Effect of feeding heat-treated colostrum on risk for infection with Mycobacterium avium ssp. paratuberculosis, milk production, and longevity in Holstein dairy cows.

In summer 2007, a randomized controlled field trial was initiated on 6 large Midwest commercial dairy farms to investigate the effect of feeding heat-treated (HT) colostrum on transmission of Mycobacterium avium ssp. paratuberculosis (MAP) and on future milk production and longevity within the herd. On each farm, colostrum was collected daily from fresh cows, pooled, divided into 2 aliquots, and then 1 aliquot was heat-treated in a commercial batch pasteurizer at 60°C for 60min. A sample from each batch of colostrum was collected for PCR testing (MAP-positive vs. MAP-negative). Newborn heifer calves were removed from the dam within 30 to 60min of birth and systematically assigned to be fed 3.8 L of either fresh (FR; n=434) or heat-treated (HT; n=490) colostrum within 2h of birth. After reaching adulthood (>2 yr old), study animals were tested once annually for 3 yr (2010, 2011, 2012) for infection with MAP using serum ELISA and fecal culture. Lactation records describing milk production data and death or culling events were collected during the 3-yr testing period. Multivariable model logistic and linear regression was used to investigate the effect of feeding HT colostrum on risk for testing positive to MAP during the 3-yr testing period (positive/negative; logistic regression) and on first and second lactation milk yield (kg/cow; linear regression), respectively. Cox proportional hazards regression was used to investigate the effect of feeding HT colostrum on risk and time to removal from the herd. Fifteen percent of all study animals were fed PCR-positive colostrum. By the end of the 3-yr testing period, no difference was noted in the proportion of animals testing positive for MAP, with either serum ELISA or fecal culture, when comparing the HT group (10.5%) versus the FR group (8.1%). There was no effect of treatment on first- (HT=11.797kg; FR=11,671kg) or second-lactation (HT=11,013kg; FR=11,235kg) milk production. The proportion of cows leaving the herd by study conclusion was not different for animals originally fed HT (68.0%) versus FR (71.7%) colostrum. Although a previous study showed that feeding HT colostrum (60°C for 60min) produces short-term benefits, including improved passive transfer of IgG and reduced morbidity in the preweaning period, the current study found no benefit of feeding HT colostrum on long-term outcomes including risk for transmission of Mycobacterium avium ssp. paratuberculosis, milk production in the first and second lactation, and longevity within the herd.

Effects of weekly regrouping of prepartum dairy cows on metabolic, health, reproductive, and productive parameters.

The objectives of the current experiment were to determine the effect of 2 prepartum grouping strategies on the health, metabolic, reproductive, and productive parameters of dairy cows. Jersey cows enrolled in the experiment at 253±3 d of gestation (d 0=calving) were balanced for parity and projected 305-d mature equivalent and assigned to 1 of 2 treatments. Cows assigned to the traditional (TRD; n=6 replicates with a total of 308 cows) treatment were moved to the study pen as a group of 44 cows and weekly thereafter groups of 2 to 15 cows were moved to the study pen to reestablish stocking density. Cows assigned to the all-in-all-out (AIAO; n=6 replicates with a total of 259 cows) treatment were moved to the study pen in groups of 44 cows, but no new cows entered the AIAO pen until the end of the replicate. At the end of each replicate, a new TRD and AIAO group started but pens were switched. Cows were milked thrice daily and monthly milk yield, fat and protein contents, and somatic cell count data were recorded up to 305 d postpartum. Plasma nonesterified fatty acid concentration was measured weekly from d -18±3 to 24±3 and plasma ß-hydroxybutyrate was measured weekly from d 3±3 to 24±3. Cows were examined on d 1, 4±1, 7±1, 10±1, and 13±1 for diagnosis of uterine diseases and had their ovaries scanned by ultrasound on d 39±3 and 53±3 to determine resumption of ovarian cycles. Average stocking density was reduced for the AIAO (71.9%) treatment compared with the TRD (86.9%) treatment. Treatment did not affect the incidences of retained fetal membranes (TRD=10.9, AIAO=11.6%), metritis (TRD=16.7, AIAO=19.8%), and acute metritis (TRD=1.7, AIAO=3.6%). Concentrations of nonesterified fatty acids (TRD=80.4±8.2, AIAO=62.9±8.5 µmol/L) and ß-hydroxybutyrate (TRD=454.4±10.9, AIAO=446.1±11.1 µmol/L) were not different between treatments. Percentages of cows that resumed ovarian cycles by d 39±3 (TRD=70.8, AIAO=63.1%) and 53±3 (TRD=90.1, AIAO=90.2%) were not different between treatments. Similarly, treatment had no effect on rate of removal from the herd {TRD=referent, AIAO [(adjusted hazard ratio (95% confidence interval)]=0.85 (0.63, 1.15)} or rate of pregnancy [TRD=referent, AIAO=1.07 (0.88, 1.30)]. Finally, treatment did not affect energy-corrected milk yield (TRD=34.4±0.6, AIAO=34.3±0.7 kg/d). In conditions of adequate feed bunk space, the AIAO treatment did not improve health, metabolic, reproductive, or productive parameters compared with the TRD treatment.

Heat-treated colostrum and reduced morbidity in preweaned dairy calves: results of a randomized trial and examination of mechanisms of effectiveness.

A randomized controlled clinical trial was conducted using 1,071 newborn calves from 6 commercial dairy farms in Minnesota and Wisconsin, with the primary objective being to describe the effects of feeding heat-treated colostrum on serum immunoglobulin G concentration and health in the preweaning period. A secondary objective was to complete a path analysis to identify intermediate factors that may explain how feeding heat-treated colostrum reduced the risk for illness. On each farm, colostrum was collected each day, pooled, and divided into 2 aliquots; then, one aliquot was heat-treated in a commercial batch pasteurizer at 60°C for 60 min. Samples of fresh and heat-treated colostrum were collected for standard microbial culture (total plate count and total coliform count, cfu/mL) and for measurement of immunoglobulin G concentrations (mg/mL). Newborn calves were removed from the dam, generally within 30 to 60 min of birth, and systematically assigned to be fed 3.8L of either fresh (FR, n=518) or heat-treated colostrum (HT, n=553) within 2h of birth. Venous blood samples were collected from calves between 1 and 7d of age for measurement of serum IgG concentrations (mg/mL). All treatment and mortality events were recorded by farm staff between birth and weaning. Regression models found that serum IgG concentrations were significantly higher in calves fed HT colostrum (18.0 ± 1.5 mg/mL) compared with calves fed FR colostrum (15.4 ± 1.5 mg/ml). Survival analysis using Cox proportional hazards regression indicated a significant increase in risk for a treatment event (any cause) in calves fed FR colostrum (36.5%, hazard ratio=1.25) compared with calves fed HT colostrum (30.9%). In addition, we observed a significant increase in risk for treatment for scours in calves fed FR colostrum (20.7%, hazard ratio=1.32) compared with calves fed HT colostrum (16.5%). Path analysis suggested that calves fed HT colostrum were at lower risk for illness because the heat-treatment process caused a significant reduction in colostrum total coliform count, which was associated with a reduced risk for illness as a function of improved serum IgG concentrations.

Heat treatment of colostrum on commercial dairy farms decreases colostrum microbial counts while maintaining colostrum immunoglobulin G concentrations.

This study was conducted on 6 commercial dairy farms in Minnesota and Wisconsin to describe the effect of heat treatment (at 60°C for 60 min) on colostrum, on colostrum bacteria counts, and immunoglobulin G concentrations. First-milking colostrum was collected each day, pooled, divided into 2 aliquots, and then 1 aliquot was heat treated in a commercial batch pasteurizer at 60°C for 60 min. Frozen samples of pre- and post- heat-treated colostrum were submitted for standard microbial culture (total plate count and total coliform count, cfu/mL) and testing for immunoglobulin G concentrations (mg/mL). Data were analyzed from 266 unique batches of colostrum. Linear regression showed that heat treatment decreased colostrum total plate counts (-2.25 log(10)) and coliform counts (-2.49 log(10)), but, overall, did not affect colostrum IgG concentration. Though higher-quality batches of colostrum did experience a greater magnitude of loss of IgG as a result of heat treatment as compared with lower- or intermediate-quality batches of colostrum, the colostral IgG concentrations in these batches remained high overall, and within acceptable limits for feeding. This study demonstrates that batch heat treatment of colostrum at 60°C for 60 min can be successfully conducted on commercial dairy farms by farm staff to decrease colostrum microbial counts while maintaining colostrum IgG concentrations.

Animal welfare in cross-ventilated, compost-bedded pack, and naturally ventilated dairy barns in the upper Midwest.

The objective of this cohort study was to investigate animal welfare in 2 newer dairy housing options in the upper Midwest, cross-ventilated freestall barns (CV) and compost-bedded-pack barns (CB), compared with conventional, naturally ventilated freestall barns (NV). The study was conducted on 18 commercial dairy farms, 6 of each housing type, in Minnesota and eastern South Dakota. The primary breed in all farms was Holstein; 1 CV and 1 NV herd had approximately 30% Jersey-Holstein crossbreds. All freestall herds used sand for bedding. Farms were visited 4 times (once in each season) between January and November 2008, and approximately 93% of all animals in each pen were visually scored on each visit. Outcome-based measurements of welfare (locomotion, hock lesions, body condition score, hygiene, respiration rates, mortality, and mastitis prevalence) were collected on each farm. Lameness prevalence (proportion of cows with locomotion score ≥3 on a 1 to 5 scale, where 1=normal and 5=severely lame) in CB barns (4.4%) was lower than that in NV (15.9%) and CV (13.1%) barns. Lameness prevalence was similar between CV and NV barns. Hock lesion prevalence (proportion of cows with a lesion score ≥2 on a 1 to 3 scale, where 1=normal, 2=hair loss, and 3=swelling) was lower in CB barns (3.8%) than in CV (31.2%) and NV barns (23.9%). Hygiene scores (1 to 5 scale, where 1=clean and 5=very dirty) were higher for CB (3.18) than CV (2.83) and NV (2.77) barns, with no differences between CV and NV barns. Body condition scores, respiration rates, mastitis prevalence, culling, and mortality rates did not differ among housing systems. The CV and NV barns were evaluated using the cow comfort index (proportion of cows lying down in a stall divided by all animals touching a stall) and the stall usage index (proportion of cows lying divided by all animals in the pen not eating). The CV barns tended to have greater cow comfort index (85.9%) than the NV barns (81.4%) and had greater stall usage index (76.8% and 71.5%, respectively). Dairy cattle housed in CB barns had reduced lameness and hock lesions compared with those housed in freestall barns and had no adverse associations with body condition, respiration rates, mastitis prevalence, culling, or mortality. When comparing the 2 freestall housing options, CV barns had improved cow comfort indices compared with NV barns. Although cows in CB barns had better feet and leg health, as indicated by the reduced lameness and hock lesion prevalence, acquiring bedding and managing the bedded pack could limit their use.

Sex-sorted semen for dairy heifers: effects on reproductive and lactational performances.

The objectives of this study were to evaluate the effects of using sex-sorted semen for first AI of heifers on health and productivity during first lactation. Holstein heifers (herd A=227 and herd B=1,144) received first artificial insemination (AI) with sex-sorted semen (SX; n=343) or conventional semen (CS; n=1,028), and all heifers that displayed estrus after first AI were reinseminated with conventional semen up to 11 times before being culled. Age at first AI was 13.1+/-0.1 and 13.8+/-0.1 mo for SX and CS heifers, respectively, in herd A and 12.9+/-0.1 mo for both SX and CS heifers in herd B. Pregnancy per AI after first AI was greater for CS heifers than for SX heifers (51.8 vs. 40.2%). From heifers initially enrolled, 70.2% calved in herds A (n=188) or B (n=774) and first-lactation data were collected. Interval from first AI to calving was greater for SX heifers than for CS heifers (10.2+/-0.1 vs. 9.9+/-0.1 mo). Among heifers conceiving to first AI, SX heifers were more likely than CS heifers to deliver a female calf (85.7 vs. 47.7%), but because SX heifers were more likely to deliver a dead calf (8.8 vs. 3.4%), the difference in proportion of SX and CS heifers delivering a live female calf was smaller than expected (SX=79.1%; CS=47.2%). Rearing cost from first AI to calving was greater for SX heifers than for CS heifers ($775.3+/-6.7 vs. $750.0+/-5.9), but calf revenue tended to be greater for SX heifers ($142.0+/-7.2 vs. $126.7+/-6.4) and cost per female calf produced was smaller for SX heifers than for CS heifers ($-809.4+/-10.8 vs. $-1,249.7+/-10.9). Treatment did not affect calving difficulty, proportion of heifers needing assistance, and incidence of retained fetal membranes or metritis. Among heifers that conceived to first AI, however, SX heifers were more likely to be culled within 30 DIM (3.3 vs. 1.6%) and tended to be more likely to be culled within 60 DIM (5.5 vs. 3.4%) than CS heifers, but overall replacement cost was not different ($136.8+/-13.4). Total milk yield (9,245.5+/-84.7 kg) and income over feed cost ($554.7+/-5.1) were not different. Overall economic return was greater for SX heifers than CS heifers ($-83.7+/-36.7 vs. -175.3+/-33.4). Use of sex-sorted semen for first insemination of virgin heifers reduced the cost per female calf produced and increased the economic return during the first lactation.

Loss of income from cows shedding Mycobacterium avium subspecies paratuberculosis prior to calving compared with cows not shedding the organism on two Minnesota dairy farms.

Quantification of the financial effect of Mycobacterium paratuberculosis infection on lactation performance is essential to encourage participation of dairy cattle producers in Johne's disease (JD) control programs. The objective of this study was to evaluate the differences in net income per lactation of cows shedding Mycobacterium paratuberculosis before calving compared with test-negative cows. Two Minnesota dairies were enrolled in the study and fecal samples were collected from 1,048 cows during the close-up period. Milk production, clinical diseases (other than clinical JD), and reproductive performance data were recorded for each cow. Overall, fecal-culture-positive (FCP) cows produced 1,355 kg less than fecal-culture-negative (FCN) cows. Fecal-culture-positive cows that survived their current lactation produced $276 less in milk income than cows that were FCN ($1,956 vs. $1,680; SD $526, $570). Fecal-culture-positive cows were 3.0 (95% confidence interval: 1.6-5.8) times more likely to be culled than FCN cows. The mean days open (number of days from calving to conception) was not statistically significant and the cost differences for clinical disease other than JD were small and neither statistically nor economically significant between FCP and FCN cows. Among all FCP cows, income over feed costs losses were $366 per cow per lactation compared with FCN cows. Among FCP nonculled cows, income over feed costs losses were $276 more compared with FCN cows and this difference was statistically significant. There was a total loss of $155 per lactation for nonculled FCP cows retained in the herd compared with FCN cows retained in the herd. Among culled cows, FCP cow losses were $50 less because of age at culling and $120 for reduced beef value. This totaled a loss of $441 for culled FCP cows compared with culled FCN cows. The losses as a result of lower lactation performance and early culling from the herd should alarm dairy producers and motivate them to implement the appropriate control measures for the disease.

Exploring the impact of sexed semen on the structure of the dairy industry.

Widespread commercial application of sexed semen is expected within the next decade because of continued improvements in fertility of sexed semen and sorting capacity. The objective of this study was to explore the potential impact of widespread application of sexed semen on the structure of the dairy industry in the United States. Historically, female offspring from all heifers and cows were needed to produce enough dairy replacement heifers to replace culled cows. The use of sexed semen allows for a decoupling of breeding decisions necessary to obtain an adequate supply of dairy replacement heifers from those needed to achieve pregnancies needed to start new lactations. Application of sexed semen allows dairy producers to select among their herds' potential dams and produce dairy replacement heifers from only the genetically superior animals. The rate of genetic progress is expected to increase, but not more than 15% of the rate of gain accomplished through sire selection achieved through conventional (nonsexed) artificial insemination breeding. The supply of dairy replacement heifers is expected to grow to meet and temporarily exceed current demand, resulting in reduced prices for dairy replacement heifers. Consequently, herd turnover rates are expected to increase slightly, and herd expansions may accelerate. The rate of consolidation of dairy farms is expected to increase. Widespread application of sexed semen may temporarily increase the supply of milk, which would result in lower milk prices. The cost of milk production will be reduced as well. Many breeding options exist for the genetically poorer cows in the herd. The optimal breeding mix depends on the value of the various kinds of calves that could be produced. More crossbred calves for beef production may be produced; however, a market for these crossbred calves is not well established. Increased specialization is expected with more dairy producers deciding not to raise their own heifers but to purchase replacements. Other dairy farms might specialize in producing genetically superior dairy replacement heifers for sale. Depending on the value of calves not raised for replacements, artificial insemination organizations might market beef conventional semen or beef male sexed semen to dairy farms. The use of sexed semen should lower the cost of progeny-testing programs and embryo transfer and enhance the value of genetic markers. Eventually, the economic benefits from the use of sexed semen will be passed on to consumers.

Evaluation of the Dairy Comp 305 module "Cow Value" in two Ontario dairy herds.

The study was conducted to evaluate how the "Cow Value" module of Dairy Comp 305 (Valley Agricultural Software, Tulare, CA) performed under commercial conditions. The "Cow Value" module, COWVAL, computes a farm-specific net present value relative to an average replacement heifer for each cow in the milking and dry herd, which allows a ranking of the cows on the farm compared with replacing her with a typical replacement heifer on that farm. The average replacement heifer is used as the baseline for comparison and has a COWVAL of $0. Retaining a cow with a negative COWVAL is projected to be less profitable than replacing that cow with a new heifer. The objectives of the study were to explore trends in COWVAL over and during multiple lactations for the same cows; to describe factors that influence changes in COWVAL from one monthly Dairy Herd Improvement test to the next; and to evaluate the behavior of COWVAL after it drops below a baseline of $0 during the lifetime of a cow. Monthly Dairy Comp 305 backup cow files from 2 On-tario dairy herds between December 1999 and Decem-ber 2005 were used to generate COWVAL and list production, reproduction, and disease data for the milking cows. In total, 1,463 cows and 20,071 tests were analyzed. Within the first 60 d in milk (DIM), COWVAL was unstable and showed large fluctuations over a range of several thousand Canadian dollars (Can$). After 60 DIM COWVAL was relatively stable. The variability from month to month became less as the lactation progressed and the risk of a change in reproductive status decreased. The reproductive status of the cow influ-enced COWVAL: fresh, open, and pregnant cows had a greater COWVAL than cows declared "do not breed." As parity increased, there was a tendency toward lower COWVAL and smaller monthly changes in COWVAL. The COWVAL of 170 cows dropped below the baseline of $0 after 60 DIM. The COWVAL of 54% of those cows remained below $0, whereas 31.6% had a subsequent COWVAL > $500 (Can$). Farm management should not rely exclusively on COWVAL for culling decisions, particularly for cows that have not had at least 3 milk tests.

Passive transfer of immunoglobulin G and preweaning health in Holstein calves fed a commercial colostrum replacer.

The objective of this study was to describe passive transfer of IgG and preweaning health in newborn calves fed a commercially available plasma-derived colostrum replacement (CR) product or maternal colostrum (MC). Twelve commercial Holstein dairy farms enrolled singleton newborn heifer calves to be fed fresh MC (n = 239 calves) or one dose of CR containing 125 g of Ig (n = 218 calves) as the first colostrum feeding. For 7 of these farms that routinely provided a second feeding of 1.9 L of MC to their calves 8 to 12 h after the first colostrum feeding, calves assigned to the CR treatment group were offered a second feeding consisting of 1.9 L of commercial milk replacer supplemented with one dose of a commercially available plasma-derived colostrum supplement, containing 45 g of Ig per dose, 8 to 12 h after the first colostrum feeding. A blood sample was collected from all calves between 1 to 8 d of age for serum IgG and total protein (TP) determination, and records of all treatment and mortality events were collected until weaning. Serum IgG and TP concentrations were significantly higher in calves fed MC (IgG = 14.8 +/- 7.0 mg/mL; TP = 5.5 +/- 0.7 g/dL) compared with calves fed CR (IgG = 5.8 +/- 3.2 mg/mL; TP = 4.6 +/- 0.5 g/dL). The proportion of calves with failure of passive transfer (serum IgG <10.0 mg/mL) was 28.0 and 93.1% in the MC and CR treatment groups, respectively. Though a trend was present, the proportion of calves treated for illness was not statistically different for calves fed MC (51.9%) vs. CR (59.6%). Total number of days treated per calf (MC = 1.7; CR = 2.0), treatment costs per calf (MC = $10.84; CR = $11.88), and proportion of calves dying (MC = 10.0%; CR = 12.4%) was not different between the 2 colostrum treatment groups. The mean serum total protein concentration predictive of successful passive transfer (serum IgG = 10 mg/mL) was 5.0 g/dL in calves fed MC or CR. Long-term follow-up of these calves (to maturity) is ongoing to describe the effects of feeding CR on longevity, productivity, risk for Johne's disease, and economics.

Heat-treatment of bovine colostrum. II: effects of heating duration on pathogen viability and immunoglobulin G.

Batches (30-L) of first-milking bovine colostrum, inoculated with Mycoplasma bovis (10(8) cfu/mL), Listeria monocytogenes (10(6) cfu/mL), Escherichia coli O157:H7 (10(6) cfu/mL), Salmonella enteritidis (10(6) cfu/mL), and Mycobacterium avium subsp. paratuberculosis (Map; 10(3) cfu/mL), were heat-treated at 60 degrees C for 120 min in a commercial on-farm batch pasteurizer system. Duplicate 50-mL subsamples of colostrum were collected at 15-min intervals throughout the heat-treatment process for the purpose of bacterial culture and for measurement of IgG concentration (mg/mL) and antibody activity [log2(bovine viral diarrhea virus type 1 serum neutralization titer)]. Four replicate batches of colostrum were run for each of the 5 pathogens studied. There was no effect of heating moderate- to high-quality colostrum at 60 degrees C for at least 120 min on mean IgG concentration (pre = 60.5 mg/mL; post = 59.1 mg/mL). Similarly, there was no effect of heat-treatment on the mean log2 bovine viral diarrhea virus type 1 serum neutralization titer (pre = 12.3; post = 12.0). Viable M. bovis, L. monocytogenes, E. coli O157:H7, and S. enteritidis added to colostrum could not be detected after the colostrum was heat-treated at 60 degrees C for 30 min. Average bacteria counts showed that Map was not detected when batches were heated at 60 degrees C for 60 min. Although the authors believe that heat-treating colostrum at 60 degrees C for 60 min should be sufficient to eliminate Map from colostrum in most situations, further research is needed to determine whether these findings may be replicated, given that variability was observed in Map culture results.

Invited review: Culling: nomenclature, definitions, and recommendations.

Replacing cows on a dairy is a major cost of operation. There is a need for the industry to adopt a more standardized approach to reporting the rate at which cows exit from the dairy, and to reporting the reasons why cows are replaced and their destination as they exit the dairy. Herd turnover rate is recommended as the preferred term for characterizing the cows exiting a dairy, in preference to herd replacement rate, culling rate, or percent exiting, all of which have served as synonyms. Herd turnover rate should be calculated as the number of cows that exit in a defined period divided by the animal time at risk for the population being characterized. The terms voluntary and involuntary culling suffer from problems of definition and their use should be discouraged. Destination should be recorded for all cows that exit the dairy and opportunities to record one or more reasons for exiting should be provided by management systems. Comparing reported reasons between dairies requires considerable caution because of differences in case definitions and recording methods. Relying upon culling records to monitor disease has been and will always be an ineffective management strategy. Dairies are encouraged to record and monitor disease events and reproductive performance and use this information as the basis for management efforts aimed at reducing the need to replace cows.

Heat treatment of bovine colostrum. I: effects of temperature on viscosity and immunoglobulin G level.

The objective of this study was to identify the critical temperature, at or below which heat-treatment of bovine colostrum would produce no significant changes in viscosity, IgG concentration, or Ig activity. Results of preliminary work, using a Rapid Visco Analyzer (RVA) to heat 50-mL aliquots from 6 unique batches of bovine colostrum at 59, 60, 61, 62, and 63 degrees C, suggested that colostrum could be heated to 60 degrees C for up to 120 min without changing viscosity or IgG concentration. This finding was confirmed by heating 50-mL aliquots from 30 unique batches of colostrum in an RVA for 120 min at 60 and 63 degrees C. Heating colostrum to 63 degrees C resulted in an estimated 34% decrease in IgG concentration and 33% increase in viscosity. However, there was no difference in IgG concentration between preheat-treated (73.4 +/- 26.5 mg/mL) and post-heat-treated (74.5 +/- 24.3 mg/mL) samples after heating colostrum to 60 degrees C in an RVA for 120 min. Similarly, viscosity was unaffected after heating colostrum to 60 degrees C in an RVA for 120 min. High quality colostrum (> or =73.0 mg/mL) suffered greater losses of IgG and greater viscosity changes when heated to 63 degrees C than did moderate quality colostrum (<73.0 mg/mL). However, the effects of colostrum quality were minor if high quality colostrum was only heated to 60 degrees C. The results of a bovine viral diarrhea serum neutralization assay suggested that antibody activity was unchanged after heating colostrum to either 60 or 63 degrees C. However, these results were interpreted as being inconclusive due to a high proportion of missing results because of the congealing of many samples after heat treatment. The results of this study indicate that 50-mL volumes of bovine colostrum can be heat treated at 60 degrees C for up to 120 min in an RVA without affecting IgG concentration or viscosity.

Preventing bacterial contamination and proliferation during the harvest, storage, and feeding of fresh bovine colostrum.

The objectives of this study were to identify control points for bacterial contamination of bovine colostrum during the harvesting and feeding processes, and to describe the effects of refrigeration and use of potassium sorbate preservative on bacteria counts in stored fresh colostrum. For objective 1, first-milking colostrum samples were collected aseptically directly from the mammary glands of 39 cows, from the milking bucket, and from the esophageal feeder tube. For objective 2, 15-mL aliquots of colostrum were collected from the milking bucket and allocated to 1 of 4 treatment groups: 1) refrigeration, 2) ambient temperature, 3) refrigeration with potassium sorbate preservative, and 4) ambient temperature with potassium sorbate preservative. Subsamples from each treatment group were collected after 24, 48, and 96 h of storage. All samples underwent bacteriological culture for total plate count and coliform count. Bacteria counts were generally low or zero in colostrum collected directly from the gland [mean (SD) log10 cfu/mL(udder) = 1.44 (1.45)]. However, significant bacterial contamination occurred during the harvest process [mean (SD) log10 cfu/mL(bucket) = 4.99 (1.95)]. No additional bacterial contamination occurred between the bucket and the esophageal feeder tube. Storing colostrum at warm ambient temperatures resulted in the most rapid increase in bacteria counts, followed by intermediate rates of growth in nonpreserved refrigerated samples or preserved samples stored at ambient temperature. The most effective treatment studied was the use of potassium sorbate preservative in refrigerated samples, for which total plate count and total coliform counts dropped significantly and then remained constant during the 96-h storage period.

Evaluation and use of three cowside tests for detection of subclinical ketosis in early postpartum cows.

The objective was to evaluate the performance of 3 cowside diagnostic tests for detection of subclinical ketosis, defined as a serum beta-hydroxybutyrate (BHBA) concentration >or=1400 micromol/L. On 16 d over a 5-mo period, samples of serum, milk, and urine were collected on a large dairy facility from cows of all parities between 2 and 15 DIM. The sample proportion of subclinical ketosis was 7.6% (n = 859 samples from 545 cows). The KetoCheck powder (Great States Animal Health, St. Joseph, MO) detecting acetoacetate in milk samples was very specific (99%) but poorly sensitive (41%). Respective sensitivities and specificities of the Ketostix strip detecting acetoacetate in urine samples (Bayer Corporation, Elkhart, IN) were 78 and 96% with a cut-off point of "small", or 49 and 99% with a cut-off of "moderate." The KetoTest strip (Sanwa Kagaku Kenkyusho Co. Ltd., Nagoya, Japan) using milk samples had a sensitivity and specificity of 73 and 96% with a cut-off of 100 micromol of BHBA/L or 27 and 99% with a cut-off of 200 micromol of BHBA/L. On average, use of the Ketostix at the "small" cut-off point or the KetoTest at 100 micromol/L would result in no more than 3 or 4 false positives per 100 cows screened, with prevalence levels ranging from 5 to 30%, whereas the number of false negatives would range from one false negative at 5% prevalence to 7 or 8 false negatives at 30% prevalence. Either the Ketostix or KetoTest strips would provide acceptable results for screening individual cows on commercial dairies to detect subclinical ketosis. Over this prevalence range, the KetoCheck powder test would have limited application as a screening test. Despite only one false positive per 100 animals screened, false negatives resulting from screening with the KetoCheck test would be too frequent, ranging from 3 false negatives at 5% prevalence to 18 at 30% prevalence in a population of 100 tested cows. Finally, given their relative imprecision, use of any of these individual cowside tests to estimate herd prevalence must be done cautiously, especially when only a small number of animals are sampled.

Effect of on-farm commercial batch pasteurization of colostrum on colostrum and serum immunoglobulin concentrations in dairy calves.

The objectives were to describe the effect of on-farm commercial batch pasteurization on immunoglobulin (IgG) concentrations and the fluid and feeding characteristics of colostrum and to compare serum IgG concentrations in calves fed fresh versus pasteurized colostrum. Newborn calves (123) were systematically allocated to dietary treatments of either fresh or pasteurized colostrum at both the first and second colostrum feedings. The IgG concentrations were measured for batches of colostrum fed fresh and in pre and postpasteurized samples for batches of colostrum fed after being pasteurized and in calf serum. Pasteurization reduced colostrum IgG concentration, with the percentage reduction averaging 58.5 and 23.6% for 95-L and 57-L batches, respectively. Pasteurizing high quality colostrum in 57-L (vs. 95-L) batches resulted in higher IgG concentrations in the end product. Pasteurization of 57-L batches produced colostrum of normal or only mildly thickened consistency that could be fed to calves. Serum IgG concentrations were higher for calves fed fresh colostrum and for calves with a shorter time interval (< or = 6 h) between first and second colostrum feedings. After controlling for the time interval between feedings, serum IgG concentrations were significantly higher for 40 calves fed unpasteurized (19.1 mg/ml) vs. 55 calves fed pasteurized colostrum (9.7 mg/ml) for calves fed 2 L at first feeding. By contrast, there was no difference in serum IgG concentrations between 8 calves fed unpasteurized (16.1 mg/ml) and 20 calves fed pasteurized colostrum (13.5 mg/ml) after calves were fed 4 L at the first feeding. While the latter results suggest that pasteurizing colostrum may work for producers with excellent colostrum management, these results are preliminary and should be interpreted with caution, given the fewer number of calves and batches of colostrum involved with this second comparison.

Effectiveness of an internal teat seal in the prevention of new intramammary infections during the dry and early-lactation periods in dairy cows when used with a dry cow intramammary antibiotic.

The objectives of this study were to determine the effect of infusion with an internal teat seal at dry off, when used as an adjunct to long-acting antibiotic infusion at dry off, on the risk for acquiring a new intramammary infection (IMI) during the dry period, prevalence of IMI and linear score (LS) after calving, and risk for experiencing a clinical mastitis event between dry off and 60 DIM. A total of 437 cows from 2 dairy herds, with no clinical mastitis and 4 functional quarters, were enrolled at dry off. Prior to the final milking, all quarters were sampled for bacteriological culture and SCC analysis. After milking, all 4 quarters were infused with a commercially available long-acting dry cow antibiotic. Two contralateral quarters were then infused with an internal teat seal (Orbeseal, Pfizer Animal Health, New York). Following calving the teat seal was stripped out at first milking. Duplicate milk samples were collected between 1 to 3 DIM and again between 6 to 8 DIM for culture and SCC analysis. Quarters treated with Orbeseal had significantly lower prevalence of IMI at 1 to 3 DIM (tx = 22.8%, control = 29.1%), had significantly fewer quarters that acquired a new IMI between dry off and 1 to 3 DIM (tx = 20.2%, control = 25.4%), and had significantly fewer quarters affected by a clinical mastitis event between dry off and 60 DIM (tx = 5.9%, control = 8.0%). Multivariable analysis showed a significant effect of treatment, with treated quarters being 30% less likely to develop a new IMI between dry off and 1 to 3 DIM, 31% less likely to have an IMI present at 1 to 3 DIM, 33% less likely to experience a clinical mastitis event between dry off and 60 DIM, and having significantly lower linear score measures at 1 to 3 DIM and 6 to 8 DIM, compared with control quarters.

The protein folding problem: a biophysical enigma.

Protein folding, the problem of how an amino acid sequence folds into a unique three-dimensional shape, has been a long-standing problem in biology. The success of genome-wide sequencing efforts has increased the interest in understanding the protein folding enigma, because realizing the value of the genomic sequences rests on the accuracy with which the encoded gene products are understood. Although a complete understanding of the kinetics and thermodynamics of protein folding has remained elusive, there has been considerable progress in techniques to predict protein structure from amino acid sequences. The prediction techniques fall into three general classes: comparative modeling, threading and ab initio folding. The current state of research in each of these three areas is reviewed here in detail. Efforts to apply each method to proteome-wide analysis are reviewed, and some of the key technical hurdles that remain are presented. Protein folding technologies, while not yet providing a full understanding of the protein folding process, have clearly progressed to the point of being useful in enabling structure-based annotation of genomic sequences.

Hatchery and transportation factors associated with early poult mortality in commercial turkey flocks.

A prospective ecologic study, using routinely collected data from commercial turkey companies, was done of 312 turkey flocks placed over a 1-yr period. The objective was to identify hatchery- and transportation-associated risk factors for poult mortality in the first 14 d after placement using statistical models. Logistic regression was used to determine the effects of hatchery-and transportation-related factors on odds of mortality for hen and tom flocks separately. Hatchery and transportation-related risk factors for hen flock mortality included truck, truck temperature, and number of poults dead on arrival at the farm. Hatchery and transportation-related risk factors for tom flock mortality included desnooding, truck, truck temperature, shipping time, and weather conditions at placement.

EPR-detected folding kinetics of externally located cysteine-directed spin-labeled mutants of iso-1-cytochrome c.

We report the application of our newly developed dielectric resonator-based flow and stopped-flow kinetic EPR systematically to probe protein folding in yeast iso-1-cytochrome c at cysteine-directed spin-labeled locations. The locations studied have not been previously directly probed by other techniques, and we observe them on a time scale stretching from 50 micros to seconds. On the basis of crystal structure and homology information, the following mutation-tolerant, externally located cysteine labeling sites were chosen (in helices, T8C, E66C, and N92C; in loops, E21C, V28C, H39C, D50C, and K79C), and labeling at these sites was not destabilizing. Dilution of denaturant was used to induce folding and thereby to cause a change in the spin label EPR signal as folding altered the motion of the spin label. Under folding conditions, including the presence of imidazole to eliminate kinetic trapping due to heme misligation, a phase of folding on the 20-30 ms time scale was found. This phase occurred not only at the T8C and N92C labeling sites in the N- and C-terminal helices, where such a phase has been associated with folding in these helices, but overall at labeling sites throughout the protein. In the absence of imidazole the 20-30 ms phase disappeared, and another phase having the time scale of 1 s appeared throughout the protein. There was evidence under all conditions for a burst phase on a scale of less than several milliseconds which occurred at labeling positions V28C, H39C, D50C, E66C, and K79C in the middle of the protein sequence. At spin-labeled D50C rapid-mix flow EPR indicated a very short approximately 50 micros phase possibly associated with the prefolding or compaction of the loop to which D50 belongs. Spin labels have been criticized as perturbing the phenomena which they measure, but our spin labeling strategy has reported common kinetic themes and not perturbed, disconnected kinetic events.