Determinants of estrous behavior in lactating Holstein cows

The objective was to determine factors that affect the expression of estrus. Thirteen lactating Holstein cows were ovariectomized about 4 to 6 wk postpartum and then challenged repeatedly with progesterone and estradiol benzoate to induce estrus six times during the postpartum period. Each challenge included 5 d when the cow was primed with progesterone through insertion of a progesterone-impregnated, foam rubber pessary. Estradiol benzoate (1 mg) was injected intramuscularly 36 h after removal of the pessary. Groups of two to three cows each began the experiment at 3-mo intervals to avoid confounding treated simultaneously. Observations for estrous behavior were at 8-h intervals following each challenge. A minimum of three sexually active cows were always observed together to avoid differences in estrous behavior caused by having too few sexually active animals in the group. Observations for estrous behavior were at 8-h intervals following each challenge. During each observation, cows were observed for 30 min on dirt and for 30 min on concrete. Standing behavior was not influenced by postpartum interval, season of year, or milk yield. Mounting behavior increased from the first to the sixth postpartum challenge, but it was not affected by season of year or milk yield. Duration of estrus, mounting activity, and standing activity were greater on dirt than on concrete. These results indicate that the surface on which cows were observed had a profound effect on sexual behavior; however, postpartum interval, season of year, and milk yield were of minor importance.     

Comparative feed efficiency of Holstein and Jersey cows

Feed efficiency measures, gross energy (solids-corrected milk/estimated energy intake), N (milk N/N intake) utilization, and apparent digestibilities of ration dry matter and N were calculated for 34 daughters of 21 Holstein and 29 daughters of 18 Jersey sires in first and second trimesters of lactation. Cows were studied in N balance trials and were fed ad libitum corn silage-based, complete rations formulated to meet average nutrient requirements. The objective was to determine whether these breeds differ in rates of converting dietary protein or energy to milk protein or milk energy. Holstein and Jersey did not differ for digestibility of ration dry matter in either trimester of lactation. They also converted dietary protein to milk protein at similar rates. Energy efficiencies of Holstein and Jersey did not differ in first trimester, but Holstein exceeded Jersey in second trimester. Because intake of dry matter by Jersey increased about 4% from first to second trimester (opposite of Holstein) and N balance was higher than for Holstein, Jersey cows may have been repleting more adipose tissue. We suggest no comparative advantage for Jersey in spite of higher ratios of milk to body weight and feed intake less than Holstein.     

Gonadotropin-releasing hormone and prostaglandin F2 alpha for postpartum dairy cows: estrous, ovulation, and fertility traits

A study of 234 Holstein cows was conducted to determine if early postpartum treatments of gonadotropin-releasing hormone, prostaglandin F2 alpha, or both would alter frequency and occurrence of estrus and ovulation as well as subsequent fertility. Cows in groups 1 and 2 received gonadotropin-releasing hormone (200 micrograms) between 10 and 14 d postpartum, and cows in groups 3 and 4 received saline. Ten days later, cows in groups 2 and 3 received prostaglandin F2 alpha (25 mg), and cows in groups 1 and 4 received saline. Treatment with gonadotropin-releasing hormone reduced intervals to first ovulation and first detected estrus as well as increasing the proportion of cows with three or more ovulations before first service from 57% for saline-treated controls to 83%. Treatment with prostaglandin F2 alpha reduced intervals to second and third ovulation and shortened the first estrous cycle. Treatments for cows in groups 1 and 3 increased the proportion of cows having estrous cycles of normal duration. Interval from calving to conception was reduced by 43 to 48 d for cows with an abnormal puerperium treated with either gonadotropin-releasing hormone or prostaglandin F2 alpha compared with controls (group 4) and by 27 to 29 d overall in all cows in groups 1 and 3. Cows with normal or abnormal puerperium in groups 1 and 3 required 26 to 41% fewer services per conception than controls. We conclude that treatments of gonadotropin releasing hormone or prostaglandin F2 alpha, but not the treatment combination, improved fertility of dairy cows, especially those that experienced puerperal problems.     

Increased milk production versus reproductive and energy status of high producing dairy cows

Two groups of 10 Holstein cows were chosen by pairs from a 20-yr genetic selection project that used either breed average or breed high sires chosen only for Predicted Differences in milk production. Milk production (305-d mature equivalent) was 10,814 kg and 6912 kg for the high and average groups of cows. Days to first visual estrus and number of ovulations before first visual estrus were greater for the high versus the average group (66 vs. 43 d and 1.6 vs. .7 ovulations). No differences were significant between groups for the interval from parturition to uterine involution or for days to first ovulation. Energy balance was less for the high group during wk 1, 2, 10, and 11. Plasma glucose concentration was lowest during wk 2 for both groups, and nonesterified fatty acids and beta-hydroxybutyrate were greatest for both groups during wk 1 and 2. Liver glycogen content was lower at d 15 postpartum for the high group, and liver triglyceride content was greater on d 30 for the high group. The data for reproductive functions support the concept that high milk production is antagonistic to expression of estrous behavior but not to reactivation of ovarian function.     

Effect of additional cobalt, copper, manganese, and zinc on reproduction and milk yield of lactating dairy cows receiving bovine somatotropin.

The objective of this study was to determine whether organically complexed Co, Cu, Mn, and Zn would improve the reproductive performance and milk and milk component production in lactating dairy cows that began receiving bovine somatotropin in the ninth week of lactation. Holstein (n = 50) and Jersey (n = 10) cows were blocked by breed, lactation number, and incidence of retained fetal membranes. Two diets assigned within blocks and fed from parturition until 154 d of lactation were control or control supplemented daily with 26 mg of Co as Co glucoheptonate, 125 mg of Cu as Cu-Lys, 199 mg of Mn as Mn-Met, and 359 mg of Zn as Zn-Met. Cows were fitted with electronic pressure-sensing devices in the second week of lactation for detection of estrus. Ovarian structures were determined via transrectal ultrasonography at 7-d intervals from parturition until observation of the first corpus luteum. Blood samples were taken at 7-d intervals and analyzed for plasma concentrations of progesterone, insulin, and urea nitrogen. Onset of luteal activity was identified by progesterone concentrations > or = 1 ng/ml. Retained fetal membranes increased days to first estrus (detected via electronic estrous detection), first luteal activity, and first corpus luteum in control cows but not in supplemented cows. Days to first observed estrus were greater for control cows than for supplemented cows. Days to first service, days open, days from first service to conception, services per conception, milk yield, milk components, and somatic cell counts were similar for control and supplemented cows. Supplementation with complexed trace minerals effectively reduced days to first estrus.     

Postpartum ovarian follicular dynamics and estrous activity in lactating dairy cows

The objectives of this study were to characterize early postpartum follicular dynamics in dairy cows in relation to their estrual activity and subsequent reproductive performance using 50 (26 primiparous and 24 multiparous) lactating Holstein cows. Ovaries and uterine horns of postpartum lactating cows were examined by ultrasonography 3 times weekly and continued until first services occurred after a 45-d voluntary waiting period. No differences were detected in fertility between primiparous and multiparous cows. In 40 of 50 cows, first postpartum ovulation was observed within 4 follicular waves, and the follicular wave patterns and ovarian cycles in most cows returned to normal as in cattle having normal estrous cycles after the second postpartum ovulation. Cows with the longest intervals from calving to first ovulation produced the most milk and also had prolonged intervals to first estrous activity. Differences in follicular dynamics before first ovulation altered intervals to first estrus, first service, and uterine involution, but these differences did not affect pregnancy rate, number of services, and days open. First postpartum insemination after 3 follicular waves tended to have greater pregnancy rates than those after 2 follicular waves. First inseminations at first estrus could produce greater pregnancy rates than those at subsequent periods of estrus.     

Growth, reproduction, and lactation in somatic cell cloned cows with short telomeres

We previously showed that telomere lengths of 10 somatic cell cloned cows were significantly shorter than normal. In this study, we investigated growth, reproduction, and lactation in these animals to determine if shortened telomeres have any effect on these characteristics. Six Holstein and 4 Jersey cloned cows, derived from oviduct cells, were reared under general group feeding. Body weights were recorded from birth to 48 mo of age. A number of reproductive characteristics were screened during the prepubertal, postpubertal, and postpartum periods. After parturition, milk yields were recorded daily and percentages of milk fat, proteins, and solids-not-fat were measured at monthly intervals. These data were used to estimate production of milk components over a 305-d period. Overall, the cloned heifers exceeded standard growth rates for each breed. The cows were inseminated at the first estrus after they reached 450 d of age, and delivered normal calves except for one stillbirth in the Holstein group. They were inseminated at postpartum estrus to provide second and third parturitions and, again, these pregnancies were normal. Gestational periods and birth weights of the calves were both within the normal range. The average total milk yield per cow in Holstein group clones was less than that of the original cow, whereas Jersey group clones showed a higher average milk yield than the original cow. In both groups of cloned cows, inter-individual variation in milk production was relatively large; however, the coefficient of variation was less than 10%. Our results suggest that the cloned cows have normal growth, reproductive, and lactation characteristics, and thus normal productivity, despite having reduced telomere lengths.     

Factors affecting reproductive performance of dairy cows first inseminated after five weeks postpartum

Various factors influencing reproduction of 307 Holstein cows were evaluated. Sources of variation of several measures were estrous certainty, time of day of detected estrus and insemination, method of thawing semen, ease of cervix penetration, presence of clear mucus at insemination, service number, service sire, inseminator, age of cow, maximum ambient temperature on the service date, and milk yield. Interval to first service of older cows was longer than of younger cows and shorter for cows with higher milk yield. Conception rate was higher for inseminations before noon than after noon and tended to be lower at inseminations when milk yield was at the extremes (low or high). Rate of conception was similar for cows whether inseminated at early or later postpartum intervals. More services were required by cows that were inseminated when milk yield was at the extremes (low or high). Fewer days open were associated with fewer total services and younger age at conception. Cows inseminated with semen thawed in hot (65 degrees C, 7 to 10 s) or warm (35 degrees C, 30 s) water conceived earlier postpartum than did those inseminated with semen thawed in the cow. Submitting cows for insemination after detection of standing estrus, mounting activity, or less certain symptoms of estrus had no adverse effect on fertility or days open.     

Herdlife Lactation Yield,Herdlife, and Survival of Holstein and Ayrshire-Based Lines of Dairy Cattle

Length of herdlife, lifetime milk and component yields, daily milk and component yields, and survival rates were assessed from multiple lactation records of 927 cows in a Holstein line and an Ayrshire-based synthetic line. Holstein cows were progeny of random matings to either Holstein untested sires or progeny-tested Canadian and American Holstein sires. Ayrshire cows were randomly mated to Ayrshire untested sires or Canadian and American Ayrshire, Finnish Ayrshire, Norwegian Red, or Brown Swiss progeny-tested sires to produce the foundation females in the Ayrshire-based synthetic line.The Holstein cattle produced more milk and protein per day of milking herdlife or total herdlife than cows in the Ayrshire-based line and proportionately more survived to initiate second lactations. Daughters of the progeny-tested American and Canadian Holstein sires had significantly higher lifetime and daily milk, protein, and fat yields than daughters of young untested Holstein sires whereas lifetime yields of daughters of untested and progeny-tested Ayrshire sires were equivalent.     

Implementation of a routine genetic evaluation for longevity based on survival analysis techniques in dairy cattle populations in Switzerland

Genetic evaluation of sires for functional longevity of their daughters based on survival analysis has been implemented in the populations of Braunvieh, Simmental, and Holstein cattle in Switzerland. A Weibull mixed sire-maternal grandsire survival model was used to estimate breeding values of sires with data on cows that calved since April 1, 1980. Data on Braunvieh and Simmental cows included about 1.1 million records, data on Holstein cows comprised about 220,000 records. Data contained approximately 20 to 24% right-censored records and 6 to 9% left-truncated records. Besides the random sire and maternal grandsire effects, the model included effects of herd-year-season, age at first calving, parity, stage of lactation, alpine pasturing (Braunvieh and Simmental), and relative milk yield and relative fat and protein percentage within herd to account for culling for production. Heritability of functional longevity, estimated on a subset of data including approximately 150,000 animals, were 0.181, 0.198, and 0.184 for Braunvieh, Simmental, and Holstein, respectively. Breeding values were estimated for all sires with at least six daughters or three granddaughters in the data. Breeding values of sires are expressed in months of functional productive life and published in sire catalogs along with breeding values for production traits.     

Changes throughout estrous cycles of variables that might indicate estrus in dairy cows

Two experiments examined physiological changes throughout estrous cycles of Holstein cows. In experiment one, changes were characterized for physical activity, vaginal pH, vaginal temperature, milk yield, heart rate, and concentration in blood plasma of progesterone and 13,14-dihydro-15-keto-prostaglandin F2 alpha. Variables were measured daily in 11 cows through a total of 28 estrous cycles. Cows in estrus at least once postpartum were used. Observations for estrus were at least twice daily. Variables changed significantly with day of estrous cycle. Changes of progesterone, physical activity, and 13,14-dihydro-15-keto-prostaglandin F2 alpha were more marked around estrus than were changes of the other variables. Peaks of physical activity coincided with estrus in 75% of the cases. Experiment two was to determine if increased physical activity, measured with pedometers, coincided with estrus in commercial dairy herds. Cows on 14 farms in New York were fitted with mechanical pedometers, and the pedometers were read at each milking. Readings from 3 days before until 3 days after estrus were used from 55 estrous periods identified by the producers. Physical activity was maximal on day of estrus in 73% of the estrous periods. Increased physical activity of commercial dairy cows agreed well with the producers' diagnoses of estrus. Pedometers could be a valuable component of an estrous detection program.     

Effectiveness of rump-mounted devices and androgenized females for detection of estrus in dairy cattle

Detection of estrus was evaluated in 1124 estrous cycles for the effectiveness of rump-mounted devices and androgenized females equipped with chin-ball markers. Estrous number, location, type of detection method (compared with visual observation of estrus), and milk progesterone influenced standing behavior. Rates of disagreement with visual observation of estrus were 13.2, 17.0, and 18.5% for Kamar, Hot Flash, and the two devices used simultaneously. Frequency of observed standing behavior was greater with fourth and subsequent estrous periods. Percentage of high milk progesterone during suspected estrus for cows inseminated was 3.8% for controls, 11.0% for Kamar cattle, 18.5% for Hot Flash cattle, and 12.6% when both devices were used. Androgenized cattle equipped with chin-ball markers marked 47% of the cattle observed in estrus but no cows with high milk progesterone. Combination of rump-mounted devices and androgenized females increased the percentage of estrous detection by 1.3 to 6.2% compared with device or androgenized female used alone. Loss rates of rump-mounted devices exceeded 40%. High milk progesterone at estrus was associated with lower fertility.     

Production, reproduction, health, and growth traits in backcross Holstein × Jersey cows and their Holstein contemporaries

A total of 648 purebred Holstein and 319 backcross Holstein × Jersey dairy cattle were compared for production, reproduction, health, linear type, and growth traits. Animals were born between 2003 and 2009 and were housed in the University of Wisconsin–Madison Integrated Dairy Facility. All animals had Holstein dams; lactating dams were mated to unproven Holstein sires to produce purebred (control) Holsteins or to unproven F₁ Jersey × Holstein crossbred sires to produce backcross animals, whereas nulliparous dams were mated to proven Holstein sires to produce purebred (other) Holsteins. Traits were analyzed using mixed linear models with effects of season of birth, age of dam, sire, birth year of sire, days in milk, lactation, and linear type score evaluator. Control Holsteins had greater 305-d milk yield (12,645 vs. 11,456 kg), 305-d mature equivalent milk yield (13,420 vs. 12,180 kg), peak daily milk yield (49.5 vs. 46.4 kg), total lactation milk yield (11,556 vs. 10,796 kg), and daily fat-corrected milk yield (43 vs. 40 kg) compared with backcrosses. Days open and services per conception as a heifer or cow did not differ between control Holsteins, other Holsteins, or backcrosses. The proportion of first-parity births that required assistance was less in control Holsteins than in backcross cows (3.7 vs. 11.2%). The incidence of scours or respiratory problems in calves did not differ between control Holsteins, other Holsteins, and backcrosses, nor did the incidence of mastitis, injury, or feet problems. Control Holstein heifers were heavier (629 vs. 557 kg), with greater hip height (145 vs. 139 cm), body length (167 vs. 163 cm), heart girth (205 vs. 198 cm), and hip width (54 vs. 53 cm) at 22 mo of age. On a 50-point scale for linear type traits, Holsteins were larger in stature compared with backcrosses (41 vs. 28), had wider rumps (37 vs. 33), and wider rear udders (34 vs. 32). Results of this study suggest that backcross Holstein × Jersey cattle have decreased production but fail to demonstrate an advantage in health and reproduction compared with purebred Holsteins.     

Use of Mount Detectors Plus Chalk as an Estrous Detection Aid for Dairy Cattle

Two experiments were conducted to determine effectiveness of mount detectors plus chalk as an estrous detection aid. Of 33 Holstein cows receiving more than one mount during continuous observations, three separate 30-min detection periods by a herdsman detected 20 (60.6%) of these cows in standing estrus. Use of either mount detectors, chalk, or mount detectors plus chalk indicated 31 (93.9%) of these cows in estrus. There were no false positives with estrous detection by the herdsman. Mount detectors and chalk had 28 and 38 false positives, respectively, compared with three false positives with the mount detectors plus chalk. Cows on three Holstein farms were assigned to treatments as 1) control, 2) cows with mount detectors, 3) cows with chalk, or 4) cows with mount detectors plus chalk. Based on 21-d estrous periods, treatments 1 to 4 resulted in 42.5, 56.6, 44.0, and 63.8% of estrous periods detected, respectively. Overall, cows not becoming pregnant on treatments 1 to 4 were 32.0, 23.0, 20.3, and 12.0%, respectively. Both cows with mount detectors and cows with mount detectors plus chalk had fewer days to second estrus than control cows and cows with chalk. Cows with mount detectors plus chalk had fewer days to conception than controls and cows with mount detectors. Cows with chalk had greatest days to conception.     

Variance-covariance components associated with trimester yields of milk and fat and multiple trait sire evaluation for trimester yields

Genotypic and phenotypic variances and covariances for milk and fat yields of three equally-spaced intervals in first lactation, days 1 to 90, 91 to 180, and 181 to 270, were estimated from 26,523 records of initial progeny-test daughters of 2,086 Holstein sires. Henderson's method 3 was used with a model that included fixed herd-years, fixed age-month of freshening, and random sire and residual effects. Heritabilities for milk yields were .21, .21, and .13 for the first, second, and third 90 days in lactation and for fat yields were .19, .16, and .10. Heritabilities for milk and fat yields over the entire 270 days postpartum were .22 and .22. Genetic correlations among partial yields were high and ranged from .74 to .99 for milk and .86 to .99 for fat. Genotypic and phenotypic variances were used to predict breeding values for trimester milk yield by multiple trait, mixed model procedures for 3,797 Holstein artificial insemination sires from 283,900 first-lactation milk records of their daughters during the first 90, second 90, and third 90 days in lactation. Correlations among proofs for the three trimesters ranged from .85 to .94. Correlation between the sum of three trimester multiple trait proofs and proof for milk yield for 270 days of lactation from single trait analysis was .99. Correlations among proofs indicate sires may rank differently for milk yield in the three periods.     

Relationship of the estrous cycle to milk composition.

The relationship of the estrous cycle to milk composition was examined in 15 Holstein cows in first lactation which were approximately 45 days postpartum. Composite milk samples were collected twice daily and analyzed for progesterone, fat, protein, total solids, sodium, potassium, magnesium, calcium, and somatic cells. Milk progesterone was minimal (less than ng/ml) from day -1 to day 2 (day 0 = estrus) of the estrous cycle and then increased at a relatively constant rate through day 15. Milk yield and sodium and magnesium concentrations of the milk varied with days of the estrous cycle. Mean milk yield was highest and sodium and magnesium concentrations were lowest on day 1. However, sodium concentration of the milk was the only component that varied significantly during the 3 days centered on estrus (days -1, 0, and 1). No other milk component changed significantly during the estrous cycle. Although milk yield and composition varied during the estrous cycle, none of these components appeared to be a practical indicator of estrus.     

The association between milk urea nitrogen and DHI production variables in western commercial dairy herds

A retrospective observational study was conducted using data from Dairy Herd Improvement monthly tests to investigate the association between milk urea nitrogen (MUN) concentration and milk yield, milk protein, milk fat percentage, SCC, and parity for commercial Holstein and Jersey herds in Utah, Idaho, and Montana. Mean MUN for Holstein cows was 15.5 mg/ dl (5.5 mmol/L) MUN and 14.1 mg/dl (5.0 mmol/L) for Jersey cows. Mean MUN, categorized by 30-d increments of days in milk (DIM), paralleled changes in milk values and followed a curvilinear shape. For Holstein cows, concentrations of MUN were different among lactation groups 1, 2, and 3+ for the first 90 DIM for Holsteins. Overall, concentrations of MUN were lower during for the first 30 DIM compared with all other DIM categories for both Holstein and Jersey cows. Multivariate regression models of MUN by milk protein showed that as the milk protein percentage increased, MUN concentration decreased; however, models for Jersey cows showed that MUN did not decrease significantly until above 3.4% milk protein. Milk fat percentage also decreased as MUN increased, but by only 1 mg/dl MUN over the range of 2.2 to 5.8% milk fat. Somatic cell count showed a negative relationship with MUN. Holstein cows with milk protein percentage >3.2% had lower MUN compared with cows having milk protein <3.2% for milk yields from 27.3 to 54.5 kg/d and lower than cows having a milk protein <3.0% for milk yield of 54.5 to 63.6 kg/d. In Jersey cows, MUN concentrations were not different among milk protein percentage categorized by milk yield. This study found that MUN was inversely associated with milk protein percentage and paralleled change in milk yield over time.     

Short communication: Characterization of early postpartum estrous behavior and ovulation in lactating dairy cows using radiotelemetry

The objective of this study was to describe early postpartum estrous behavior and ovulation in lactating dairy cows using radiotelemetry. Cows (n=50) were continuously monitored for behavioral estrus with a radiotelemetric system, HeatWatch II (CowChips LLC, Manalapan, NJ), from d 14 to approximately d 49 postpartum. Blood collection for analysis of progesterone and ovarian ultrasonography were performed once weekly starting on d 14. First ovulation was associated with behavioral estrus in 5 cows and occurred at 28.2±10.8d (mean±SD; range 17 to 40d). The average duration of estrus was 6.0±4.9h (range 3 to 12.2h), and the mean number of standing events was 18.4±8.9 (range 4 to 26). Based on progesterone concentrations of ≥1ng/mL, estimated first postpartum ovulation occurred at 25.1±10.4d (range 10 to 49d) for 38 animals without evidence of behavioral estrus. The interval to estimated first ovulation without behavioral estrus was not different from the interval to first ovulation associated with behavioral estrus. Level of milk production and body condition score loss did not affect the interval to estimated first ovulation without estrus or first ovulation associated with estrus. Six animals did not show evidence of ovulation based on progesterone concentration, whereas 1 cow showed evidence of estrous behavior on the day before removal from the study. The majority of first postpartum ovulations (38/43; 88.4%) were not associated with behavioral estrus.     

Estimates of genetic trend in an artificial insemination progeny test program and their association with herd characteristics.

Individual lactation records from Holstein cows in 3449 herds participating in an AI stud's young sire sampling program from 1971 to 1987 were used to characterize the sampling program and to estimate genetic merit and trend. Average genetic merit of cows in sampling program herds was consistently superior to the average genetic merit of cows in the US population. Genetic trend of sires of first-crop cows was 58 kg of milk and 1.5 kg of fat/yr from 1971 to 1978 and 176 kg of milk and 5.5 kg of fat/yr from 1979 to 1987. The average genetic merit of sires of first-crop cows born after 1983 was equivalent to or exceeded the genetic level of sires of other cows in the herd. Within-herd-year means and standard deviations of yield, genetic evaluation, and management traits (herd-year characteristics) were computed for a subset of 341 herds contributing first-crop daughters for at least 10 yr. The average of each herd-year characteristic during 10 or more years was used to predict within-herd genetic trend. Herd characteristics explained up to 51% of differences in within-herd genetic trends. Average sire genetic merit of daughters other than first-crop daughters accounted for up to 80% of the explained differences. Other herd characteristics suggested that herds with larger within-herd standard deviation milk yields, a larger number of young sires represented, younger cows, and greater percentage of cows sired by AI sires made greater genetic improvement. Results indicated that the average genetic merit of cows and the rate of within-herd genetic improvement are higher in herds that participate in a young sire sampling program.     

Relationship between level of milk production and multiple ovulations in lactating dairy cows

Our objective was to evaluate factors associated with spontaneous multiple ovulations in lactating dairy cows. Ovaries of cows [n = 267; >50 days in milk (DIM)] were evaluated weekly using ultrasound to determine spontaneous (i.e., no hormonal treatment) ovulation rate starting at 50 DIM and continuing until pregnancy diagnosis. Cows were fitted with a transmitter to record standing activity during estrus, and serum progesterone concentration was assessed weekly starting at wk 1 postpartum for all cows. Overall, 76 (28.5%) cows were anovular and 191 (71.5%) were ovular by 71 DIM. Incidence of anovulation was not associated with level of milk production but was associated with lower body condition. For anovular cows (n = 41) that spontaneously recovered, the multiple ovulation rate at first ovulation was 46.3%. For second and subsequent ovulations (n = 463), the level of milk production for 14 d preceding estrus was associated with increased ovulation rate. To illustrate, incidence of multiple ovulations was 1.6% (2/128), 16.9% (32/189), and 47.9% (70/146) for ovulations when cows were producing <35, 35 to <45, and ≥45 kg/d, respectively. Among cows for which estrous behavior was recorded, those with multiple ovulations (n = 48) had shorter duration of estrus (4.3 ± 0.7 vs. 9.9 ± 0.5 h) and higher production (47.2 ± 0.9 vs. 38.1 ± 0.5 kg/d) than cows with single ovulations (n = 237). Circulating concentrations of estradiol were lower (5.5 ± 0.3; n = 15 vs. 7.8 ± 0.4 pg/mL; n = 71) during periods of estrus with multiple ovulations despite a greater preovulatory follicular volume (4136 ± 123 vs. 3085 ± 110 mm³). Similarly, serum progesterone concentration 7 d after estrus was lower for cows with multiple than single ovulations (2.5 ± 0.3 vs. 3.2 ± 0.1 ng/mL) despite a greater luteal volume (8291 ± 516 vs. 6405 ± 158 mm³). In summary, the first spontaneous ovulation in anovular cows and a higher level of milk production for 14 d preceding estrus were associated with increased multiple ovulation rate. Additionally, cows with multiple ovulations had lower estradiol at estrus, a shorter duration of estrus, and lower progesterone at 7 d after estrus than cows with single ovulations.