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.     

Exposure to antimicrobials through the milk diet or systemic therapy is associated with a transient increase in antimicrobial resistance in fecal Escherichia coli of dairy calves

The objective of this prospective cohort study was to describe the relationship between exposure to antimicrobials, through both the milk diet and systemic therapy, and to describe antimicrobial resistance of fecal Escherichia coli in dairy calves pre- and postweaning. A convenience sample of 15 Minnesota dairy farms was chosen, representing 3 equal cohorts of milk diet fed to preweaned calves: medicated milk replacer (MMR), nonmedicated milk replacer (NMR), or pasteurized nonsaleable milk (PNM). Five newborn calves were enrolled on each farm, with fecal samples collected from each calf at 1, 3, 5, and 16 wk of age. After isolation, 3 colonies of E. coli were randomly selected from each sample to determine antimicrobial susceptibility by minimum inhibitory concentration (Sensititer, Thermo Scientific, Waltham, MA) to 8 antimicrobials in 8 classes. The isolate was given an antimicrobial resistance score (ARS) according to the number of antimicrobial classes to which it was resistant. Any isolate resistant to 3 or more antimicrobials was defined as being multidrug resistant (MDR). Relationships between ARS and MDR (dependent variables) and possible explanatory variables were analyzed using mixed multivariable linear and logistic regression models, respectively, with critical P-values adjusted for multiple contrasts. Seventy percent of isolates were resistant to sulfadimethoxine. For wk 1 and 3, the mean ARS values were greatest for fecal E. coli from calves fed MMR or PNM compared with NMR, with no difference in ARS values between the MMR and PNM groups at either time point. At wk 5, the mean ARS value was greatest for fecal E. coli from calves fed MMR (3.56 ± 0.45; mean ± SE), intermediate for calves fed PNM (2.64 ± 0.45), and lowest for calves fed NMR (1.54 ± 0.45). However, by wk 16, the mean ARS values were ≤1.0 and did not differ among milk diets. Evaluation of the proportion of isolates with MDR mirrored the results of the ARS analysis (MDR more prevalent in MMR and PNM groups preweaning; no difference among milk diets at 16 wk). There was a tendency for an increase in ARS at wk 5 (1.28 ± 0.70), and the odds for MDR in fecal E. coli were estimated to be 5.2 (95% confidence interval = 0.67, 35.7) and 101.1 (95% confidence interval = 1.15, >999.9) higher at wk 3 and 5 if the calf was treated with a systemic antimicrobial within the 14-d period before sampling. These findings suggest that exposure to antimicrobials through the milk diet or systemic therapy may result in a transient increase in resistance in fecal E. coli, but once the antimicrobial pressure is removed, susceptible E. coli are able to flourish again, resulting in an overall decrease in resistance.     

Prevalence,antimicrobial susceptibility,and antibiotic resistance gene transfer of Bacillus strains isolated from pasteurized milk

Pasteurization is carried out in dairy industries to kill harmful bacteria present in raw milk. However, endospore-forming bacteria, such as Bacillus, cannot be completely eliminated by pasteurization. In this study, a total of 114 Bacillus strains were isolated from 133 pasteurized milk samples. Antibiotic susceptibility tests showed that the percentage of Bacillus with intrinsic resistance to ampicillin and penicillin were 80 and 86%, respectively. Meanwhile, some Bacillus isolates had acquired resistance, including trimethoprim-sulfamethoxazole resistance (10 isolates), clindamycin resistance (8 isolates), erythromycin resistance (2 isolates), and tetracycline resistance (1 isolate). To further locate these acquired resistance genes, the plasmids were investigated in these 16 Bacillus strains. The plasmid profile indicated that Bacillus cereus BA008, BA117, and BA119 harbored plasmids, respectively. Subsequently, the Illumina Novaseq PE150 was applied for the genomic and plasmid DNA sequencing. Notably, the gene tetL encoding tetracycline efflux protein was found to be located on plasmid pBC46-TL of B. cereus BA117. In vitro conjugative transfer indicated that pBC46-TL can be transferred into Bacillus invictae BA142, Bacillus safensis BA143, and Bacillus licheniformis BA130. The frequencies were of 1.5 × 10^?7 to 1.7 × 10^?5 transconjugants per donor cells. Therefore, Bacillus strains with acquired antibiotic resistance may represent a potential risk for the spread of antibiotic resistance between Bacillus and other clinical pathogens via horizontal gene transfer.     

Effect of waste milk pasteurization on fecal shedding of Salmonella in preweaned calves

The objective of the current research was to determine if pasteurization of nonsaleable waste milk influences fecal Salmonella concentrations and prevalence, or antimicrobial susceptibility and serotype of the cultured isolates. Holstein dairy calves (n = 211) were housed on a single commercial dairy in the southwestern United States and randomly allotted to be fed either pasteurized (PWM; n = 128 calves) or nonpasteurized waste milk (NPWM; n = 83 calves). Fecal samples were collected via rectal palpation or from freshly voided, undisturbed fecal pats, weekly during the first 4 wk of the animal's life and then again at weaning. Eight total collections were made and 1,117 fecal samples cultured for Salmonella. One isolate from each culture-positive fecal sample was preserved for antimicrobial susceptibility screening and serotyping. Sixty-nine percent of the fecal samples were culture positive for Salmonella with no difference due to treatment (67.7 and 69% Salmonella positive for PWM and NPWM treatments, respectively). Few fecal samples (178/1,117; 15.9%) contained Salmonella concentrations above the limit of detection (～1 cfu/g of feces) with concentrations ranging from 1.0 to 6.46 cfu (log₁₀)/g of feces. Concentration was not affected by treatment. Seventeen different serotypes were identified, the majority of which were Montevideo and Anatum. A greater percentage of Typhimurium (87 vs. 13%), Muenchen (88 vs. 12%), and Derby (91 vs. 9%) were recovered from calves fed PWM compared with NPWM-fed calves. Conversely, Newport (12.5 vs. 86%), Bredeney (22.2 vs. 77.8%), and Muenster (12.5 vs. 87.5%) were lower in PWM compared with NPWM treatments. The majority (66.7%) of isolates were susceptible to all of the antibiotics examined. Results from this one commercial dairy suggest that milkborne Salmonella is not an important vector of transmission in dairy neonates, nor does pasteurization of waste milk influence fecal shedding of this pathogen. Caution should be used, however, when extrapolating results to other farms as Salmonella contamination of milk on farm is well documented. The potential benefits of pasteurization in disease prevention outweigh the potential risks of feeding a nonpasteurized product and warrants incorporation into any calf-rearing program using nonsaleable waste milk for feeding young dairy neonates.     

Survey on the disposal of waste milk containing antimicrobial residues on Swiss dairy farms

Feeding waste milk containing antimicrobial residues (WMA) to calves has been associated with increased antimicrobial resistance in calves' commensal bacterial flora. The objectives of this study were (1) to document practices related to the disposal of WMA on Swiss dairy farms and (2) to evaluate the association between farm characteristics and the feeding of WMA to calves. A web-based questionnaire on practices surrounding waste milk disposal was completed by 1,625 dairy producers (10.9% of solicited producers). Logistic regression models were built to evaluate the association between herd characteristics and the practice of feeding WMA. Waste milk produced during and up to the first milking after completion of antimicrobial treatment or during the withdrawal period was fed to at least some of the calves on 47.3% of respondents' farms. Farms in organic production had lower odds of feeding WMA to calves than nonorganic farms [odds ratio (OR) 0.59]. Farms located in the eastern region of Switzerland, as opposed to those in the western region, had increased odds of feeding WMA to calves (OR 2.01). A yearly average bulk tank somatic cell count ≥150,000 cells/mL was associated with increased odds of feeding WMA to calves compared with the reference category of <100,000 cells/mL (OR 1.62). An average cow-level annual milk production ≥8,500 L was associated with increased odds of feeding WMA to calves compared with farms in the interquartile range with a production of 6,500 to 8,499 L (OR 1.24). Further research is warranted to investigate dairy farmers' motivations affecting this practice, and to quantitatively define calves' exposure to antimicrobial residues and the resulting antimicrobial resistance in calves' commensal flora on these farms.     

Multidrug residues and antimicrobial resistance patterns in waste milk from dairy farms in Central California

Waste milk (WM) is a common source of feed for preweaned calves in US dairy farms. However, limited information is available about characteristics of this product, including concentration of drug residues and potential hazards from antibiotic-resistant bacteria present in the milk. The aims of this cross-sectional study were to (1) identify and measure the concentration of antimicrobial residues in raw WM samples on dairy farms in the Central Valley of California, (2) survey farm management practices for factors associated with the occurrence of specific antimicrobial residues in raw WM, (3) characterize the antimicrobial resistance patterns of E. coli cultured from raw WM samples, and (4) evaluate the potential association between WM quality parameter and risk of identifying drug residues in milk. A single raw bulk tank WM sample was collected from dairy farms located in California's Central Valley (n = 25). A questionnaire was used to collect information about farm management practices. Waste milk samples were analyzed for a multidrug residue panel using liquid chromatography–tandem mass spectrometry. Bacteria were cultured and antimicrobial resistance was tested using standard techniques; milk quality parameters (fat, protein, lactose, solids-not-fat, somatic cell count, coliform count, and standard plate count) were also measured. Of the 25 samples collected, 15 (60%) contained detectable concentrations of at least 1 antimicrobial. Of the drug residue–positive samples, 44% (11/25) and 16% (4/25) had detectable concentrations of β-lactams and tetracycline, respectively. The most prevalent drug residues were ceftiofur (n = 7, 28%), oxytetracycline (n = 4, 16%), and cephapirin (n = 3, 12%). No significant associations were identified between farm characteristics or management practices and presence of drug residues in WM. In this study, 20% of farms did not pasteurize WM before feeding to calves. Two of the 10 Escherichia coli isolated from WM samples were multidrug resistant. Streptococcus spp. (n = 21, 84%) was the most common genus cultured from WM samples, followed by Staphylococcus spp. (n = 20, 80%) and E.coli (n = 10, 40%). Mycoplasma spp. was cultured from 2 WM samples (n = 2, 8%). The presence of drug residues in WM at concentrations that increase selection of resistant bacteria indicates the need for additional studies targeting on-farm milk treatments to degrade drug residues before feeding to calves. The presence of multidrug-resistant E. coli in WM urges the need for on-farm practices that reduce calf exposure to resistant bacteria, such as pasteurization.     

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.     

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.     

Tributyrin supplementation in pasteurized waste milk: Effects on growth performance,health, and blood parameters of dairy calves

This study evaluated the effects of incremental tributyrin supplementation in pasteurized waste milk on growth performance, health, and blood metabolism of dairy calves before and after weaning. Forty-eight newborn female Holstein dairy calves (39.6 ± 2.75 kg; mean ± standard deviation) were blocked by age and randomly assigned to 3 treatments: pasteurized waste milk (1) without supplementation, (2) with 1 g/L of tributyrin products (unprotected solid powder; containing 35% tributyrin), or (3) with 2 g/L of tributyrin products. The calves were weaned on d 56 and were raised until d 77. Data were analyzed for the preweaning, postweaning, and overall periods. The results showed that starter intake and hay intake were not different among treatments in any period of the trial, but the crude protein intake tended to increase linearly with tributyrin supplementation during the overall period. Although tributyrin supplementation had no effects on body weight during preweaning and overall periods, body weight increased linearly with tributyrin supplementation postweaning. The average daily gain tended to increase linearly during postweaning and overall periods. No effects were observed on feed efficiency in any period. A positive linear relationship between body length and tributyrin supplementation was observed during the postweaning period, but no differences were found for the other body structural measurements in any period. The results of diarrhea showed that tributyrin concentration had a negative linear relationship with diarrhea frequency during preweaning and overall periods. The rectal temperature did not differ among treatments in any period, but a treatment × week effect for rectal body temperature was observed. For blood metabolism, tributyrin supplementation had no effects on insulin, growth hormone, total protein, albumin, or globulin. No differences were found in serum amyloid A concentration in any of the periods, yet haptoglobin concentration decreased linearly with increasing tributyrin concentration during postweaning and overall periods. Endothelin concentration showed a tendency to decrease linearly during preweaning and postweaning periods and decreased linearly with tributyrin supplementation during the overall period. An increasing tributyrin concentration was associated with a negative linear relationship with IL-1β concentration during the preweaning period, and no differences were found in the other periods. The concentration of IL-6 and tumor necrosis factor α were not different among treatments in any of the periods. These data suggest that increasing the concentration of tributyrin in pasteurized waste milk could increase growth performance and health of dairy calves, and incremental tributyrin supplementation could linearly reduce haptoglobin, endothelin, and IL-1β concentrations, indicating a positive effect of tributyrin on alleviating oxidative stress and inflammatory status of dairy calves. Calves fed pasteurized waste milk supplemented with tributyrin products (containing 35% tributyrin) at 2 g/L compared with 1 g/L of milk had more improved growth and health.     

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.     

The effects of milk feeding method and group size on feeding behavior and cross-sucking in group-housed dairy calves

This study investigated the effect of teat feeding and group size on cross-sucking and competition for milk in dairy calves. Ninety-six Holstein-Friesian male and female calves were allocated to either pairs or groups of 6 and fed milk either from a bucket or via a teat. Calves fed via a teat spent more time ingesting the milk. They spent time sucking the empty teat after milk intake and they spent less time cross-sucking compared with calves fed from buckets. The results show that teat feeding reduces cross-sucking in groups of 6 calves. Calves in groups of 6 ingested the milk faster than calves housed in pairs, which suggests that the competition for milk was greater than in pairs. Teat-fed calves changed to another teat more often than bucket-fed calves changed to another bucket during milk intake. Thus, the use of teat feeding did not reduce the competition for milk as compared with bucket feeding and future studies should focus on improving teat-feeding methods so that they reduce competition for milk in group-housed calves.     

生猪屠宰环节沙门氏菌携带状况、耐药性及其耐药基因分析

目的 调查河南省生猪屠宰环节沙门氏菌污染情况, 并分析其耐药性及耐药基因。方法 本研究从河南省不同规模的生猪屠宰企业共采集825份样品, 通过常规检测方法和分子生物学技术对样品进行沙门氏菌分离鉴定, 然后通过药敏实验检测沙门氏菌分离株对17种抗菌药的敏感性; 对部分沙门氏菌分离株进行了12种耐药基因检测。结果 样品中共分离出沙门氏菌241株, 总分离率为29.21%; 所有受试沙门氏菌分离株均对阿米卡星敏感, 而对多西环素耐受性最强(耐药率为95.00%), 其次对其他抗菌药都表现出不同程度的耐受性; 上述分离株存在着严重的多重耐药问题, 其中耐3种及以上药物的菌株占比高达98.34%。耐药基因检测结果显示, 受试菌株均含有针对全部5大类受试药物的抗性基因, 但aac(3)-Ⅰa、gurA、tetB 3个耐药基因均未有检出。结论 河南省生猪屠宰企业存在严重的沙门氏菌污染情况, 且菌株存在严重的耐药性。     

Effects of once- versus twice-a-day feeding of pasteurized milk supplemented with yeast-derived feed additives on growth and health in female dairy calves

The objective of this study was to compare the effects of feeding preweaning dairy calves pasteurized milk once or twice a day with or without a combination of yeast-derived products. Holstein heifer calves (n = 48) from The Pennsylvania State University dairy herd were fed 3.8 L of colostrum in 1 feeding and randomly assigned to 1 of 4 treatments (once-a-day milk feeding with or without live yeast culture and mannan-rich fraction and twice-a-day milk feeding with or without live yeast culture and mannan-rich fraction). All calves were fed 6 L of milk daily. Weekly growth measurements and blood samples were taken 3 h after the morning milk feeding for all animals. Growth measurement included body weight, hip width, and withers height. Calf starter refusal was recorded weekly, and a sample was taken to determine dry matter intake. Daily health scores were recorded for each calf using a standard scoring system. Intake, growth measurements, haptoglobin, and health scores data were analyzed using repeated measures analysis with calf included as a random variable. Preweaning average daily gain was 553.4 and 512.1 g/d for calves fed milk once and twice a day, respectively, and we found no difference between treatments. Preweaning calf starter intake was 242.3 and 198.7 g/d for calves fed milk once and twice a day, respectively, and we found no treatment differences. Preweaning calf starter intake was 224.3 and 216.6 g/d for calves fed yeast and without yeast, respectively. Withers height and hip width were similar in calves fed milk either once or twice a day; however, calves fed yeast tended to have greater withers height and hip width than control calves. Haptoglobin concentration as a measure of stress had least squares means of 4.0 and 9.5 ± 3.5 µg/mL for calves fed milk once or twice a day, respectively, and we found no difference among treatments. Scours score and total daily score were similar for calves fed milk once or twice a day. These results suggest that feeding milk once a day can be successfully applied to a calf feeding system and that yeast products may improve structural growth.     

Effects of spray-dried animal plasma in calf milk replacer on health and growth of dairy calves

Male Holstein calves (n = 120) purchased from local dairy farms were fed one of three calf milk replacers for 42 d. Experimental milk replacers were formulated to contain whey protein concentrate (WPC) as the primary protein source or WPC plus 5% spray-dried bovine plasma (SDBP) or spray-dried porcine plasma (SDPP). The SDPP was heated to remove heat-insoluble materials and provide products with similar IgG content. Calves were also fed commercial calf starter and water for ad libitum consumption. Intake, change in body weight (BW), feed efficiency, morbidity and mortality were determined. Mortality was 10, 3, and 2 in calves fed WPC, SDBP, and SDPP treatments, respectively. Morbidity, measured as the number of days that calves had diarrhea was reduced by 30% when SDBP or SDPP were fed. Calves had diarrhea for 6.9, 3.9, and 4.7 d during the 42-d study when fed commercial calf milk replacer containing WPC, SDBP, and SDPP, respectively. Fecal scores tended to be reduced and feed efficiency tended to be improved when SDBP or SDPP were fed. Mean intakes of total dry matter during the 42-d study were greater when calves were fed SDBP or SDPP and were 661, 710, and 684 g/d for calves fed WPC, SDBP, and SDPP, respectively. Mean BW gains from d0 to 42 were 231,261, and 218 g/d, respectively. Calves fed SDPP tended to have lower BW gain during the first 28 d of the study. However, difference in daily BW gain from d 1 to 28 was only 39 g/d. Inclusion of SDBP or SDPP in milk replacer reduced morbidity and mortality of milk-fed dairy calves.     

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.     

Effects of preweaning total plane of milk intake and weaning age on intake,growth performance,and blood metabolites of dairy calves

The objective of this study was to evaluate the effects of preweaning total plane of milk intake and weaning age on intake, growth performance, and blood metabolites of dairy calves. A total of 48 Holstein calves (40 ± 1.6 kg of body weight) were used in a 2 × 2 factorial arrangement with the factors of weaning age (d 60 vs. 75) and the total plane of milk intake (medium vs. high) during the preweaning period. Calves were assigned to 1 of 4 treatments: (1) calves fed medium plane of milk (MPM) intake and weaned on d 60 of age (MPM-60d, 4 L/d of milk from d 3 to 10, 6 L/d of milk from d 11 to 55, and 3 L/d of milk from d 56 to 60 of age; total milk intake = 317 L), (2) calves fed MPM intake and weaned on d 75 of age (MPM-75d, 4 L/d of milk from d 3 to 10 and 4.5 L/d of milk from d 11 to 70 of age followed by feeding 2.25 L/d of milk from d 71 to 75 of age; total milk intake = 313 L), (3) calves fed high plane of milk (HPM) intake and weaned on d 60 of age (HPM-60d, 4 L/d of milk from d 3 to 10, 6 L/d of milk from d 11 to 20, and 8.5 L/d of milk from d 21 to 55 followed by feeding 4.25 L/d of milk from d 56 to 60 of age; total milk intake = ～411 L); and (4) calves fed HPM intake and weaned on d 75 (HPM-75d, 4 L/d of milk from d 3 to 10, and 6 L/d of milk from d 11 to 70 of age followed by feeding 3 L/d of milk from d 71 to 75 of age; total milk intake = 407 L) with no milk refusals. All of the calves were monitored up to d 90 of age. Regardless of weaning age, starter feed intake and dry matter intake (% of body weight) were lower in calves fed HPM compared with those receiving MPM. A tendency for the plane of milk intake × weaning age interaction was observed for metabolizable energy intake with the highest value was recorded with the HPM-75d calves. The lowest efficiency of metabolizable energy intake and average feed efficiency was observed in HPM-60d calves throughout the experimental period as compared with the other groups. An interaction was found between the total plane of milk intake and weaning age regarding effects on total average daily gain, average daily gain/metabolizable energy intake, feed efficiency, final body weight, and plasma β-hydroxybutyrate levels with the highest values measured in HPM-75d calves. Weaning on d 75 versus d 60 improved wither height and hip width, which tended to increase body length at the end of the trial. The results suggest that calves fed high amounts of milk during their preweaning period benefit from extending the time of weaning from 60 to 75 d of age based on average daily gain, feed efficiency, and final body weight.     

Effects of feeding different amounts of milk replacer on growth performance and nutrient digestibility in Holstein calves to 2 months of age using different weaning strategies

Growth and the digestibility of nutrients can be greatly affected by diet preweaning and the rate at which calves are weaned. A 2 × 2 factorial design [moderate (MOD) or high (HI) milk replacer (MR) feeding rates and abrupt (AB) or gradual (GR) weaning] was used to compare these effects. Calves (n = 50) were randomly assigned to 1 of 4 treatments: MOD-AB, MOD-GR, HI-AB, and HI-GR. Calves assigned to MOD-AB were fed 0.66 kg of MR for the first 42 d and then 0.33 kg for the last 7 d; those assigned to MOD-GR were fed 0.66 kg of MR for 28 d, 0.33 kg for 14 d, and 0.17 kg for the last 7 d; those assigned to HI-AB were fed 0.66 kg of MR for 7 d, 0.82 kg for 7 d, 1.1 kg for 28 d, and 0.66 kg for the last 7 d; and those assigned to HI-GR were fed 0.66 kg of MR for 7 d, 0.82 kg for 7 d, 1.1 kg for 14 d, 0.66 kg for 14 d, and 0.33 kg for the last 7 d. All calves received the same MR [25% crude protein (CP), 17% fat; dry matter (DM) basis] and were given ad libitum access to water and a textured starter (42% starch and 20% CP). On d 26 to 30 and d 45 to 49, a fecal sample was taken from 5 calves in each treatment via the rectum to estimate apparent digestibility coefficients (dC). Apparent dC of DM, organic matter, and fat were greater for HI versus MOD calves. Apparent dC of neutral detergent fiber, acid detergent fiber, and sugar were greater for MOD versus HI calves. Apparent dC of DM was greater for AB versus GR calves [90.9, 89.0 ± 0.5384 (standard error)], and the apparent dC of acid detergent fiber was greater for GR versus AB calves (39.5, 32.3 ± 1.67). Feed efficiency was greater for HI versus MOD and AB versus GR. There were no significant differences between CP or starch dC based on treatment, and no interactions were observed. Starter consumption was greater for MOD calves compared with HI calves, and GR calves consumed more than AB calves. These results suggest that providing calves high amounts of MR preweaning enhanced readily available nutrient dC, but providing moderate amounts of MR resulted in increased fibrous fraction dC. Accordingly, the AB weaning strategy had higher dC for DM and organic matter, but there was a depression in fiber dC.     

Effect of milk replacer program on digestion of nutrients in dairy calves

There are concerns with feeding young dairy calves amounts of milk solids approaching 0.9 kg of dry matter (DM) or more because of slumps in average daily gain (ADG) at weaning and low starter intakes. Additionally, programs feeding more than 0.6 kg of DM have not been thoroughly tested for success at different weaning ages. Four milk replacer (MR) programs were compared in trial 1. Program A was 0.44 kg of DM of a 21% crude protein (CP), 21% fat MR powder fed daily for 42 d. Program B was 0.66 kg of DM of a 27% CP, 17% fat MR powder fed daily for 42 d. Program C was 0.66 kg of DM of a 27% CP, 17% fat MR powder daily fed for 28 d. Program D was up to 1.09 kg of DM of a 29% CP, 21% fat MR daily fed for 49 d. Digestibility estimates were made and blood was sampled for serum constituents on d 53 to 56, and performance was measured for 84 d. Three programs feeding a 27% CP, 17% fat MR powder were compared in trial 2 over 56 d. Calves on program A were fed 0.66 kg of DM powder daily and weaned at 28 d. Calves on program B were fed 0.66 kg of DM powder daily and weaned at 42 d. Calves on program C were fed up to 1.09 kg of DM powder daily and weaned at 42 d. Digestibility estimates were made and blood was sampled for serum constituents d 21 to 24, d 36 to 39, and d 53 to 56. In trial 1, calves fed program A had the least overall ADG. Calves fed program D had the greatest ADG from 0 to 56 d, the least ADG from d 56 to 84, the least digestibility estimates, and the least concentrations of serum amylase. At 84 d, there were no differences in body weights of calves fed programs B, C, and D. In trial 2, calves fed program A had the greatest starter intake and greatest concentrations of serum amylase. Calves fed program C had the least estimates of digestibility from d 53 to 56 and the least serum concentrations of amylase. Calves fed up to 1.09 kg/d of 27 to 29% MR powders and weaned at 42 or 49 d had lower starter intakes, concentrations of serum amylase, and digestion of starter postweaning compared with calves fed conventional 21% CP, 21% fat MR powders fed at 0.44 kg/d. Calves fed 0.66 kg/d of a 27% CP, 17% fat MR powder and weaned at 28 or 42 d of age had no reductions in intake or digestion compared with calves fed conventional MR and gained as much total body weight from 0 to 84 d as calves stepped up to 1.09 kg of MR.     

Baicalin inhibits Escherichia coli isolates in bovine mastitic milk and reduces antimicrobial resistance

In this study, we aimed to evaluate the inhibitory effect of baicalin on Escherichia coli in vitro and the effects of baicalin treatment on antimicrobial resistance of the E. coli isolates. Through isolation, purification, and identification, a total of 56 E. coli strains were isolated from 341 mastitic milk samples. The study of inhibition effect of baicalin on the E. coli strains in vitro was focused on permeability and morphology of the isolates using an alkaline phosphatase kit and scanning electron microscopy. Furthermore, the resistance spectrum of the isolates to the common antimicrobial agents was tested at sub-minimum inhibitory concentrations of baicalin by the agar dilution method. Extended-spectrum β-lactamase and plasmid-mediated quinolone resistance genes were amplified by PCR before and after incubation with baicalin. The results revealed that baicalin has certain inhibitory effects on the isolates in vitro. The alkaline phosphatase enzyme activity was significantly increased from 1.246 to 2.377 U/100 mL, and the surface of E. coli was concave and shriveled. Analysis of the resistance spectrum and PCR amplification showed that, after administration with baicalin, the sensitivity of most strains to the selected antimicrobial agents was enhanced. Strikingly, the drug-resistant genes from 71.43% (40/56) of these isolates were found to have drug-resistant genes to different extents. Altogether, the current study confirmed both the inhibitory effect on Escherichia coli in vitro and the reduction of antimicrobial resistance by baicalin. This is the first comprehensive study to report on baicalin, a traditional Chinese medicine that acts on E. coli isolated from the mastitic milk samples.     

Effect of feeding frequency and route of administration on abomasal luminal pH in dairy calves fed milk replacer

The aim of this study was to determine the effect of feeding frequency and route of administration on abomasal luminal pH in suckling calves. Six male dairy calves with cannulae in the abomasal body were administered the following six treatments in a randomized crossover design: 24 h fasting, suckling of a high-quality milk replacer (all-milk protein; 12% of body weight [BW]/d) at 12-h (2x), 8-h (3x), 6-h (4x), and 3-h (8x) intervals, and ruminal intubation of milk replacer (12% of body weight/day) at a 12-h (2x) interval. Abomasal luminal pH was measured every second for 24 h with miniature glass pH electrodes. Least squares mean 24-h fasting abomasal luminal pH was 1.73, whereas mean 24-h pH after suckling and intubation of milk replacer every 12 h were higher at 3.44 and 3.17, respectively. Increasing the frequency of milk replacer suckling to 3x, 4x, and 8x increased mean 24-h abomasal luminal pH; however, there was no difference in mean 24-h pH between 3x (3.69), 4x (3.64), and 8x (3.67) suckling. The percentage of the 24-h recording period that abomasal luminal pH was > 3.0 was 0, 49, 53, 61, 61, and 71% for fasting, 2x intubation of milk replacer, and 2x, 3x, 4x, and 8x suckling of milk replacer, respectively. Increasing the frequency of milk replacer suckling may be efficacious in the prophylaxis of abomasal ulceration in milk-fed calves.