Preweaned heifer management on US dairy operations: Part III. Factors associated with Cryptosporidium and Giardia in preweaned dairy heifer calves

The objective of this study was to evaluate management practices and environmental factors associated with cryptosporidiosis and giardiasis in preweaned heifer calves on US dairy operations. This study was conducted as part of the calf component of the National Animal Health Monitoring System's Dairy 2014 study. The calf component included 104 dairy operations in 13 states and was an 18-mo longitudinal study focused on dairy heifer calves from birth to weaning. Fecal samples were collected from 2,249 calves: 839 calves in the West region (California, Colorado, and Washington) and 1,410 calves in the East region (Iowa, Michigan, Minnesota, Missouri, New York, Ohio, Pennsylvania, Vermont, Virginia, and Wisconsin). Fecal samples were collected only once from calves during the preweaning period. Samples were collected from calves 3 to 66 d of age, with a mean of 22 d. Overall, Cryptosporidium and Giardia were detected in 43.1 and 30.5% of fecal samples, respectively. Backward elimination logistic model selection was used after univariate screening to determine which management practices and environmental factors significantly affected the presence of Cryptosporidium or Giardia. The final Cryptosporidium model included herd size, days of age at fecal collection, and average temperature-humidity index for the month of fecal collection (fTHI). Cryptosporidium was found on a higher percentage of large operations (≥500 cows) than small operations (30 to 99 cows). Younger calves were more likely to have a fecal sample positive for Cryptosporidium than samples from older calves. Fecal samples from calves during the warmer parts of the year (fTHI >70) were more likely to be positive for Cryptosporidium than samples collected in colder months (fTHI <20). The final Giardia model included herd size, days of age at fecal collection, average fTHI, failure of passive transfer status, and average daily gain (kg/d) during the preweaning period. Giardia was isolated more frequently from calves on small operations than on large operations and from calves that were older compared with younger calves. Giardia was more frequently isolated in warmer months. Samples from calves with failure of passive transfer were more likely to have Giardia than calves with adequate passive transfer (>10 g/L IgG). Average daily gain during the preweaning period was lower in calves from which Giardia was isolated. These results highlight the factors associated with the presence of Cryptosporidium and Giardia in preweaned dairy heifer calves.     

Evaluation of factors associated with immunoglobulin G,fat, protein,and lactose concentrations in bovine colostrum and colostrum management practices in grassland-based dairy systems in Northern Ireland

The objectives of this study were to investigate colostrum feeding practices and colostrum quality on commercial grassland-based dairy farms, and to identify factors associated with colostrum quality that could help inform the development of colostrum management protocols. Over 1 yr, background information associated with dairy calvings and colostrum management practices were recorded on 21 commercial dairy farms. Colostrum samples (n = 1,239) were analyzed for fat, protein, lactose, and IgG concentration. A subset was analyzed for somatic cell count and total viable bacteria count. Factors associated with nutritional and IgG concentrations were determined using both univariate and multivariate models. This study found that 51% of calves were administered their first feed of colostrum via esophageal tube, and the majority of calves (80%) were fed >2 L of colostrum at their first feed (mean = 2.9 L, SD = 0.79), at a mean time of 3.2 h (SD 4.36) after birth, but this ranged across farms. The mean colostral fat, protein, and lactose percentages and IgG concentrations were 6.4%, 14%, 2.7%, and 55 mg/mL, respectively. The mean somatic cell count and total viable count were 6.3 log₁₀ and 6.1 log₁₀, respectively. Overall, 44% of colostrum samples contained <50 mg/mL IgG, and almost 81% were in excess of industry guidelines (<100,000 cfu/mL) for bacterial contamination. In the multivariate model, IgG concentration was associated with parity and time from parturition to colostrum collection. The nutritional properties of colostrum were associated with parity, prepartum vaccination, season of calving, and dry cow nutrition. The large variation in colostrum quality found in the current study highlights the importance of routine colostrum testing, and now that factors associated with lower-quality colostrum on grassland-based dairy farms have been identified, producers and advisers are better informed and able to develop risk-based colostrum management protocols.     

Short communication: Effect of barn climate and management-related factors on bovine colostrum quality

Several factors have been reported to influence colostrum quality (immunoglobulin concentration). To date, knowledge of the influence of climatic factors in association with other potential influencing factors on colostrum quality is scarce. Associated influential factors are parity, body condition score, length of dry period, ration fed ante partum (AP), β-hydroxybutyrate postpartum (PP), milk yield, milk fat and protein, as well as somatic cell counts from previous and current lactation. The objective of the present study was to examine the effect of barn climate and the aforementioned factors on colostrum quality. Data were collected from 1,381 multiparous Holstein Friesian cows kept on one dairy farm over a period of one year (August 2014 to August 2015). Colostrum was harvested on farm within 1 h PP. The quantity and quality of first colostrum (estimated by Brix refractometry) were recorded for each cow. Additional data recorded were parity, body condition score at drying off, length of dry period, ration fed AP, milk yield data from previous and current lactation, milk somatic cell counts, and β-hydroxybutyrate PP. During the study period, temperature and humidity were recorded in the barn every hour, and temperature-humidity-index (THI) was calculated. Linear regression was performed with colostrum quality as the dependent variable. In the final model, colostrum quantity (L), length of dry period, parity, and climatic factors (specifically, median humidity in the 3rd week AP and hours with THI ≥72 in the last 14 and 21 d AP, respectively) were significant. Colostrum quality improved with parity and length of dry period and decreased with colostrum quantity, humidity, and hours with a THI ≥72. A classification and regression tree analysis revealed that colostrum quantity was the most important factor in this model [normalized importance (NI) 100%]. Parity (NI 42.7%), length of dry period (NI 37.1%), and climatic factors (NI 0.4 to 1.9%) followed with decreasing importance. These results indicate that the most important factors for colostrum quality (i.e., colostrum quantity and parity) may not be influenced by management. The 2 factors that can be influenced by management [i.e., length of dry period and THI (e.g., by cooling)], were quantitatively of minor importance compared with the other 2 factors. Further studies are necessary to determine whether changing these factors can improve colostrum quality significantly.     

Short communication: Processed bovine colostrum milk protein concentrate increases epithelial barrier integrity of Caco-2 cell layers

Colostrum plays an important role in initiating the development of the intestinal barrier in newborn mammals. Given its bioactivity, there is much interest in the potential use of bovine colostrum to improve human gastrointestinal health throughout the life span. There is evidence that bovine colostrum is effective at improving small intestinal barrier integrity and some indication that it may alter colonic motility. However, for colostrum to be used as a product to improve intestinal health, it needs to be bioactive after processing. The aim of this study was to determine whether industrial processing of bovine colostrum affects its ability to improve small intestinal barrier integrity or alter distal colon motility. Three colostrum sample types were compared; raw whole colostrum powder (WCP), raw skim colostrum powder (SCP), and industrially produced colostrum milk protein concentrate (CMPC). To determine whether these colostrum powders had different effects on small intestinal barrier integrity, their effects on the transepithelial electrical resistance across an in vitro intestinal epithelial layer (Caco-2 cells) were measured, both with and without a challenge from the proinflammatory cytokine tumor necrosis factor-α. These results showed that CMPC enhanced transepithelial electrical resistance across unchallenged epithelial cell layers, whereas the raw colostrum samples, WCP and SCP, did not have an effect. The colostrum samples were also compared to determine how they affect contractility in the distal colon isolated from the rat. Skim colostrum powder was the only sample to act directly on colonic tissue to modulate motility, increasing the amplitude of contractions. The results show that bovine colostrum is able to improve small intestinal barrier integrity and alter colon motility, and they implicate different components. The barrier integrity enhancement was apparent only in the industrial CMPC, which may have been due to the increase in protein concentration or the release of small peptides as a result of processing. The ability to alter colon motility was present in SCP but absent in WCP, again implying that an increase in protein concentration is responsible for the effect. However, this effect was not apparent for the industrially processed CMPC, suggesting denaturation or degradation of the active component. The beneficial effect of colostrum on small intestinal barrier integrity was present after processing, confirming that it is feasible to industrially produce an active product for gut health.     

Short communication: Validation of a novel milk progesterone-based tool to monitor luteolysis in dairy cows using cost-effective,on-farm measured data

The progesterone (P4) monitoring algorithm using synergistic control (PMASC) uses luteal dynamics to identify fertility events in dairy cows. This algorithm employs a combination of mathematical functions describing the increasing and decreasing P4 concentrations during the development and regression of the corpus luteum and a statistical control chart that allows identification of luteolysis. The mathematical model combines sigmoidal functions from which the cycle characteristics can be calculated. Both the moment at which luteolysis is detected and confirmed by PMASC, as well as the model features themselves, can be used to inform the farmer on the fertility status of the cows.     

Behavioral and patho-physiological response as possible signs of pain in dairy cows during Escherichia coli mastitis: A pilot study

Bovine mastitis is one of the most common diseases in the dairy industry and it is a major welfare problem. Pain during mastitis is generally assessed through behavior but a combination of indicators would increase the chances of detecting pain and assessing its intensity. The aim of this study was to assess behavioral and patho-physiological responses as possible signs of pain experienced by cows after experimental intramammary challenge (mastitis) with Escherichia coli. Six Holstein-Friesian cows received an inoculation of E. coli P4 in one healthy quarter. Evolution of the disease was assessed using bacteriological growth and somatic cell counts (SCC). Cows' response to the challenge was monitored by direct behavioral and clinical observations, data loggers, rumen temperature sensors, and indicators of inflammation, stress, and oxidative status. From all data recorded, the variables that contributed most to the discrimination of mastitis phases were obtained by factorial discriminant analysis. Baseline levels of all indicators corresponded to values before challenge. Specifically, we weighted data relating to lying behavior by the observations at the same hour of the day before challenge to eliminate the circadian rhythm effect. We identified 3 phases that were discriminated by factorial discriminant analysis with good performance. Nine indicators varied according to the phase of the disease: cows' attitude toward their surroundings, tail position, clinical signs, ear position, variation of postural changes, concentrations of haptoglobin and serum amyloid A (SAA), cortisol blood levels, and rumen temperature (as a surrogate for body temperature). In phase 1 (4 to 8 h postinoculation), E. coli proliferated exponentially in milk but inflammation indicators remained at baseline levels. Cows were less attentive toward their surroundings (median score, 0.63), and postural changes (lying/standing) were less frequent (0.75 times from baseline). In phase 2 (12 to 24 h postinoculation), bacterial concentrations peaked around 12 h and then began to decrease concomitantly with a sharp SCC increase. Cows were less attentive toward their surroundings (score, 0.54), had high plasma cortisol (31.3 ng/mL) and SAA (100.3 µg/mL) concentrations, and rumen temperature was increased (40.3°C). In phase 3 (32 to 80 h postinoculation), bacterial concentrations decreased concomitantly with high SCC levels. Cows had high levels of haptoglobin (0.57 mg/mL) and SAA (269 µg/mL) but showed no behavioral changes. Dairy cows displayed changes of behavioral, inflammatory, and stress parameters after E. coli mammary inoculation. Our results suggest that cows may have experienced discomfort in the preclinical phase (phase 1) and pain in the acute phase (phase 2) but neither discomfort nor pain in the remission phase (phase 3). Although larger controlled studies are needed to confirm our findings, this knowledge could be useful for early detection of E. coli mastitis and for decision-making regarding the initiation of pain-relief treatment during mastitis in dairy cows. This would improve animal welfare and potentially faster disease remission.     

Short communication: Use of a digital refractometer in assessing immunoglobulin G concentrations in colostrum and the first 5 transition milkings in an Irish dairy herd

Transition milk is a source of immunoglobulin G (IgG) and could potentially be used to provide calves with passive immunity, when the IgG concentration is ≥50 g/L. Assessment of IgG concentrations in transition milk would be required before feeding and could be conducted using cow-side tests such as refractometers. Currently, limited information is available on the ability of refractometers to assess transition milk quality. We hypothesized that digital refractometry could be used to provide an accurate cow-side assessment of IgG concentrations in colostrum and transition milk, and IgG concentration in colostrum and one or more transition milking in an Irish herd is >50 g/L. The objectives of this study were to determine the IgG concentrations in colostrum and first, second, third, fourth, and fifth transition milk, and determine the utility of a digital refractometer in assessing quality of colostrum and transition milk produced by cows in a pasture-based dairy production system. A convenient sample of 75 dairy cows were enrolled. Colostrum and transition milk IgG concentrations were determined by radial immunodiffusion and refractometry. Sensitivity and specificity of the refractometer were determined and cut-off points that maximized sensitivity and specificity were determined using receiver operating characteristic curves. Median (range) IgG concentrations in colostrum and first, second, third, fourth, and fifth milking were 99.6, 43.5, 12.5, 5.3, 1.9, and 1.8 g/L, respectively. The sensitivity (0.8–1) of digital refractometry in identifying samples with low IgG concentrations in colostrum, first, second, and third transition milk was acceptable. In contrast, digital refractometry was not useful for assessing IgG concentrations in the fourth and fifth milking due to low IgG concentrations.     

Short communication: Molecular characterization and antimicrobial resistance of pathogenic Escherichia coli isolated from raw milk and Minas Frescal cheeses in Brazil

The aim of this study was to quantify, identify, evaluate antimicrobial resistance, and characterize the virulence factors of enteropathogenic (EPEC), Shiga-toxigenic (STEC), and enterohemorrhagic (EHEC) Escherichia coli in raw milk (RM) and legal (LMFC) and illegal (IMFC) Minas Frescal cheeses in southern and northeast Brazil. Illegal cheeses are those made without official inspection service or sanitary surveillance. We evaluated samples of RM produced in Paraná (southern) and Maranhão (northeast) States, LMFC produced using pasteurized milk in inspected industries, and IMFC potentially produced with raw milk. Mean total coliform counts were 8.4 × 10⁴ cfu/mL for RM, 1.4 × 10⁷ cfu/mL for LMFC, and 2.9 × 10⁷ cfu/mL for IMFC. Mean E. coli counts were 2.4 × 10³ cfu/mL for RM, 1.9 × 10² cfu/mL for LMFC, and 1.1 × 10⁵ cfu/mL for IMFC. Among the 205 E. coli isolates from RM, 9.75% were identified as EPEC, mainly (90%) in samples from Paraná. Of the total isolates from the cheese samples, 97.4% (n = 111) came from IMFC, of which 1.8 and 2.7% were identified as EPEC and STEC, respectively; no EHEC was detected. The phylogenetic group A (60%) and typical EPEC (68%) predominated, which confirms the possible human origin of pathogenic isolates in RM and IMFC. Of these, 50% were resistant to at least one antibiotic, and streptomycin was the antimicrobial with the highest number (8) of EPEC and STEC resistant isolates. This study reports the first isolation of serogroup O28ac in Brazilian milk. We found no predominance of a specific serogroup of EPEC or STEC in milk or cheese or clonal isolates in the same sample, indicating different origins of the contamination in these products, presumably mostly related to poor hygienic handling.     

Short communication: Reproduction outcomes in dairy heifers following a 14-d progesterone insert presynchronization protocol

The objectives were to evaluate pregnancy per artificial insemination (AI), days to first AI, and proportion pregnant within 7 d of AI eligibility in dairy heifers subjected to presynchronization compared with dairy heifers not presynchronized. Thirty days before AI eligibility, Holstein heifers were assigned randomly to 1 of 3 groups: 14-d controlled internal drug release (CIDR; containing progesterone) presynchronization, PGF_2α presynchronization, or control (no presynchronization). Heifers in the 14-d CIDR presynchronization treatment (n = 119) received a CIDR on d ?30, which was removed on d ?16, followed by an injection of PGF_2α upon entry to the breeding program (d 0). Heifers in the PGF_2α presynchronization treatment (n = 118) received an injection of PGF_2α on d ?11 and d 0. Control heifers (n = 121) were not presynchronized and received an injection of PGF_2α on d 0. All heifers received tail paint on d 0 to facilitate once-daily detection of estrus (based on paint removal). Heifers detected in estrus received AI with conventional semen on the same morning as detected estrus. Generalized linear mixed models were used to assess mean treatment differences. Following PGF_2α treatment on d 0, more heifers were detected in estrus in the first 7 d after eligibility in the 14-d CIDR group (95.8%) compared with the PGF_2α (74.6%) and control (66.9%) groups. Days to first AI differed between treatments (14-d CIDR = 3.6 d vs. PGF_2α = 5.0 d vs. control = 6.8 d). Pregnancy per AI for first AI within 7 d of eligibility was 71.9% (14-d CIDR), 58.0% (PGF_2α), and 61.7% (control), and differed between 14-d CIDR and PGF_2α heifers. Presynchronization with a 14-d CIDR increased the proportion of heifers pregnant in the first 7 d of eligibility (14-d CIDR = 68.9% vs. PGF_2α = 43.2% vs. control = 41.3%). Projected days on feed (d 0 to projected calving date) were 295 (14-d CIDR), 302 (PGF_2α), and 305 (control), and were different between the 14-d CIDR and control heifers. The potential economic benefit to the producer was $15.85 per heifer presynchronized with a 14-d CIDR protocol compared with the control group. Treatment of dairy heifers with a 14-d CIDR effectively presynchronized estrus, resulting in a greater proportion detected in estrus, reduced days to first AI, and an increased proportion of heifers pregnant within the first 7 d after breeding eligibility compared with heifers presynchronized with a single PGF_2α injection and control heifers.     

Short communication: Supplementation of colostrum and milk with 5-hydroxy-l-tryptophan affects immune factors but not growth performance in newborn calves

In ruminants, colostrum is the main source of immunoglobulins for the newborn animal, conferring immune protection until the immune system becomes active and able to synthesize its own immunoglobulins. Serotonin (5-HT), a biogenic amine derived from tryptophan, has stimulatory effects on many physiological processes, including components of the innate (mastocytes, eosinophils, and natural killer cells) and adaptive (T and B lymphocytes) immune systems. Based on the known effects of 5-HT on the immune system, we hypothesized that increased concentrations of 5-HT, through administration of its precursor 5-hydroxy-l-tryptophan (5-HTP), may positively affect development of the calf's immune system and therefore support health and growth performance during the first weeks of life. Eighteen calves were randomly assigned to 1 of 2 experimental groups (control and 5-HTP), resulting in n = 9 per treatment group. Both groups received 2 colostrum meals from a common pool of colostrum. Thereafter, calves were fed milk replacer twice daily for 30 d. In the 5-HTP group, colostrum and milk replacer were supplemented with 1.5 mg of 5-HTP/kg of birth weight during the first 15 d after birth. Body weight was recorded at birth and on d 5, 10, 15, and 30 after birth. Blood samples were collected every morning (0800 h) before feeding from birth until d 5 and then on d 7, 9, 11, 13, 15, and 30 after birth. Serum 5-HT concentrations were increased as a consequence of the 5-HTP supplementation. Plasma immunoglobulin G concentrations did not differ between groups throughout the experimental period. The blood mRNA abundance of several factors related to the innate and adaptive immune system [nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), serum amyloid A-1 (SAA1), chemokine C-C motif ligand 5 (CCL5), cyclooxygenase 2 (PTGS2), haptoglobin (HP), and IL-1β] were increased in calves supplemented with 5-HTP. Supplementation of 5-HTP did not affect any of the measured metabolites (fatty acids and glucose) or minerals (calcium and magnesium) or milk feed intake, feed conversion ratio, and growth. In conclusion, 5-HTP supplementation induced an increase of 5-HT concentrations in blood and caused an increase in mRNA abundance of several factors related to the innate and adaptive immune systems, which might increase the protection of the calf against external agents.     

Short communication: Assessing the accuracy of inline milk fat-to-protein ratio data as an indicator of hyperketonemia in dairy cows in herds with automated milking systems

The objective of this study was to evaluate the accuracy of inline milk fat-to-protein (F:P) data to detect hyperketonemia (HYK) in herds with automated milking systems (AMS). The F:P ratio has been investigated as a tool for detecting HYK with moderate accuracy in past studies, but inline F:P data in AMS may also be useful for HYK screening. To assess the accuracy of these data in commercial settings, we monitored 484 cows from 9 AMS herds for their first 3 wk of lactation, taking blood samples once per week (n = 1,427). Positive cases of HYK were defined by whole-blood β-hydroxybutyrate (BHB) concentrations ≥1.2 or ≥1.4 mmol/L. Milk data were collected from the AMS software on each farm for each cow and converted into 4 different F:P values: (1) value from the same day as the BHB test; (2) 5-d centered-moving average (CMA); (3) 5-d backward-moving average (BMA); (4) 5-d forward-moving average (FMA). In linear regression models, all 4 values were associated with BHB, but slope estimates varied and R² were low: same day (slope = 0.95, R² = 0.07), CMA (slope = 1.05, R² = 0.07), BMA (slope = 0.65, R² = 0.04), and FMA (slope = 1.23, R² = 0.09). In logistic regression models, the odds of having HYK (BHB ≥1.2 mmol/L) increased with every 0.1-unit increase from the mean F:P ratio (1.16) using same-day values (odds ratio = 1.35, 95% confidence interval = 1.25–1.47) and CMA (odds ratio = 1.39, 95% confidence interval = 1.27–1.51). The same increase in F:P from mean BMA (1.14) and FMA (1.17) was associated with 1.22 and 1.49 times the odds of HYK, respectively. For all 4 F:P variations, we evaluated the sensitivity, specificity, positive predictive value, and negative predictive value of different F:P thresholds with HYK status. As the F:P threshold increased from 1.17 to 1.50, sensitivity decreased (range: 77 to 9%) but specificity increased (range: 58 to 96%). Same-day and CMA F:P cutoffs at which a balance was reached between sensitivity and specificity ranged from 1.18 to 1.22; however, even at these values we found high rates of false positives and negatives (range: 31–39%). These results suggest that inline milk F:P data from inconsistently calibrated sensors should not be used alone to detect HYK in AMS herds.     

Short communication: Diagnostic performance of on-farm bacteriological culture systems for identification of uterine Escherichia coli in postpartum dairy cows

The objectives of this study were to validate the performance of on-farm bacteriological culture systems for identification of Escherichia coli in the uterus of early postpartum dairy cows and to determine if an association is present between the results and the subsequent occurrence of puerperal metritis (PM). A prospective cohort study was conducted in one commercial Holstein dairy herd in which 400 cows were sampled between 24 and 48 h after parturition. Three bacteriological samples were obtained from the uterus of each cow and were cultured for identification of E. coli. One sample was cultured in a commercial bacteriology laboratory according to standard procedures for identification of E. coli and was considered as the reference test. The two other samples were cultured on the farm using the Tri-Plate (University of Minnesota, St. Paul, MN) and Petrifilm systems, and plate readings were done after 24 and 48 h of incubation (variables: Tri24h, Tri48h, Petri24h, Petri48h). Participating cows were followed until 21 days in milk to diagnose PM. The prevalence of PM and E. coli (from the reference test) in the cow population was 15.0 and 33.5%, respectively. Both on-farm culture systems were accurate compared with the reference test. The sensitivity and specificity were 97 and 100%, 99 and 100%, 100 and 92%, and 100 and 89% for Tri24h, Tri48h, Petri24h, and Petri48h, respectively. On-farm results for Tri24h, Tri48h, Petri24h, and Petri48h were associated with subsequent occurrence of PM. The results from this study support the use of the Tri-Plate and Petrifilm culture systems on dairy farms to identify the presence of E. coli in the uterus of postpartum cows.     

Short communication: Absorption of 2-hydroxy-4-methylthiobutanoate in dairy cows

The objective of this study was to measure net portal absorption of 2-hydroxy-4-methylthiobutanoate (HMTBA) in dairy cows. Four multicatheterized lactating cows were used in a cross-over design with 7-d experimental periods. They were fed every other hour a total mixed ration and received, in addition, twice a day, 12.5 or 25 g/meal of HMTBA. On the last day of treatment, net portal absorption of HMTBA was numerically greater after the 25-g compared with the 12.5-g bolus meal of HMTBA, and the amount absorbed relative to the dose ingested was unchanged between treatments averaging 11.2 ± 4.7% of the dose. This represents a minimum value of HMTBA availability because it does not take into account any HMTBA metabolized to Met by gut tissues (in sheep this amounted to another 5% of the dose). A rapid method to estimate net portal absorption based on temporal variations of the peripheral plasma HMTBA concentrations following the HMTBA meal is also presented. Based on the good relationship (concordance correlation coefficient = 0.97) observed between direct measurements and estimations, this simplified approach offers a reasonable approach to assess HMTBA absorption under different feeding situations.     

Short communication: Ketosis,feed restriction,and an endotoxin challenge do not affect circulating serotonin in lactating dairy cows

Circulating serotonin (5-hydroxytryptamine; 5-HT) appears to be associated with various energetic disorders and hypocalcemia during the transition period. The objective of this study was to evaluate the effects of ketosis, feed restriction (FR), and endotoxin challenge (models in which energetic and calcium metabolism are markedly altered) on circulating 5-HT in lactating Holstein cows. Blood samples were obtained from 3 separate experiments; circulating β-hydroxybutyrate (BHB), nonesterified fatty acids (NEFA), and glucose were measured in all 3 experiments, whereas ionized calcium (iCa²⁺) was measured only in the endotoxin challenge. In the ketosis study, blood samples from cows clinically diagnosed with ketosis (n = 9) or classified as healthy (n = 9) were obtained from a commercial dairy farm at d ?7, 3, and 7 relative to calving. Ketosis was diagnosed using a urine-based test starting at 5 d in milk. There was no effect of health status on circulating 5-HT and no association between 5-HT and BHB, NEFA, or glucose; however, 5-HT concentrations progressively decreased following calving. In the FR experiment, mid-lactation cows were either fed ad libitum (n = 3) or restricted to 20% of their ad libitum intake (n = 5) for 5 d. There were no FR effects on circulating 5-HT, nor was FR correlated with energetic metabolites. In the immune activation model, mid-lactation cows were intravenously challenged with either lipopolysaccharide (LPS; 1.5 µg/kg of BW; n = 6) or sterile saline (control; n = 6). Administering LPS decreased (56%) blood iCa²⁺ but had no effect on circulating 5-HT, nor was there a correlation between circulating 5-HT and NEFA, BHB, or iCa²⁺. Circulating 5-HT tended to be positively correlated (r = 0.54) with glucose in Holstein cows administered LPS. In summary, in contrast to expectations, circulating 5-HT was unaffected in models of severely disturbed energetic and Ca²⁺ homeostasis.     

Short communication: Relationship of blood DNA methylation rate and milk performance in dairy cows

The objective of this study was to investigate the global methylation rate in blood DNA and its relationship with lactation performance. A total of 196 mid-lactation dairy cows were fed the same diet under the same management. Milk yield was recorded and blood samples were collected from the jugular vein before morning feeding. The blood global DNA methylation rates were quantified using a methylation quantification kit. Overall, the average blood global DNA methylation rate of all cows was 12.4%. When DNA methylation rates were compared between cows with high (n = 40; 37.0 to 42.0 kg/d) and low (n = 33; 24.0 to 30.0 kg/d) milk yield, DNA methylation rates in the lower-yield cows (14.1 ± 0.7%) were significantly higher than those in the higher-yield animals (11.6 ± 0.7%). Our results indicated an association of milk and protein yields with global DNA methylation rates in lactating dairy cows. However, further research is needed to determine whether this association reflects the true influence of epigenetic mechanisms on yield or whether other factors, such as different proportions of blood cell types in high- and low-yielding cows, affect apparent global DNA methylation levels.     

Effects of postpartum milking strategy on plasma mineral concentrations and colostrum,transition milk,and milk yield and composition in multiparous dairy cows

The effects of postpartum milking strategy on plasma mineral concentrations, blood β-hydroxybutyrate (BHB) concentration, and colostrum, transition milk, and first monthly test milk yield and composition were evaluated in 90 multiparous Jersey and Jersey × Holstein crossbreed cows from a commercial farm. Before first postpartum milking, cows were randomly assigned to the following milking strategies, implemented during the first 2 d postpartum: twice-a-day milking (M2, standard industry practice, milking every 12 h; n = 22), once-a-day milking (M1, milking every 24 h; n = 24), restricted milking (MR, 3-L milking every 12 h; n = 21), and delayed milking (MD, no milking for the first 24 h, and milking every 12 h afterward; n = 23). Blood samples for total plasma Ca, P, and Mg determination were collected from enrollment every 4 h up to 48 h, and at 3 d in milk. Blood BHB concentration was determined at 3 and 11 d in milk. Colostrum and transition milk yields were recorded, and samples were collected at each study milking for IgG and somatic cell count (SCC) determinations. Information for first monthly test milk yield and composition was obtained from the Dairy Herd Improvement Association. Statistical analyses were conducted using generalized multiple linear and Poisson regressions with Dunnett adjustment and M2 as reference group for mean comparisons. Overall, plasma Ca concentration within 48 h after enrollment was higher for MD (2.17 mmol/L), tended to be higher for MR (2.15 mmol/L), and was similar for M1 (2.09 mmol/L) compared with M2 cows (2.06 mmol/L). No statistically significant differences compared with M2 cows were observed for plasma P and Mg concentrations. Colostrum and transition milk and total Ca harvested within 48 h after enrollment were lower for M1, MR, and MD compared with M2 cows. The MD strategy prevented harvesting colostrum with >50 g of IgG/L. No statistically significant effects were detected on plasma mineral concentrations at 3 DIM, blood BHB concentration, colostrum and transition milk SCC within 48 h after enrollment, or milk yield, energy-corrected milk yield, and SCC at first monthly test. Our results suggest that postpartum plasma Ca concentration may be influenced by postpartum milking strategy, without interfering with future milk yield and udder health. Further studies should evaluate whether the proposed milking strategies in early postpartum affect production, reproduction, or health.     

Non-aureus staphylococci in fecal samples of dairy cows: First report and phenotypic and genotypic characterization

The aims of this study were to determine whether non-aureus staphylococci (NAS) are present in rectal feces of healthy dairy cows, and if so, to delineate species to which they belong and to study several phenotypic and genotypic traits as a first step toward determining the potential impact of fecal shedding of NAS on bovine udder health. Fecal samples were aseptically collected from the rectum of 25 randomly selected clinically healthy dairy cows in a commercial dairy herd using an automated milking system. Fecal NAS were isolated and then identified at the species level using transfer RNA-intergenic spacer PCR and sequencing of the 16S rRNA housekeeping gene. Strain typing was performed using random amplification of polymorphic DNA (RAPD)-PCR. The antimicrobial resistance profiles, biofilm formation, and growth and inhibitory characteristics of all NAS isolates were evaluated. Half of the cows were shedding NAS, resulting in 31 NAS isolates belonging to 11 different species. The most prevalent species were Staphylococcus rostri (23%, n = 7), Staphylococcus cohnii (16%, n = 5), and Staphylococcus haemolyticus (13%, n = 4) with all Staphylococcus agnetis, Staphylococcus chromogenes, and Staph. rostri isolates belonging to the same strain according to RAPD banding patterns. Acquired antimicrobial resistance was observed in 28 of the 31 NAS isolates, mainly due to β-lactamase production. Most of the isolates (84%, n = 27) had a weak biofilm-forming potential, but only 2 contained the bap gene. The ica and aap genes were not detected in any of the isolates. In vitro growth of Staphylococcus aureus and Streptococcus dysgalactiae was inhibited by Staph. agnetis isolates, and Staph. chromogenes isolates were able to inhibit the growth of Strep. dysgalactiae and Streptococcus uberis. All fecal isolates were able to grow when oxygen and iron were limitedly available, mimicking the growth conditions in the mammary gland.     

Short communication: Production performance and nutrient digestibility of lactating dairy cows fed diets with and without addition of a live-yeast supplement

The objective of this study was to evaluate the use of a live-yeast product when feeding relatively high-forage diets to high-producing cows in mid lactation. Eight primiparous [607 ± 43 kg of body weight (BW) and 130 ± 16 d in milk (DIM) at the beginning of the experiment] and 16 multiparous (706 ± 63 kg of BW and 137 ± 22 DIM at the beginning of the experiment) Holstein cows were blocked by parity and DIM, and randomly assigned to 1 of 2 diets (control vs. yeast) for a 12-wk period according to a randomized complete block design. The formulated diets contained 50.4% corn silage, 10.4% alfalfa hay, and 39.2% concentrate. The yeast diet was formulated to provide approximately 5.4 × 10¹¹ cfu/d of Saccharomyces cerevisiae (BeneSacc; Global Nutritech Biotechnology LLC, Richmond, VA). Total-tract nutrient digestibility was estimated using 240-h undigested neutral detergent fiber (NDF) as an internal marker. Supplementing live yeast to lactating dairy cows did not affect dry matter intake (25.0 kg/d), milk yield (38.6 kg/d), milk fat concentration (4.78%), milk fat yield (1.83 kg/d), milk protein concentration (3.09%), milk protein yield (1.18 kg/d), milk lactose concentration (4.79%), milk lactose yield (1.84 kg/d), BW gain (?0.05 kg/d), or body condition score gain (0.16 units). The digestibility of dry matter was greater for the control treatment than for the yeast treatment (69.3 and 67.1%, respectively), but the digestibilities of crude protein (61.5%), NDF (40.5%), and starch (98.6%) were not affected by treatment. In conclusion, supplementation of live yeast did not affect production performance or nutrient digestibility of high-producing cows in mid lactation. The reasons for the lack of effect are not clear, but an evaluation of interactions between yeast and rumen buffer supplementation is warranted.     

Short communication: Comparative estimation of colostrum quality by Brix refractometry in bovine,caprine, and ovine colostrum

Newborn ungulates depend on the timely supply of colostrum containing sufficient immunoglobulins to obtain passive immunity against disease. Brix refractometry enables a rapid on-farm estimation of colostrum quality and has been intensively studied in bovines. However, the suitability of Brix refractometers for assessing colostrum quality in goats and ewes has been scarcely evaluated. The present study compared bovine, caprine, and ovine colostrum quality estimation using an optical Brix refractometer. In addition, between-species variations in the relationships between Brix values and colostrum constituents (IgG, fat, protein, and lactose) and the accuracy of Brix refractometry at different cutoff values were evaluated by a receiver operating characteristic curve analysis. We measured the Brix value and contents of IgG, fat, protein, and lactose in 324 colostrum samples (108 cows, 116 does, and 100 ewes). Thresholds for classification of good colostrum quality (as determined by ELISA) were set at 50 mg IgG/mL in cows and 20 mg/mL in does and ewes. Bovine colostrum showed the greatest IgG concentrations compared with caprine and ovine colostrum. Fat and protein content was higher in sheep colostrum compared with the other species, whereas the highest lactose concentrations were detected in goat colostrum. Brix values ranged from 11.4 to 34.6% (22.1 ± 4.2%; mean ± standard deviation), 15.4 to 40.0% (28.5 ± 6.8%), and 8.8 to 39.8% (21.6 ± 5.3%) in bovine, ovine, and caprine colostrum, respectively. In all 3 species, Brix was highly correlated with IgG and protein concentrations (cows, r = 0.83 and 0.98; goats, r = 0.83 and 0.89; sheep, r = 0.75 and 0.87). Optimal cutoff points for greatest accuracy of Brix measurements were 19.3% Brix in cows [with 87.1% sensitivity (Se) and 100% specificity (Sp)], 20.7% Brix in does (with 53.5% Se and 100% Sp), and 26.5% Brix in ewes (with 75% Se and 91.3% Sp). In conclusion, Brix refractometry is an acceptable tool for on-farm estimations of colostrum quality in does and ewes despite distinct between-species variations in colostrum composition.