A randomized trial to study the effect of automatic cluster remover settings on milking performance, teat condition, and udder health.

The objectives were to study the effect of 2 different automatic cluster remover settings on (1) milking characteristics, (2) milk component yields, (3) teat tissue condition, and (4) udder health. In a randomized controlled field trial, Holstein cows (n = 689) from 1 commercial dairy farm with a thrice-daily milking schedule were allocated to 2 treatment groups. Treatment consisted of a cluster remover take-off milk flow threshold of 1.2 (ACR1.2) or 0.8 kg/min (ACR0.8) for 57 d. Milking characteristics (milk yield; and milking unit-on time, MUOT) were obtained with electronic on-farm milk meters. Composite milk samples were collected and analyzed for fat, protein, lactose, and somatic cell count. Machine-milking-induced short- and long-term changes to the teat tissue condition were assessed visually. General linear mixed models demonstrated differences in MUOT, whereas no meaningful differences in milk yield were detected. Milk yield (least squares means, 95% confidence interval) was 11.3 (10.9-11.8) and 11.3 (10.8-11.8) kg in groups ACR1.2 and ACR0.8, respectively. The effect of treatment on MUOT was modified by parity. Milking unit-on time in first-, second-, and ≥third-lactation cows, respectively, was 260.7 (252.0-269.4), 257.8 (247.4-268.1), and 260.2 (252.6-267.9) s in group ACR1.2; and 273.7 (264.9-282.5), 279.1 (269.4-288.8), and 295.7 (287.9-303.6) s in group ACR0.8. We detected no meaningful differences in milk component yields or linear somatic cell score. Least squares means in groups ACR1.2 and ACR0.8, respectively, were milk fat yield, 0.42 (0.40-0.44) and 0.42 (0.40-0.44) kg; milk protein yield, 0.36 (0.35-0.37) and 0.37 (0.36-0.37) kg; milk lactose yield, 0.61 (0.60-0.63) and 0.63 (0.61-0.64) kg, and linear somatic cell score, 1.9 (1.8-2.0) and 1.9 (1.8-2.0). A generalized linear mixed model revealed an effect of treatment on machine-milking-induced short-term changes. The odds of short-term changes to the teat tissue were lower for cows in group ACR1.2 [odds ratio (95% confidence interval) = 0.78 (0.63-0.96)]. No meaningful differences were detected in machine-milking-induced long-term changes between treatment groups. Increasing cluster remover take-off milk flow threshold from 0.8 to 1.2 kg/min decreased individual milking duration and alleviated machine-milking-induced short-term changes to the teat tissue without adversely affecting milking performance or somatic cell count. Future studies are warranted to investigate the effect on milk production and udder health over a whole lactation period.

Evaluation of powdered 0.5% chlorhexidine acetate-based postmilking teat dip compared with a foamed 1% iodine-based postmilking teat dip under cold weather conditions in northern New York.

The objective of this trial was to compare a powdered 0.5% chlorhexidine acetate-based postmilking teat dip with a foamed 1% iodine-based postmilk teat dip during winter on clinical mastitis, subclinical mastitis (somatic cell count ≥200,000 cells/mL), linear score, teat skin condition, teat end score for hyperkeratosis, and risk of developing a new intramammary infection (IMI). Holstein cows (n = 331) housed in freestall and tiestall barns on one farm were blocked by pen, parity, lactation stage, and lactation performance. They were assigned randomly to a powdered chlorhexidine postmilking teat dip (PD; Derma Soft n' Dry, IBA Inc., Millbury, MA) or a foamed iodine-based postmilking teat dip (ID; FS-103, IBA Inc.). Treatments were applied for 6 wk starting January 4, 2016, for 3 milkings per day. Milk samples were collected from each quarter at the beginning and end of the trial and analyzed for aerobic culture and somatic cell count. Cows that had a clinical mastitis event during the trial were quarter sampled for aerobic culture at the time of clinical event. Teat skin condition and teat end score for hyperkeratosis were evaluated at the beginning, middle, and end of the trial based on a 3- and 5-point scale, respectively. No treatment difference was observed for linear score or teat skin condition. Teat end score was greater for ID cows compared with PD cows (2.72 vs. 2.77) at the conclusion of the trial. At the beginning of trial 102 PD quarters and 129 ID quarters had an IMI identified on aerobic culture, 402 PD and 457 ID quarters cultured negative, and 109 PD and 125 ID samples were classified as "no significant growth." At the conclusion of the trial, 129 PD and 101 ID quarters had an IMI. Use of PD resulted in a greater risk for developing a new IMI, based on bacteriological culture, at the conclusion of the trial as compared with ID (relative risk = 1.51; confidence interval: 1.10-2.07). Additionally, use of PD resulted in a greater risk as compared with ID of coagulase-negative staphylococci (relative risk = 1.5; confidence interval: 1.10-2.25) and Staphylococccus aureus (relative risk = 2.30; confidence interval: 1.04-5.07) to be present at the conclusion of the trial. In conclusion, use of PD led to a lower teat end score, an increase in new IMI, and an increased risk of coagulase-negative staphylococci and Staph. aureus compared with ID after 6 wk of product use.

Technical note: Evaluation of digital refractometers to estimate serum immunoglobulin G concentration and passive transfer in Jersey calves.

Previous data have demonstrated that refractometers can be used to estimate serum IgG, and that a cut-point of 7.8% Brix should be used to identify failure of passive transfer (FPT) in 1-d-old Holstein calves. The objective of the present study was to validate the use of refractometry to estimate serum IgG concentrations and evaluate FPT in Jersey calves. Blood samples (n = 97) were obtained from 1- to 3-d-old Jersey calves and centrifuged at 3,300 × g for 20 min at 25°C. Serum was analyzed for % Brix, total protein (TP), and refractive index (nD) using a Sper Scientific Digital Refractometer (model #300036, Sper Scientific, Scottsdale, AZ) within 12 h of sampling. Samples were then frozen and later analyzed in the laboratory for IgG by radial immunodiffusion. The mean serum IgG concentration for all calves was 23.7 mg/mL (SD = 12.5), with a range of 2.3 to 65.5 mg/mL. Mean serum % Brix was 8.9 (SD = 1.1; range 6.5 to 12.0). Serum % Brix was moderately correlated with IgG concentration (r = 0.77). Total protein and IgG were moderately correlated (r = 0.790). Regression was used to determine cut-points for approximately 10, 12, and 14 mg of IgG/mL and to determine the sensitivity and specificity of refractometry to identify FPT (serum IgG <10 mg/mL at 24 h of life). Brix cut-points analyzed were 7.1, 7.3, and 7.6%; TP cut-points were 4.6, 5.0, and 5.5 g/dL; and nD cut-points were 1.34332, 1.34271, and 1.3448, respectively, for 10, 12, and 14 mg of IgG/mL. The 7.3% Brix and 4.6 g/dL TP cut-points resulted in the greatest percentage of samples being correctly classified. These data suggest that digital refractometry is an acceptable and rapid method to estimate immunoglobulin G in Jersey calf serum.

Validating a refractometer to evaluate immunoglobulin G concentration in Jersey colostrum and the effect of multiple freeze-thaw cycles on evaluating colostrum quality.

The objectives of this study were to (1) validate a method using refractometry to rapidly and accurately determine immunoglobulin (IgG) concentration in Jersey colostrum, (2) determine whether there should be different refractive index (nD) and %Brix cut points for Jersey colostrum, and (3) evaluate the effect of multiple freeze-thaw (FT) cycles on radial immunodiffusion (RID) and a digital refractometer to determine IgG concentration in Jersey colostrum. Samples (n=58; 3L) of colostrum were collected from a dairy in northwestern Iowa. Samples were analyzed within 2h of collection for IgG concentration by RID, %Brix, and nD by refractometer and an estimate of IgG by colostrometer. Samples were frozen, placed on dry ice, and transported to the laboratory at Iowa State University (Ames). Samples arrived frozen and were placed in a -20°C manual-defrost freezer until further analysis. On d 7 (1FT), d 14 (2FT), and 1yr (3FT) all samples were thawed, analyzed for IgG by RID, %Brix, nD by refractometer, and IgG estimate by colostrometer, and frozen until reanalysis at the next time point. Fresh colostrum had a mean (±SD) IgG concentration of 72.91 (±33.53) mg/mL, 21.24% (±4.43) Brix, and nD 1.3669 (±0.0074). Multiple FT cycles did affect IgG as determined by RID and colostrometer reading. The IgG concentrations were greater in fresh and 1FT samples as compared with 2FT and 3FT samples (72.91, 75.38, 67.20, and 67.31mg of IgG/mL, respectively). The colostrometer reading was lower in 1FT samples compared with fresh and 2FT samples. Multiple FT cycles had no effect on nD or %Brix reading. In fresh samples, IgG concentration was moderately correlated with nD (r=0.79), %Brix (r=0.79), and colostrometer reading (r=0.79). Diagnostic test characteristics using the recommended cut point of 1.35966 nD resulted in similar sensitivities for 1FT and 2 FT samples (94.87 and 94.74%, respectively). Cut points of 18 and 19% Brix resulted in the greatest sensitivities (92.31 and 84.62%) and specificity (94.74 and 94.74%, respectively). The 18% Brix cut point resulted in 94.83% of the samples being correctly classified based on IgG concentration. These data support the use of digital refractometer to accurately and rapidly determine IgG concentration in fresh Jersey colostrum. Additionally, these data suggest that IgG concentration determined by RID is affected by multiple FT cycles, whereas estimates obtained by refractometer are not affected by multiple FT cycles.

Estimate of serum immunoglobulin G concentration using refractometry with or without caprylic acid fractionation.

Objectives of this study were to develop a rapid calf-side test to determine serum IgG concentrations using caprylic acid (CA) fractionation, followed by refractometry of the IgG-rich supernatant and compare the accuracy of this method with results obtained using refractometry using raw serum. Serum samples (n=200) were obtained from 1-d-old calves, frozen (-20°C), and shipped to the laboratory. Samples were allowed to thaw for 1h at room temperature. Fractionation with CA was conducted by adding 1mL of serum to a tube containing 45, 60, or 75µL of CA and 0.5, 1.0, or 1.5mL of 0.06 M acetic acid. The tube contents were mixed well, allowed to react for 1 min, and then centrifuged at 3,300 × g for 0, 10, or 20 min at 25°C. The %Brix and refractive index of the fractionated supernatant were determined using a digital refractometer. Nonfractionated serum was analyzed for %Brix (BRn), refractive index (nDn), and IgG concentration by radial immunodiffusion. The mean serum IgG concentration was 19.0 mg/mL [standard deviation (SD)=9.7], with a range of 3.5 to 47.0 mg/mL. The mean serum BRn was 8.6 (SD=0.91), with a range of 6.8 to 11.0. The mean serum nDn was 1.34566 (SD=0.00140), with a range of 1.34300 to 1.34930. Serum nDn was positively correlated with IgG concentration (correlation coefficient=0.86; n=185). Fractionated samples treated with 1mL 0.6 M acetic acid and 60µL of CA and not centrifuged before analysis resulted in a strong relationship between the refractive index of the fractionated supernatant and IgG (correlation coefficient=0.80; n=45). Regression was used to determine cut points indicative of 10, 12, and 14 mg of IgG/mL to determine the sensitivity and specificity of refractometry to identify failure of passive transfer (serum IgG <10 mg/mL at 24 h old). The nDn were 1.34414, 1.34448, and 1.34480 to predict 10, 12, and 14 mg of IgG/mL of serum, respectively. The BRn cut points were 7.6, 7.8, and 8.0, respectively. The nDn cut points of 1.34448 and 1.34480 resulted in similar specificities (82.9%), whereas the 1.34414 cut point had a specificity of 60.0%. The BRn cut point of 7.6 and 7.8%Brix resulted in a similar percentage of correctly classified samples (89.7 and 90.8%, respectively); however, the 7.8% Brix cut point resulted in fewer false positives. These results suggest that Brix refractometry of nonfractionated calf serum provides a strong estimate of IgG concentration and 7.8% Brix may be used as the cut point to identify failure of passive transfer in 1-d-old calves.

Estimate of colostral immunoglobulin G concentration using refractometry without or with caprylic acid fractionation.

Our objectives were to evaluate the use of refractometry as a means of estimating immunoglobulin G (IgG) concentration of bovine maternal colostrum (MC) and determine if fractionation of MC using caprylic acid (CA) improved estimates of IgG. Samples (n=85) of MC were collected from a single dairy in California and used to determine the method of CA fraction that produced the best prediction of IgG based on CA fractionation followed by refractometry. Subsequently, samples of MC (n=827) were collected from 67 farms in 12 states to compare refractometry with or without CA fractionation as methods to estimate IgG concentration. Samples were collected from the feeding pool and consisted of fresh (n=196), previously frozen (n=479), or refrigerated (n=152) MC. Samples were further classified by the number freeze-thaw cycles before analysis. Fractionation with CA was conducted by adding 1 mL of MC to a tube containing 75 µL of CA and 1 mL of 0.06 M acetic acid. The tube was shaken and allowed to react for 1 min. Refractive index of the IgG-rich supernatant (nDf) was determined using a digital refractometer. Whole, nonfractionated MC was analyzed for IgG by radial immunodiffusion (RID) and refractive index (nDw). The relationship between nDf and IgG (r=0.53; n=805) was weak, whereas that between nDw and IgG was stronger (r=0.73; n=823). Fresh samples analyzed by refractometry that subsequently went through 1 freeze-thaw cycle before RID analysis resulted in the strongest relationship between IgG and nDf or nDw (r=0.93 and 0.90, respectively). The MC samples collected fresh on the farm but frozen 2 or more times before analysis by refractometry or RID had low correlations between IgG and nDf and nDw (r=0.09 and 0.01). Samples refrigerated or frozen on the farm before analysis had weaker relationships between RID and nDf or nDw (r=0.38 to 0.80), regardless of the number of freeze-thaw cycles. Breed and lactation number did not affect the accuracy of either test. These results indicated that refractometry, without or with CA fractionation, was an accurate and rapid method to determine IgG concentration when samples of MC were not previously stored before refractometry and frozen only once before RID analysis.

Nationwide evaluation of quality and composition of colostrum on dairy farms in the United States.

The objective of this study was to characterize the quality of maternal colostrum (MC) fed to newborn dairy calves in the United States and identify the proportion of MC that meets industry standards for IgG concentration and total plate count (TPC). Samples of MC (n=827) were collected from 67 farms in 12 states between June and October 2010. Samples were collected from Holsteins (n=494), Jerseys (n=87), crossbred (n=7), and unidentified dairy cattle (n=239) from first (n=49), second (n=174), third or greater (n=128), and unknown (n=476) lactations. Samples were identified as fresh (n=196), refrigerated (n=152), or frozen (n=479) before collection, as well as whether the sample was from an individual cow (n=734) or pooled (n=93). Concentration of IgG in MC ranged from <1 to 200mg/mL, with a mean IgG concentration of 68.8 mg/mL (SD=32.8). Almost 30% of MC contained <50 mg of IgG/mL. The IgG concentration increased with parity (42.4, 68.6, and 95.9 mg/mL in first, second, and third and later lactations, respectively). No differences in IgG concentration were observed among breeds or storage method; however, IgG was highest in samples collected in the Midwest and lowest in samples collected in the Southwest (79.7 vs. 64.3 mg/mL). Total plate count of samples ranged from 3.0 to 6.8 log(10) cfu/mL, with a mean of 4.9 log(10) cfu/mL (SD=0.9) and was greater in samples collected in the Southeast compared with other regions of the country. Pooled samples had greater TPC than individual samples and refrigerated samples had greater TPC than frozen and fresh samples. Almost 43% of samples collected had TPC >100,000 cfu/mL, 16.9% of the samples had >1 million cfu/mL. Only 39.4% of the samples collected met industry recommendations for both IgG concentration and TPC. Almost 60% of MC on dairy farms is inadequate, and a large number of calves are at risk of failure of passive transfer or bacterial infections, or both. Also, the data indicate that regional differences exist in colostrum quality.

Anionic salts in the prepartum diet and addition of sodium bicarbonate to colostrum replacer, and their effects on immunoglobulin G absorption in the neonate.

The objectives of this experiment were to determine whether feeding anionic salts to prepartum Holstein cows affected their calf's colostral IgG passive transfer and whether adding sodium bicarbonate to a colostrum replacer (CR) would increase the efficiency of IgG absorption. Forty Holstein cows and their resulting calves were assigned to a 2 x 2 factorial arrangement of treatments in a randomized complete block design based on expected date of calving. Three weeks before the projected due date, cows were placed on 1 of 2 treatments: a diet without anionic salts (dietary cation-anion difference of +77 mEq/kg) or a diet with anionic salts (dietary cation-anion difference of -100 mEq/kg). Within 45 min after birth, all calves received 1 dose of a commercially available CR (132g of IgG) without or with supplemental sodium bicarbonate (19.5 g/dose). A half-dose of CR (66g of IgG) and sodium bicarbonate (9.75g) was fed at 6h of age. Calves received milk replacer at 12, 24, 36, and 48h. Blood samples were obtained from calves at 0, 6, 12, 24, and 48h and were analyzed for IgG concentration. Cows fed the diet supplemented with anionic salts had lower DMI on d 8, 5, 4, and 1 and lower urine pH 2 and 1 wk before parturition compared with cows fed the diet without supplemental anionic salts. Calves born from dams receiving anionic salts had similar IgG concentrations (15.1 vs. 14.4g/L) and apparent efficiency of absorption values (29.2 vs. 28.2%) compared with calves born from dams not fed anionic salts. Calves receiving supplemental sodium bicarbonate in the CR had higher serum IgG concentrations at 12 (14.4 vs. 12.0g/L), 24 (16.3 vs. 13.2g/L), and 48h (14.6 vs. 11.2g/L) and higher apparent efficiency of absorption values (31.2 vs. 26.1%) than calves that did not receive sodium bicarbonate in the CR. Calves receiving sodium bicarbonate also had greater area under the curve values for IgG absorption compared with calves not receiving sodium bicarbonate. There was a trend for an interaction with calves born from dams fed anionic salts having a greater area under the curve when fed supplemental sodium bicarbonate. Of the 40 calves in the study, 90% obtained adequate passive transfer (serum IgG > or = 10g/L). This study indicates that feeding anionic salts to the dam has no effect on passive transfer, whereas adding sodium bicarbonate to the CR increased IgG uptake in calves.