timed AI (TAI) with CIDR inserted on day (d) 0 (CIDR-5; n = 206) or d 1 (CIDR-4; n = 192). In both group, GnRH was administered on d 0 followed by a single PGF2α injection and CIDR removal five days later from GnRH. On d 8, TAI and GnRH administration were concurrently conducted. Heifers detected in estrus up to 24 h prior to TAI were inseminated without GnRH administration. Rates of ovulation, accessory CL formation and new dominant follicle development following initial GnRH injection did not differ between groups. P/AI did not differ between CIDR-4 (44.3%, [85/192]) and CIDR-5 (51.9%, [107/206]) groups, respectively. Pregnancy per AI (P/AI) was significantly (p < 0.01) declined as heifers' age (12-13, 14, 15, 16 and17-21 months) proceeded in CIDR-4 group (55.6%, 52.1%, 37.9%, 35.7%, 32.4%) compared to those in CIDR-5 group (60.0%, 50.0%, 53.9%, 51.5%, 46.2%) respectively. In conclusion, there is no benefit for delaying CIDR administration in 5-day Cosynch protocol in dairy heifers. However, higher P/AI in CIDR-5 group in older heifers can be considered for reproductive management.

Effect of PGF2α treatments during early corpus luteum development on circulating progesterone concentrations and ovulation in breeding-age Holstein heifers.

The objective of this study was to test the effect of low circulating concentrations of progesterone (P4) on pre-ovulatory follicle development in heifers as part of an overarching objective to develop a model to understand this phenomenon in dairy cattle without the confounding factors of lactation. Holstein heifers between 12 and 13 mo of age were pre-synchronized to ensure all heifers were on d 6 of the estrous cycle at the start of the Ovsynch program. Only heifers with CL regression and ovulation to the following pre-treatment strategy were used in the study: 0.5 mg cloprostenol (PGF2α), 2 d later, 0.1 mg GnRH, 6 d later, GnRH (G1; 1st GnRH of Ovsynch). Heifers (n = 159) responding to pre-treatment were randomly assigned to 4 groups and completed the Ovsynch program: high P4 control (HPC), low P4 control (LPC; PGF2α 24 h after G1), PG2 (PGF2α 24 and 48 h after G1) and PG3 (PGF2α 24, 48, and 96 h after G1). Only heifers that had ovulation to G1 remained in the study. Blood samples were collected in all heifers on d 7 (n = 157) and in a subset of heifers on d 1, 2, 3, 4 (n = 82) after G1 to measure serum P4. Pre-ovulatory follicle size at G1 (13.0 ± 0.1 mm; P = 0.53) and mean serum P4 24 h after G1 (d 1; 3.62 ± 0.11 ng/mL; P = 0.46) did not differ among treatments. HPC heifers had greater (P < 0.001) mean serum P4 compared to LPC, PG2 and PG3 on d 2, 3, 4, and 7. On d 2, 3 and 4, mean serum P4 of LPC, PG2 and PG3 heifers did not differ (P > 0.10). On d 7, LPC heifers had greater (P < 0.001) serum P4 compared to PG2 and PG3 heifers. Mean ± SEM serum P4 on d 7 after G1 was 8.43 ± 0.39, 2.55 ± 0.36, 1.58 ± 0.20, and 1.21 ± 0.15 ng/mL for HPC, LPC, PG2, and PG3, respectively. Percentage of heifers with P4 < 0.50 ng/mL on d 7 was greater (P < 0.05) for LPC, PG2 and PG3 (27, 32 and 26%, respectively) compared to HPC (0%). A greater (P < 0.05) proportion of heifers ovulated before G2 in the LPC, PG2 and PG3 than in the HPC. For heifers that ovulated after G2, low serum concentrations of P4 in LPC, PG2 and PG3 induced double ovulations in 6/97 heifers after the final GnRH of Ovsynch compared to 0/33 in HPC. In summary, PGF2α treatments during early CL development reduced circulating P4 concentrations 7 d after G1 compared with both HPC and LPC. However, it did not effectively control CL and follicle function to be utilized as a model to test high vs. low serum P4 on fertility parameters in Holstein heifers.

The effects of organic grass and grass-birdsfoot trefoil pastures on Jersey heifer development: Herbage characteristics affecting intake.

Low dietary energy and decreased intake of herbage have been attributed to the reduced performance of grazing dairy cattle. We hypothesized that grasses with inherently greater energy would interact in a complementary way with condensed tannins (CT) in birdsfoot trefoil to increase herbage intake by grazing dairy heifers. Eight pasture treatments comprising high-sugar perennial ryegrass (Lolium perenne L.), orchardgrass (Dactylis glomerata L.), meadow bromegrass (Bromus riparius Rehmann), and tall fescue [Schendonorus arundinaceus (Schreb.) Dumort] were established in Lewiston, Utah as monocultures and binary mixtures with birdsfoot trefoil (Lotus corniculatus L.; BFT). Pasture treatments were rotationally stocked by Jersey heifers for 105 d in 2017 and 2018, and herbage samples were collected pre- and postgrazing for each 7-d grazing period and analyzed for herbage mass, nutritive value, and apparent herbage intake. We observed differences among pasture treatments in herbage quantity and nutritive value, as well as differences in herbage intake by grazing Jersey heifers. On average, grass-BFT mixtures had greater herbage intake than grass monocultures, and every grass-BFT treatment individually had greater herbage intake than their respective grass monocultures. Using multivariate analyses, we determined that approximately 50% of the variation in herbage intake was due to nutritive and physical herbage characteristics, with the most explanatory being characteristics related to fiber and energy, followed by those related to the percent of BFT in the herbage. Grass monocultures exhibited a range of inherent dietary energy, but there was indication that an imbalance of energy to crude protein (e.g., protein deficient) reduced intake of grass monocultures. Moreover, there was some evidence of a complementary effect between increased dietary energy and CT; however, low CT levels made it impossible to determine the effect of CT on herbage intake per se. This study confirmed that chemical and physical characteristics inherent to different pasture species have a large effect on herbage intake by grazing cattle. Pastures planted to binary mixtures of nutritious grasses and birdsfoot trefoil increase herbage intake of temperate pastures by grazing Jersey heifers.

Characteristics of the Oxidative Status in Dairy Calves Fed at Different Milk Replacer Levels and Weaned at 14 Weeks of Age.

A paradigm shift in the way of rearing heifer calves from restricted feeding and early weaning towards greater feed allowances and later weaning ages is ongoing. We aimed at characterizing the oxidative status in Holstein heifer calves fed with milk replacer (MR) at either a restrictive (RES) or a high (HIGH) level for 14 weeks. We compared two groups: HIGH (10 L MR/d, n = 18) and RES (5.7 L/d, n = 19) from day five until week 14 of life. In blood samples collected at birth, and then fortnightly from week 8-16, and in week 20, the antioxidative capacity measured as ferric reducing ability of plasma (FRAP), oxidative damage of lipids measured as thiobarbituric acid reactive substances (TBARS) and oxidative damage of proteins measured as advanced oxidation products of proteins (AOPP), free radicals measured as reactive oxidative metabolites (dROM), and the glutathione peroxidase (GSH-Px) activity, as well as leptin, adiponectin and haptoglobin were assessed. The time course of these variables during the first 20 weeks of life showed characteristic patterns; group differences were limited to adiponectin, AOPP, and FRAP. RES calves had lower growth rates, showed signs of hunger, but did not differ from HIGH in their intake of solid starter feed and in health status. This work characterizes the changes in oxidative status of dairy calves with increasing age and confirms the benefits of a high feeding plane with regard to welfare and development.

Changes in the soft-tissue thickness of the claw sole in Holstein heifers around calving.

Claw horn disruption lesions are a common cause of lameness in dairy cattle. It is commonly agreed that they develop due to excessive pressure from the distal phalanx on the horn-producing tissue in the sole. The distal phalanx is supported by the suspensory apparatus, a large proportion of which is made up of connective tissue. It has been hypothesized that increased laxity of the connective tissue due to hormonal changes around calving may lead to decreased support of the distal phalanx, resulting in a sinking of the bone and increased pressure on the horn-producing tissue. In this longitudinal study, our first objective was to determine whether the thickness of the soft tissue related to the claw sole (the combined depth of the digital cushion and corium; soft-tissue thickness, STT) in the weight-bearing hind claws of live heifers changed around calving (the period from approximately 3 wk before calving to 2 wk after calving). Our second objective was to evaluate the relationship between the body condition score (BCS) of recruited heifers and the STT estimates we obtained. We measured STT (defined as the distance between the distal phalanx and the sole horn) in the apex and the posterior part of the sole of the hind claws of 34 Holstein heifers 4 times over 3 to 4 wk around calving. We determined STT by ultrasonographic measurements through the sole horn on weight-bearing claws. The overall mean STT at site 1 was 3.3 mm [minimum-maximum (min-max): 1.8 to 5.2 mm], and the mean at site 2 was 3.7 mm (min-max: 2.2 to 5.8). The overall mean STT was thickest in the first examination before calving [site 1: 3.4 mm (min-max: 1.9 to 4.7); site 2: 4.1 mm (min-max: 2.7 to 5.8)] and thinnest at site 1 in the second examination after calving (3.2 mm, min-max: 1.8 to 5.2) and at site 2 at the first examination after calving (3.5 mm, min-max: 2.2 to 5.0). We observed an average reduction in STT of -0.2 mm (min-max: 0.8 to -0.9 mm) at site 1 during the study period, but we did not observe this reduction in all animals. At site 2, we found an average reduction in STT of -0.6 mm (min-max: 0.8 to -1.3 mm) from 10 d before calving to 5 d after calving, corresponding to a decrease of approximately 15% (min-max: 0 to 31%). We suggest that this reduction was caused by a rotation of the distal phalanx toward the posterior part of the sole horn in the weeks around calving. During the same period, heifer BCS decreased between 0.25 and 0.5 increments in the BCS scale, but we found no correlation between BCS and STT. However, the heifers experienced many changes related to management, feeding, and time budget during the study period, and the rotation of the distal phalanx and reduction of the STT might be explained not only by calving but also by other factors that affect heifers and their claws during the transition period.

Differing planes of pre- and postweaning phase nutrition in Holstein heifers: I. Effects on feed intake, growth efficiency, and metabolic and development indicators.

The objectives of this study were to determine the effects of pre- and postweaning planes of nutrition on feed and metabolizable energy (ME) intake, growth, concentrations of glucose, insulin, insulin-like growth factor 1 (IGF-1), and ß-hydroxybutyrate (BHB) in blood and rumen volatile fatty acids (VFA) from birth to 25 wk of age in Holstein heifers. Heifer calves (n = 36) were randomly assigned to receive either a low (5 L/d of whole milk) or high (10 L/d of whole milk) preweaning plane of nutrition from 1 to 7 wk of age, and either a low (70% concentrate dry total mixed ration) or high (85% concentrate dry total mixed ration) postweaning plane of nutrition from 11 to 25 wk of age. From birth to 25 wk of age, feed intake was recorded daily, and body measures were obtained weekly. Circulating hormone and metabolite concentrations were measured biweekly and total ruminal VFA, fecal starch, and body condition were assessed monthly. Overall, average daily gain and body weight were greater for heifers offered increased planes of nutrition during both the pre- and postweaning phases. Heifers offered the high preweaning plane had greater milk intake (7.7 ± 0.1 vs. 4.5 ± 0.1 L/d) but lower starter intake (0.3 ± 0.04 vs. 0.7 ± 0.04 kg/d) during the preweaning phase than those offered the low plane. High preweaning plane heifers also had greater ME intake from wk 1 to 7, but less ME intake at wk 9 (5.3 ± 0.3 vs. 6.6 ± 0.2 Mcal/kg) than those offered the low plane. Furthermore, overall glucose (118.8 ± 2.9 vs. 110.1 ± 2.9 mg/dL) and IGF-1 (101.6 ± 3.6 vs. 75.9 ± 3.6 ng/mL) concentrations were greater for high versus low preweaning plane heifers, although circulating insulin and BHB did not differ between preweaning plane groups. However, heifers offered the high preweaning plane had reduced total rumen VFA concentrations compared with heifers offered the low plane in the preweaning phase (47.3 ± 2.0 vs. 55.6 ± 2.1 mM). During the postweaning phase, dry matter intake and ME were consistently greater in heifers offered the high postweaning plane. Overall insulin (2.0 ± 0.1 vs. 1.8 ± 0.1 ng/mL), glucose (97.1 ± 0.6 vs. 92.1 ± 0.6 ng/mL), IGF-1 (178.0 ± 4.8 vs. 155.8 ± 4.8 ng/mL), and BHB concentrations (8.8 ± 0.2 vs. 8.1 ± 0.2 ng/mL) were greater in heifers offered the high than the low postweaning plane. In addition, heifers offered the high postweaning plane had increased VFA concentrations in the postweaning phase (73.4 ± 1.3 vs. 63.9 ± 1.3 mM) compared with heifers offered the low postweaning plane. The results indicated that increasing the pre- and postweaning planes of nutrition along with energy levels positively influenced several indicators associated with heifer development before 25 wk of age. Nevertheless, there was limited interaction in growth and development indicators between the 2 phases.

Effect of reproductive management programs for first service on replacement dairy heifer economics.

Our objective was to evaluate cash flow for dairy heifers managed for first service with programs that relied primarily on insemination at detected estrus (AIE), timed AI (TAI), or a combination of both. Holstein heifers from 2 commercial farms were randomized to receive first service with sexed semen after the beginning of the AI period (AIP) at 12 mo of age with 1 of 3 treatments: (1) PGF+AIE (n = 317): AIE after PGF2α injections every 14 d (up to 3) starting at the beginning of the AIP; heifers not AIE 9 d after the third PGF2α were enrolled in the 5d-Cosynch (5dCP) protocol; (2) ALL-TAI (n = 315): TAI after ovulation synchronization with the 5dCP protocol; and (3) PGF+TAI (n = 334): AIE after 2 PGF2α injections 14 d apart (second PGF2α at beginning of AIP). If not AIE 9 d after the second PGF2α, the 5dCP protocol was used for TAI. After first service heifers were AIE or received TAI after the 5dCP with conventional semen. Individual heifer cash flow (CF) for up to a 15-mo period (d 0 = beginning of AIP) was calculated using reproductive cost (rearing only), feed cost (rearing only), income over feed cost (lactation only), calf value, operating cost, and with or without replacement cost. A stochastic analysis with Monte Carlo simulation was used to estimate differences in CF for a range of market values for inputs and outputs. Time to pregnancy for up to 100 d after the beginning of the AIP was analyzed by Cox's proportional regression, binary data with logistic regression, and continuous outcomes by ANOVA. Time to pregnancy (hazard ratio and 95% CI) was reduced for the ALL-TAI versus the PGF+AIE treatment (1.20; 1.02-1.42), but it was similar for ALL-TAI and PGF+TAI (1.13; 0.95-1.33) and the PGF+AIE and PGF+TAI treatments (1.07; 0.91-1.25). The proportion of heifers not pregnant by 100 d did not differ (PGF+AIE = 7.0%; PGF+TAI = 6.5%; ALL-TAI = 6.8%). When including replacement cost, CF ($/slot per 15 mo) differences were $51 and $42 in favor of the PGF+TAI and ALL-TAI compared with the PGF+AIE treatment, and $9 in favor of the PGF+TAI compared with the ALL-TAI treatment but did not differ statistically. Excluding heifers that were replaced to evaluate the effect of timing of pregnancy differences only, the difference in CF between the PGF+AIE with the PGF+TAI and ALL-TAI treatment was the same (i.e., $15) and favored the programs that used more TAI, but also did not differ statistically. Stochastic simulation results were in line with those of the deterministic analysis confirming the benefit of the programs that used more TAI. We concluded that submission of heifers for first service with TAI only or TAI in combination with AIE generated numerical differences in CF of potential value to commercial dairy farms. Reduced rearing cost and increased revenue during lactation increased CF under fixed (not statistically significant) or simulated variable market conditions.