Effects of feeding pasteurized waste milk to dairy calves on phenotypes and genotypes of antimicrobial resistance in fecal Escherichia coli isolates before and after weaning

The aim of this study was to evaluate the effects of feeding pasteurized waste milk (pWM) to calves on antimicrobial resistance of fecal Escherichia coli at both phenotypic and genotypic levels. Fifty-two Holstein female calves (3 ± 1.3 d of age) were fed 1 of the 2 different types of milk: milk replacer (MR) without antimicrobials or pWM with β-lactam residues until weaning at 49 d of age. Fecal swabs of all calves were obtained on d 0, 35, and 56 of the study and 3 E. coli isolates per sample were studied. Phenotypic resistance was tested by the disk diffusion method against a panel of 12 antimicrobials. A total of 13 resistance genes consisting of β-lactam, sulfonamide, tetracycline, and aminoglycoside families were examined by PCR. Feeding pWM to calves increased the presence of phenotypic resistance to ampicillin, cephalotin, ceftiofur, and florfenicol in fecal E. coli compared with MR-fed calves. However, the presence of resistance to sulfonamides, tetracyclines, and aminoglycosides was common in dairy calves independent of their milk-feeding source, suggesting other factors apart from the feeding source are involved in the emergence of antimicrobial resistance.     

Feeding of waste milk to Holstein calves affects antimicrobial resistance of Escherichia coli and Pasteurella multocida isolated from fecal and nasal swabs

The use of milk containing antimicrobial residues in calf feeding programs has been shown to select for resistant fecal Escherichia coli in dairy calves. However, information is scarce about the effects of feeding calves waste milk (WM) on the prevalence of multidrug-resistant bacteria. The objective of this study was to determine the antimicrobial resistance patterns of fecal E. coli and nasal Pasteurella multocida isolates from calves fed either milk replacer (MR) or WM in 8 commercial dairy farms (4 farms per feeding program). Fecal and nasal swabs were collected from 20 ± 5 dairy calves at 42 ± 3.2 d of age, and from 10 of these at approximately 1 yr of age in each study farm to isolate the targeted bacteria. Furthermore, resistance of E. coli isolates from calf-environment and from 5 calves at birth and their dams was also evaluated in each study farm. Resistances were tested against the following antimicrobial agents: amoxicillin-clavulanic acid, ceftiofur, colistin, doxycycline (DO), enrofloxacin (ENR), erythromycin, florfenicol, imipenem, and streptomycin. A greater number of fecal E. coli resistant to ENR, florfenicol, and streptomycin and more multidrug-resistant E. coli phenotypes were isolated in feces of calves fed WM than in those fed MR. However, the prevalence of fecal-resistant E. coli was also influenced by calf age, as it increased from birth to 6 wk of age for ENR and DO and decreased from 6 wk to 1 yr of age for DO regardless of the feeding program. From nasal samples, an increase in the prevalence of colistin-resistant P. multocida was observed in calves fed WM compared with those fed MR. The resistance patterns of E. coli isolates from calves and their dams tended to differ, whereas similar resistance profiles among E. coli isolates from farm environment and calves were observed. The findings of this study suggest that feeding calves WM fosters the presence of resistant bacteria in the lower gut and respiratory tracts of dairy calves.     

Antimicrobial resistance in non-aureus staphylococci isolated from milk is associated with systemic but not intramammary administration of antimicrobials in dairy cattle

Non-aureus staphylococci (NAS) are the bacteria most frequently isolated from bovine milk. Objectives of this study were to determine herd-level associations between antimicrobial use (AMU) and prevalence of antimicrobial resistance (AMR) and antimicrobial resistance genes in NAS according to antimicrobials and routes of administration. The AMR profile was determined using a micro-broth dilution method against a panel of 23 antimicrobials for 1,702 NAS isolates obtained from 89 herds. A subset of these isolates (n = 405) was submitted to whole-genome sequencing, and the presence of AMR genes was determined using data from 4 databases. Antimicrobial use was determined for all herds using an inventory of empty drug containers and quantified for each antimicrobial as the number of antimicrobial daily doses administered. Generalized linear models were used to estimate antimicrobial and route-specific associations between AMR in NAS and AMU. Prevalence of multidrug resistance in NAS was associated with systemic use of antimicrobials. Estimated relative risk associated with a 1-unit increase in antimicrobial daily doses per cow-year administered systemically was 1.28. No association was present with either intramammary or intrauterine use. Three drug classes, all of high or very high importance for human medicine, were associated with drug-specific AMR when administered systemically: penicillins, third-generation cephalosporins, and macrolides. Prevalence of tet, erm, and bla_ARL genes in NAS was higher in herds that used more tetracyclines, macrolides, and third-generation cephalosporins, respectively. No association between drug-specific AMU and prevalence of blaZ, mphC, and msrA was identified, irrespective of route of administration. The either weak or nonexistent association between AMR and antimicrobials administered intramammarily suggest that a decrease in AMR of NAS following implementation of selective dry cow therapy would be minimal in comparison to reduced use of systemic antimicrobials.     

Antimicrobial resistance and prevalence of virulence factor genes in fecal Escherichia coli of Holstein calves fed milk with and without antimicrobials

Diarrhea in calves has a significant effect on the dairy industry. A common management practice for preventing or decreasing the effects of such disease in preweaned calves is by the use of antimicrobials in milk or milk replacer. In this study, Escherichia coli antimicrobial resistance in fecal samples collected from calves 2 to 8 d of age that had or had not received antimicrobials in the milk and that had or had not signs of diarrhea by inspection of fecal consistency were investigated. Specifically, resistance of E. coli isolates to individual antimicrobials, multiresistance patterns, and presence of virulence factors were analyzed. Escherichia coli isolates were tested for susceptibility to 12 antimicrobials by use of a Kirby-Bauer disk diffusion assay. The study was conducted at 3 farms, 1 administering growth-promoting antimicrobials (GPA) in the milk and 2 not using GPA in the milk (NGPA). All isolates were susceptible to ciprofloxacin and cefepime. From the total isolates tested, 84% (n = 251) were resistant to at least 2 antimicrobials and 81% (n = 251) were resistant to 3 or more antimicrobials. When antimicrobial resistance was compared between GPA and NGPA, it was observed that the GPA group had higher odds of antimicrobial resistance for most of the individual antimicrobials tested. No significant correlation of virulence factors in GPA or NGPA and diarrheic or non-diarrheic (control) fecal samples was found. Of the 32 virulence factors evaluated, 21 were detected in the study population; the incidence of only 1 virulence factor was statistically significant in each of the diarrheic status (diarrheic or non-diarrheic) and treatment status (NGPA or GPA) groups. Phylogenetic analysis based on the nucleotide sequence of the DNA gyrase gene (gyrB) from 31 fecal E. coli isolates revealed 3 main clades.     

Risk assessment modelling of fecal shedding caused by extended-spectrum cephalosporin-resistant Escherichia coli transmitted through waste milk fed to dairy pre-weaned calves

Waste milk feeding is a common practice in dairy operations. Regardless of the benefits of this practice to the dairy farmers, concerns from the potential dissemination of antimicrobial-resistant bacteria through the gut and subsequent shedding by calves into the environment are increasing. In this study, we employed Monte Carlo simulation to assess the risk of shedding extended-spectrum cephalosporin-resistant Escherichia coli (ESC-R E. coli) caused by waste milk feeding in pre-weaned calves using an exponential dose-response model fit to data for E. coli O157:H7 in cattle. Data from pertinent studies were included in our model to predict the risk of shedding. The median (5th and 95th percentiles) for the daily risk of shedding ESC-R E. coli by calves fed only contaminated waste milk was predicted to be 2.9 × 10^?3 (2.1 × 10^?3, 3.7 × 10^?3), representing a median daily risk of 29 out of 10,000 calves shedding ESC-R E. coli due to exclusive feeding of waste milk containing ESC-R E. coli. This median value was reduced by 94% when accounting for the proportion of waste milk that does not contain ESC-R E. coli. The overall risk of shedding ESC-R E. coli through the pre-weaning period for farms that feed waste milk to calves was 5.7 × 10^?3 (2.4 × 10^?3, 1.1 × 10^?2), representing 57 out of 10,000 calves. When accounting for the proportion of farms that do not feed waste milk, the pre-weaning period risk was reduced by 23%. By varying the prevalence of ESC-R E. coli in waste milk using values of 3, 1.5, and 1%, the daily risk of shedding decreased by factors of 50, 65, and 82%, respectively, which supports the reduction of contamination or discontinuation of feeding waste milk containing ESC-R E. coli as major mitigation measures to reduce the risk of shedding caused by ingestion of resistant bacteria. It is anticipated that the effects of antimicrobial residues in waste milk, which was not considered herein due to lack of data, would further increase risks. Although waste milk feeding to calves may be economically beneficial to the dairy farmers, there exists the risk of dissemination of ESC-resistant bacteria into the environment.     

Effect of on-farm use of antimicrobial drugs on resistance in fecal Escherichia coli of preweaned dairy calves

Respiratory disease and diarrhea are the 2 most common diseases that result in the use of antimicrobial drugs in preweaned calves. Because the use of drugs in food animals, including dairy calves, has the potential for generating cross-resistance to drugs used in human medicine, it is vital to propose farm practices that foster the judicious use of antimicrobials while assuring animal health and productivity. The objective of this study was to use dairy farm calf treatment records to identify antimicrobial drug treatments in calves and to evaluate their effects on the prevalence of antimicrobial-resistant Escherichia coli from rectal swabs of preweaned dairy calves. Eight farms from central New York participated in the study, 3 farms using individual pen housing management and 5 farms using group pen housing management. Eligible study farms could not add antimicrobial drugs to the milk fed to preweaned calves and were required to have farm records documenting antimicrobial drug treatment of calves from birth to weaning. Three fecal E. coli isolates per calf were tested for susceptibility to 12 antimicrobial drugs using a Kirby-Bauer disk diffusion assay. A total of 473 calves were sampled, from which 1,423 commensal E. coli isolates were tested. Of the 9 antimicrobial drugs used on study farms, only enrofloxacin was significantly associated with reduced antimicrobial susceptibility of E. coli isolates, although treatment with ceftiofur was associated with reduced susceptibility to ceftriaxone. The median numbers of days from treatment with ceftiofur and enrofloxacin to rectal swab sampling of calves were 16 d (range: 1–39) and 12 d (range: 6–44), respectively. At the isolate level, treatment with enrofloxacin resulted in odds ratios of 2 [95% confidence interval (CI): 1–4] and 3 (95% CI: 2–6), respectively, for isolation of nonsusceptible E. coli to nalidixic acid and ciprofloxacin compared with calves not treated with enrofloxacin. Treatment with ceftiofur resulted in an odds ratio of 3 (95% CI: 0.9–12) for isolation of nonsusceptible E. coli to ceftriaxone compared with calves not treated with ceftiofur. Treatment with enrofloxacin resulted in selection of isolates that presented phenotypic resistance to both ciprofloxacin and ceftriaxone. Treatment with ceftiofur resulted in a higher prevalence of isolates resistant to ≥3 antimicrobial drugs (97%) compared with no treatment with ceftiofur (73%). These findings reinforce the necessity for continued implementation of practices at the dairy farm that support the sustainable and judicious use of antimicrobial drugs in dairy calves.     

Antibiotic resistance in gut bacteria from dairy calves: a dose response to the level of antibiotics fed in milk

Dairy calves are commonly fed milk from cows treated with antibiotics. The concentration of beta-lactam antibiotic residues found in milk from treated cows was used to determine the range of concentrations of penicillin used in a dose-regulated experiment. Thirty-one Holstein calves were randomly assigned to milk with penicillin G added at concentrations of 0, 6.25, 12.5, 25, and 50 microl/kg. Fecal swabs were taken from each calf twice weekly. Resistance to penicillin was tested by measuring the zone of inhibition in bacterial growth around a disk impregnated with the antibiotic. Inhibition was greatest for bacteria from calves fed milk with no penicillin (2.89 +/- 0.14 mm), and declined as the penicillin dose provided in the milk increased (to a low of 0.70 +/- 0.10 for the 50 microl/kg treatment group). In conclusion, resistance of gut bacteria to antibiotics increases with increasing concentrations of penicillin in the milk fed to dairy calves.     

Randomized clinical trial of the effect of a fixed or increasing milk allowance in the first 2 weeks of life on health and performance of dairy calves

The objective of this study was to describe the effect of offering a fixed or increasing milk allowance in the first 1 to 2 wk of life. We hypothesized that calves offered a fixed amount of milk early in life would not experience more scours, but rather would experience improved health and growth compared with calves that had their daily milk allowance slowly increased over a period of 1 to 2 wk. This randomized controlled clinical trial was conducted on 5 dairy farms in Minnesota with both a summer (June–August 2016) and winter (December–February 2017) period of enrollment. Heifer calves were enrolled at birth, weighed, and systematically assigned by birth order to either the slowly increasing (INC) control group or fixed allowance (FIX) treatment group by farm personnel. Calves assigned to the INC group were slowly increased from 4 to 5 L/d to gradually reach the full peak milk allowance of 6 to 8 L/d over a 7- to 14-d period, whereas calves assigned to the FIX group were offered a full peak milk allowance of 6 to 8 L/d beginning on d 1 after birth. The average FIX calf consumed an extra 14 L of milk as compared with INC calves over the first 2 wk of life, corresponding to an average INC intake of 5 L/d during first 1 to 2 wk of life as compared with an average intake of 6.8 L/d in FIX calves. Study technicians visited all farms weekly to collect health and performance data. Multivariable mixed models were used to describe the effect of treatment (INC/FIX) on 3-wk average daily gain (kg/d), 3-wk weight (kg), and hip height at wk 1, 3, and 7, controlling for the effect of season, birth weight, and the random effect of calf within farm. Multivariable logistic regression models were used to describe the effect of treatment on odds of technician and producer reported health events. A total of 1,264 heifer calves were enrolled (FIX n = 641; INC n = 623) with no difference in enrollment weight or hip height between groups. By 3 wk of age, FIX calves weighed 1.4 (0.59) kg more than INC calves, though the magnitude of this difference varied depending on the period of time INC calves were slowly increased in milk allowance (7 vs. 10 vs. 14 d). Calves in the FIX group grew 0.1 kg/d faster and were taller at wk 3 (0.3 ± 0.15 cm) of life. Forty-two percent (536/1,264) of all enrolled calves had a first treatment event, with no effect of treatment on technician-reported health scores and no overall effect on producer-reported treatment or mortality events. Under the conditions of this study, offering a fixed milk allowance from d 1 of life improved calf growth during the first 3 wk as compared with a gradual increase in milk allowance, with no detrimental effect on calf health.     

Feeding behaviors collected from automated milk feeders were associated with disease in group-housed dairy calves in the Upper Midwest United States

Automated milk feeders (AMF) are an attractive option for producers interested in adopting practices that offer greater behavioral freedom for calves and can potentially improve labor management. These feeders give farmers the opportunity to have a more flexible labor schedule and more efficiently feed group-housed calves. However, housing calves in group systems can pose challenges for monitoring calf health on an individual basis, potentially leading to increased morbidity and mortality. Feeding behavior recorded by AMF software could potentially be used as an indicator of disease. Therefore, the objective of this observational study was to investigate the association between feeding behaviors and disease in preweaning group-housed dairy calves fed with AMF. The study was conducted at a dairy farm located in the Upper Midwest United States and included a final data set of 599 Holstein heifer calves. The farm was visited on a weekly basis from May 2018, to May 2019, when calves were visually health scored and AMF data were collected. Calf health scores included calf attitude, ear position, ocular discharge, nasal discharge, hide dirtiness, cough score, and rectal temperatures. Generalized additive mixed models (GAMM) were used to identify associations between feeding behavior and disease. The final quasibinomial GAMM included the fixed (main and interactions) effects of feeding behavior at calf visit-level including milk intake (mL/d), drinking speed (mL/min), visit duration (min), rewarded (with milk being offered) and unrewarded (without milk) visits (number per day), and interval between visits (min), as well as the random effects of calf age in regard to their relationship with calf health status. Total milk intake (mL/d), drinking speed (mL/min), interval between visits (min) to the AMF, calf age (d), and rewarded visits were significantly associated with dairy calf health status. These results indicate that as total milk intake and drinking speed increased, the risk of calves being sick decreased. In contrast, as the interval between visits and age increased, the risk of calves being sick also increased. This study suggests that AMF data may be a useful screening tool for detecting disease in dairy calves. In addition, GAMM were shown to be a simple and flexible approach to modeling calf health status, as they can cope with non-normal data distribution of the response variable, capture nonlinear relationships between explanatory and response variables and accommodate random effects.     

Evaluation of a colostrum supplement, with or without trypsin inhibitor, and an egg protein milk replacer for dairy calves

Forty-eight Holstein bull calves were assigned to a 2 x 2 x 2 factorial arrangement in a completely randomized block design. Main effects were colostrum versus a serum-derived colostrum supplement, 0 versus 1 g of trypsin inhibitor added at the initial 2 feedings, and milk replacer containing 0 or 50% CP from whole egg. Calves were bled at 0, 6, 12, 18, and 24 h after birth for determination of serum immunoglobulin (Ig). G. Serum IgG concentrations were lower in calves consuming the colostrum supplement compared with calves consuming colostrum. Apparent efficiency of absorption of IgG was similar. Trypsin inhibitor did not affect IgG concentrations or absorption of IgG. Calves were fed either milk replacer for 28 to 35 d (preweaning phase) and weaned when they consumed 0.7 kg of starter grain for 2 consecutive days. The postweaning phase was from weaning to d 56. Feeding colostrum supplement resulted in higher fecal scores postweaning (1.90 vs. 1.58) and overall (1.85 vs. 1.65) and fewer days medicated preweaning (5.1 vs. 2.2 d) and postweaning (3.9 vs. 1.9 d) and overall (9.0 vs. 4.2 d). Calves were treated for upper respiratory tract infections and diarrhea. Dry matter intake and weaning age were not affected by treatment. Postweaning (1.69 vs. 1.2 kg) and overall (1.22 vs. 1.0 kg), calves that received colostrum and egg milk replacer consumed more dry matter and starter. Postweaning, calves fed colostrum and egg milk replacer had similar or greater body weight and gains compared with calves fed colostrum and milk protein milk replacer. Preweaning, feed efficiency was greater for calves fed colostrum (0.44 vs. 0.34), trypsin inhibitor (0.42 vs. 0.36), and milk protein milk replacer (0.48 vs. 0.30) compared with calves fed colostrum supplement, no trypsin inhibitor, and egg milk replacer, respectively. Trypsin inhibitor increased feed efficiency postweaning. Calves fed trypsin inhibitor and milk protein milk replacer were more efficient preweaning and overall than calves fed trypsin inhibitor and egg milk replacer. Results indicate that the blood derived colostrum supplement did not provide as much IgG as colostrum (4.55 g/L vs. 14.6 g/L, respectively), that feeding 1.0 g of trypsin inhibitor did not enhance serum IgG concentrations, and that the egg milk replacer-fed calves fed colostrum performed nearly as well as calves fed colostrum and the milk protein milk replacer.     

Effect of partial exchange of lactose with fat in milk replacer on ad libitum feed intake and performance in dairy calves

Compared with Holstein whole milk, commercial milk replacers (MR) for calves deliver relatively high levels of lactose and low levels of fat, and protein levels are rather comparable, resulting in a lower energy density and energy-to-protein ratio of the diet. Thus, the objective of this study was to determine the effects of partially exchanging lactose with fat in MR on voluntary feed intake, growth performance, and feeding behavior. Thirty-two male Holstein calves (2.1 ± 0.16 d of age, 46.4 ± 0.77 kg of body weight; BW) were assigned to 16 blocks of 2 calves per block based on arrival date and serum IgG. Within each block, calves were randomly assigned to 2 treatments: a high-lactose MR (HL; 17% fat; 44% lactose), or a high-fat MR (HF; 23% fat; 37% lactose). Lactose was exchanged by fat on a weight per weight basis, resulting in a 6% difference in metabolizable energy density per kilogram of MR. The experiment was divided into 3 phases: preweaning (P1; 0–35 d), weaning (P2; 36–56 d), and postweaning (P3; 57–84 d). For the first 2 wk of P1, calves were individually housed, fed their respective MR ad libitum through teat buckets, and provided access to water. At 14.2 ± 0.5 d of age, calves were group-housed (4 blocks/pen, 8 calves) and housed in group pens for the remainder of the study. In the group pens, calves were fed ad libitum MR, starter feed, chopped wheat straw, and water via automated feeders. During P2, calves were gradually weaned until complete milk withdrawal by 57 d and then monitored until 84 d (P3). Measurements included daily intakes and feeding behavior (rewarded and unrewarded visits), weekly BW and body measurements, and biweekly blood samples. Increasing fat content at the expense of lactose decreased MR intake during P1 by 15% (HL = 1.32 ± 0.04; HF = 1.17 ± 0.04 kg of dry matter per day), whereas total starter intake was not affected by MR composition. Once MR was restricted during P2, HL calves were reported to have more unrewarded visits to the automatic milk feeder than HF calves (11.9 ± 0.95 vs. 8.4 ± 1.03 visits/d, respectively). Crude protein intake was higher for HL calves during P1 (352.1 ± 11.2 vs. 319.6 ± 11.6 g/d), which was attributed to the higher intake of MR during that period, and metabolizable energy intake and protein-to-energy ratio remained comparable between treatments. Plasma cholesterol and nonesterified fatty acids levels were higher in HF calves as a consequence of the diet. Nevertheless, final BW (84 d) did not differ between treatments. Overall, calves fed ad libitum seemed to regulate their intake of MR based on its energy density, without significant effects on solid feed intake and overall growth.     

Comparison of a commercially available oral nutritional supplement and intravenous fluid therapy for dehydration in dairy calves

Calf scours is a primary cause of morbidity and mortality in the dairy industry. Effective treatments are needed to minimize death, maximize welfare, and maintain growth and productivity. The objective of this trial was to compare the efficacy of a commercially available nutritional supplement (Diaque, Boehringer-Ingelheim Vetmedica Inc., St. Joseph, MO) and i.v. lactated Ringer's solution (LRS) in rehydrating, preventing acidemia, and correcting electrolyte imbalances in an experimental model for calf scours. Twenty-four colostrum-fed suckling dairy calves were used in a modified crossover design. An osmotic diarrhea was induced by orally feeding commercial milk replacer modified with high level of sucrose to create a hypertonic milk solution, and administering oral hydrochlorothiazide and spironolactone for 48 h. The intention was to create a challenge sufficient to result in moderately dehydrated, standing calves without producing severe depression or loss of suckle. The efficacy of i.v. fluid therapy and a commercial nutritional supplement were subsequently compared for reversing the effects of the diarrheal disease. Treatment A consisted of administering the nutritional supplement according to label directions (100 g in 1.9 L of warm water, 3 times a day), and treatment B consisted of i.v. LRS (2 L, once a day). Clinical signs and laboratory results were obtained once daily by a blinded observer. The induction method was effective in creating the desired effect, as demonstrated by weight loss and subjective health and hydration scores. Both treatment groups experienced increases in body weight, base excess, and bicarbonate, and decreases in total protein and packed cell volume following treatment. Both i.v. LRS and Diaque are effective methods to correct hypovolemia and control derangements in acid-base status in calves with diarrhea and dehydration.     

Nitrogen utilization,preweaning nutrient digestibility,and growth effects of Holstein dairy calves fed 2 amounts of a moderately high protein or conventional milk replacer

Studies have shown that calves fed milk replacers (MR) with crude protein (CP) concentrations greater than 20%, as typically found in conventional MR, have higher dry matter intakes (DMI) and greater average daily gains (ADG) but consume less starter, which can lead to stress during weaning and reduced rumen development. The greater amount of CP being fed to preweaned calves may alter their nitrogen (N) balance, and excess N may be excreted in the urine. The objective of this study was to determine N utilization in preweaned calves fed diets varying in the amount of CP and MR fed. This study used 24 newborn dairy heifer calves blocked by birth and randomly assigned to 1 of 3 treatments: (1) 446 g dry matter (DM) of a conventional MR (CON; 20% CP, 20% fat), (2) 669 g DM of a moderately high protein MR (moderate; MOD; 26% CP, 18% fat), or (3) 892 g DM of a moderately high protein MR (aggressive; AGG; 26% CP, 18% fat). All calves had ad libitum access to starter and water. Both MR and starter were medicated with decoquinate. During weaning (d 43–49), the morning MR feeding ceased. On d 50, all MR feedings ended; however, starter and water intakes were continuously recorded until d 56. At 5 wk of age, urine was collected using urinary catheters for 3 d and chromium oxide was administered by bolus at 2 g/d for 7 d to estimate N efficiency. Calves fed MOD and AGG had similar starter intakes, feed efficiencies, and ADG, with the combined treatments having reduced starter intakes (258 vs. 537 g/d), greater ADG (674 vs. 422 g/d), and improved feed efficiency (0.57 vs. 0.45 gain:feed) compared with CON calves preweaning. However, DMI and water intake were similar across all treatments. Results from the N utilization phase showed that MOD and AGG treatments had similar but lower N efficiency compared with CON calves (45.5 vs. 52.7%). This could be due to MOD- and AGG-fed calves having greater urine volume and thereby, greater combined urine N output compared with CON calves (17.6 vs. 12.1 g/d). In summary, feeding >0.66 kg (DM) from a 26% CP MR increased ADG and improved feed efficiency during the preweaning period but reduced starter intake and lowered N efficiency.     

A multisite,randomized field trial to evaluate the influence of lactoferrin on the morbidity and mortality of dairy calves with diarrhea

Diarrhea is one of the most common causes of antimicrobial use and mortality in young calves. To reduce antimicrobial use and resistance on dairy farms, research on alternative therapies for calf diarrhea is necessary. Our laboratory previously conducted a randomized clinical trial investigating the effectiveness of lactoferrin, an iron-binding protein found in colostrum, as a treatment for calf diarrhea. The trial showed significantly reduced calf mortality in diarrheic calves that were administered lactoferrin. Thus, the objective of this study was to corroborate the results of our prior clinical trial across multiple farms and to investigate the effect of lactoferrin on the morbidity and mortality of preweaned calves with naturally occurring cases of diarrhea. This randomized field trial was conducted on 5 commercial dairy farms in Ohio. In total, 485 calves (≤21 d of age) were enrolled at first diarrhea diagnosis (fecal score ≥2 defined as loose to watery) and randomly assigned to receive an oral dose of lactoferrin (3 g of lactoferrin powder dissolved in 30 mL of water) or 30 mL of water (control) once daily for 3 consecutive days. Health assessments were conducted on the day of diarrhea diagnosis (d 0) and 1, 2, 3, 7, 14, 21, 28, and 35 d following diagnosis. Producer records of disease treatment and mortality were collected 120 d following diagnosis. A Poisson regression model was used to test differences between treatments in disease frequency through 35 d post-diarrhea diagnosis and the incidence risk of treatment and mortality risk 120 d post-diarrhea diagnosis; the model controlled for calf age at enrollment, farm, and treatment. Median calf age at enrollment was 11 d and ranged from 1 to 26 d of age. At study enrollment, 51.3% (123/240) and 52.2% (128/245) of calves in the control and lactoferrin treatment groups, respectively, were diagnosed with severe diarrhea (fecal score = 3). The frequency of disease (diarrhea, dehydration, depression, signs of respiratory disease) through 35 d following diarrhea diagnosis was not significantly different for calves in the lactoferrin and control groups. Overall mortality risk for enrolled calves was 9.9%, and 10.7% (22/243) and 9.1% (26/242) of calves in the lactoferrin and control groups, respectively, died or were culled in the 120 d following diarrhea diagnosis. The relative risk of death or culling did not differ between treatment groups, however. Therefore, as performed in this study, lactoferrin as a treatment for calf diarrhea was not beneficial.     

Effects of providing water from a bucket or a nipple on the performance and behavior of calves fed ad libitum volumes of acidified milk replacer

Little information is available on how much water ad libitum milk-fed calves consume during the milk-feeding stage and during weaning to solid feed. Further, no information exists on the effects on calf behavior and performance of different water delivery systems during these stages. Therefore, the objectives of this study were first, to examine the water intake of calves fed acidified milk replacer ad libitum during the milk-feeding and weaning stages, and second, to determine whether the method of water delivery affected water and feed intake and growth during these periods or oral behavior during weaning. To evaluate this, we registered feed and water intake, growth, and oral behaviors of 24 dairy calves before and after abrupt weaning from a 7-wk ad libitum acidified milk replacer feeding regimen. Two water sources (open bucket and nipple) were compared. During the 7-wk milk-feeding period, the calves drank, on average, 12.9 ± 0.7 L/d of acidified milk replacer but very little water. However, after abrupt removal of milk, the 2-mo-old calves rapidly increased their water intake. Moreover, no differences in water intake between the 2 water sources were noted either before or after abrupt weaning. Calves were observed to have some difficulties in using the water nipple. Calves provided water through the nipples consumed less water at each drinking bout but visited the water nipple more frequently compared with calves provided access to water from the bucket. We observed no differences in all other behaviors observed. Overall, the calves rapidly increased their feed intake, rumination time, frequency of cross-sucking, and vocalization after weaning. In conclusion, despite the calves’ consuming very little water when provided ad libitum access to acidified milk replacer, they dramatically increased their water consumption after abrupt weaning from milk. Calves did experience some difficulties in using the water nipples.     

Maternal late-gestation metabolic stress is associated with changes in immune and metabolic responses of dairy calves

Metabolic stress in periparturient dairy cows is characterized by excessive lipid mobilization, inflammation, and oxidative stress that is associated with immune dysfunction. Thus, metabolic stress around the time calving is linked to the development of various early-lactation health disorders. Maternal status during late pregnancy can have carryover effects on several health and production variables of neonatal calves. However, the effects of metabolic stress during gestation on metabolic and immune responses of newborn calves remain unknown. Thus, we aimed to investigate whether metabolic stress in late-gestation dairy cows is associated with changes in the metabolic and immune responses of their offspring during the first month of life. Holstein-Friesian cows (n = 12) were blood sampled at 28 and 15 d before expected calving. The average between these 2 sampling points in the serum concentrations of nonesterified fatty acids (NEFA), haptoglobin (Hp), and oxidant status index (OSi)—defined as the ratio between reactive oxygen and nitrogen species and total antioxidant potential—were calculated as indicators of the degree of lipid mobilization, inflammation, and oxidant status (OS), respectively. Calves were subsequently divided into groups (n = 6 each) according to their dams' high or low degree of lipid mobilization, inflammation, and OS. The metabolic responses of calves in each of these groups were compared weekly throughout their first month of life by assessing serum concentration of NEFA, Hp, and OSi. Additionally, whole blood was obtained from calves at each sampling period and subjected to a lipopolysaccharide (LPS)-stimulated tumor necrosis factor-α (TNF-α) production assay to assess cell-mediated innate immunity against induced inflammatory responses, using high (5 μg/mL of blood) and low (10 ng/mL) concentrations of LPS. Calves born to cows with higher NEFA or OSi showed lower body weight at birth and throughout the study, whereas no association between any of the maternal groups and average daily gain at 4 wk of age was identified. Serum concentrations of reactive oxygen and nitrogen species were higher in calves exposed to higher maternal NEFA concentrations or OSi when compared with calves born to cows with lower values of these biomarkers. Calves exposed to high maternal OS also had higher circulating concentrations of Hp and TNF-α, indicating greater basal inflammatory responses when compared with calves born to cows with a lower OSi. In contrast, LPS-induced inflammatory responses were less robust in calves exposed to higher maternal biomarkers of inflammation or OS, suggesting compromised immune responses to microbial agonists. Collectively, these data suggest that prenatal exposure to maternal parameters of metabolic stress may adversely affect some metabolic and inflammatory responses of the offspring that could influence disease susceptibility.     

Short communication: Effects of increasing protein and energy in the milk replacer with or without direct-fed microbial supplementation on growth and performance of preweaned Holstein calves

Forty-four Holstein calves were fed a direct-fed microbial (DFM) and 1 of 2 milk replacers to evaluate calf performance and growth. Treatments were (1) a control milk replacer [22:20; 22% crude protein (CP) and 20% fat], (2) an accelerated milk replacer (27:10; 27% CP and 10% fat), (3) the control milk replacer with added DFM (22:20+D), and (4) the accelerated milk replacer with added DFM (27:10+D). Dry matter intake, rectal temperatures, respiration scores and rates, and fecal scores were collected daily. Body weight, hip and withers height, heart girth, blood, and rumen fluid samples were collected weekly. Effects of treatment, sex, week, and their interactions were analyzed. Calves fed an accelerated milk replacer, regardless of DFM supplementation, consumed more CP and metabolizable energy in the milk replacer. No treatment differences were found for starter intake or intake of neutral detergent fiber or acid detergent fiber in the starter. Calves fed the accelerated milk replacer had greater preweaning and weaning body weight compared with calves fed the control milk replacer. Average daily gain was greater during the preweaning period for calves fed the accelerated milk replacer, but the same pattern did not hold true during the postweaning period. Feed efficiency did not differ among treatments. Hip height tended to be and withers height and heart girth were greater at weaning for calves fed the accelerated milk replacer compared with calves fed the control milk replacer. Fecal scores were greatest in calves fed DFM. Overall acetate, propionate, butyrate, and n-valerate concentrations were lower in calves fed the accelerated milk replacer, but DFM did not have an effect. Rumen pH was not different. Blood metabolites were unaffected by DFM supplementation, but calves fed the accelerated milk replacer had increased partial pressure of CO₂, bicarbonate, and total bicarbonate in the blood. Direct-fed microbial supplementation did not appear to benefit the calf in this trial     

Reducing gut effects from Cryptosporidium parvum infection in dairy calves through prophylactic glucagon-like peptide 2 therapy or feeding of an artificial sweetener

Glucagon-like peptide 2 (GLP-2) therapy was shown previously to reduce inflammation-related gut damage from coccidiosis in dairy calves, and feeding of artificial sweetener stimulates GLP-2 secretion from intestinal L cells. The purpose of this study was to determine whether GLP-2 treatment or artificial sweetener feeding beginning 1 wk before an experimental inoculation with the coccidian parasite Cryptosporidium parvum can reduce infection-related intestinal damage in Holstein bull calves. Newborn calves were assigned to 1 of 4 treatment groups of 6 calves each, including noninfected control calves injected s.c. every 12 h with control buffer (CON), infected control calves injected s.c. every 12 h with control buffer (INF), infected calves injected s.c. every 12 h with 50 µg/kg of body weight of GLP-2 (GLP2), and infected calves injected s.c. every 12 h with control buffer and supplemented in the diet with Sucram (Pancosma, Geneva, Switzerland) at 400 mg/kg of dry matter of milk replacer (SUC). Treatments were initiated on d 1, and calves in INF, GLP2, and SUC were orally dosed on d 8 with 12,500 C. parvum oocysts. Fecal scores were recorded daily, plasma was collected on d 1, 8, 12, 15, and 18 to evaluate markers of inflammation, and fecal samples were collected on d 1, 8, and every other day thereafter to determine the presence of oocysts. Calves were euthanized on d 18 for collection of intestinal tissues and histological and gene expression analyses. Relative to CON, calves in INF exhibited an increase in diarrhea severity, increased plasma serum amyloid A concentration on d 15 and 18, reduced intestinal villus height, increased villus apoptosis and crypt cell proliferation, and increased intestinal mRNA expression of MARVELD2 and GPX2. However, calves in SUC and GLP2 had reduced diarrhea severity and fecal C. parvum oocyst shedding, reduced plasma serum amyloid A concentration on d 15 and 18, and, depending on the intestinal segment, increased villus height, reduced crypt cell proliferation, and reduced mRNA expression of MARVELD2, GPX2, and other tight junction proteins relative to INF. Lastly, GLP2 and SUC exhibited increased intestinal mass-to-length ratio and decreased length-to-empty body weight ratio relative to INF. Our findings suggest that GLP-2 and Sucram treatments administered before a low-level C. parvum exposure may contribute to fewer effects on intestinal integrity, morphology, and inflammation in response to infection, and shorter, denser intestines.     

Effects of feeding unlimited amounts of milk replacer for the first 5 weeks of age on rumen and small intestinal growth and development in dairy calves

The development of the gastrointestinal tract in newborn calves is essential for sufficient nutrient uptake. An intensive milk feeding during the neonatal period may impair the rumen development in calves. The aim of this study was to investigate effects of milk replacer (MR) feeding in unlimited amounts for the first 5 wk of age on the gastrointestinal growth and development in preruminant calves at wk 9 of age. Twenty-eight newborn Holstein and Holstein × Charolais crossbred calves (19 male and 9 female) were fed MR ad libitum (ADLIB) or in restricted amounts (6 L per day; RES) until wk 5 of age. Thereafter, the MR intake of ADLIB was gradually reduced at wk 6 and 7, and all calves received 6 L of MR per day until wk 9 of age. In wk 9, calves were slaughtered and carcass and organ weight as well as rumen papilla size in the atrium, ventral sac, and ventral blind sac, and villus size of the mucosa in the small intestine (duodenum; proximal, mid, and distal jejunum; and ileum) were determined. The expression of mRNA associated with the local insulin-like growth factor (IGF) system was measured in the rumen epithelium. Ad libitum MR feeding increased MR intake and growth in ADLIB without influencing concentrate intake compared with RES. Carcass weight in wk 9 was greater in ADLIB than in RES. The density of the rumen papillae in the atrium and ventral blind sac was greater in RES than in ADLIB calves, but surface area of the epithelium was not different between groups in the investigated regions of the rumen. The mRNA abundance of IGF1 in the atrium tended to be greater and the IGFR1 mRNA abundance in the ventral sac tended to be lower in the ADLIB than in the RES feeding group. The rumen pH and volatile fatty acid concentrations were not affected by MR feeding intensity. In mid-jejunum, villus circumference was greater in ADLIB than in RES calves. In the distal jejunum, villus surface area and the villus height/crypt depth ratio were greater and the villus circumference and height tended to be greater, whereas crypt depth was smaller in ADLIB than in RES calves. The findings from this study indicate that ad libitum MR feeding for 5 wk of age followed by its gradual reduction promotes growth performance without any negative influence on gastrointestinal growth and development in dairy calves at 9 wk of age.