Detection of estrus in dairy cows by electrical measurements of vaginal mucus and by milk progesterone.

Electrical resistance (ohms) of mucus were analyzed in 20 postpartum Holstein cows by use of a probe inserted into the anterior vagina every other day for 30 days. Composite milk samples were taken on the same day, and progesterone was determined by radioimmunoassay. Cows were observed twice daily for standing estrus and reproductive organs palpated weekly per rectum (rectal palpation). Fifteen cows which were cycling showed increasing progesterone 6 to 7 days after the onset of estrus with values of 8.1 to 10.0 ng progesterone/ml milk on days 10 to 17. Concentrations had declined rapidly 2 days before onset of the next estrus. Progesterone in milk was affected by cow and by day of the cycle. Electrical resistance followed a similar cyclical pattern, but variability was large and only cows differed. The correlation between milk progesterone and mucus resistance was .22. Progesterone concentrations for four cows with follicular cysts fluctuated randomly with a mean of 2.6 ng/ml. Mean resistance of vaginal mucus was 44 omega for both cycling and cystic cows, indicating that a single measurement of electrical resistance every 2nd day was unreliable in distinguishing physiological states. One cow had high progesterone in milk on days 19 to 25 and was diagnosed pregnant by rectal palpation 3 wk later. Cows were not seen in estrus 28% of the time when milk progesterone and rectal palpation indicated they were in the follicular phase of the estrous cycle and were cycling.     

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.     

Duration of estrous cycles subsequent to two injections of prostaglandin F2 alpha given at a 14-day interval in nonlactating Holstein cows

Data from previous experiments in our laboratory indicated that estrous cycles following synchronization with two injections of prostaglandin F2 alpha often averaged more than 21 d. To investigate whether this could be an effect of prostaglandin F2 alpha, 54 mature heifers and nonlactating Holstein cows were used in a crossover experiment in which estrous cycles either occurred spontaneously or followed two injections of prostaglandin F2 alpha at a 14-d interval (induced estrus). One estrous cycle of each type was recorded for each animal. Blood samples were collected twice weekly for analysis of progesterone. Cattle were observed twice daily for estrus with the aid of tail chalking and an androgenized cow. Estrous cycles following induced estrus were longer than those following spontaneous estrus (24.2 vs. 22.7 d). Ninety-two percent of the cycles following spontaneous estrus were 17 to 25 d in duration compared with 71% of the cycles following induced estrus. If the first prostaglandin F2 alpha was given when progesterone was greater than 1 ng/ml, the estrous cycle following treatment was longer than if the first prostaglandin F2 alpha was given when progesterone was less than 1 ng/ml (26.0 vs. 22.6 d). This experiment demonstrates that two injections of prostaglandin F2 alpha increase the incidence of estrous cycles greater than 25 d. Stage of the estrous cycle at time of the first injection of prostaglandin F2 alpha may contribute to this effect.     

Reproductive performance of dairy cows with cystic ovaries following administration of Procystin

Eighty-five postpartum Holstein cows were randomly assigned to receive 0, 50, 100, or 250 micrograms of the gonadotropin-releasing hormone product, Procystin when follicular cysts were diagnosed by palpation per rectum. Accurate reproductive records were maintained, and milk samples were collected at the time of diagnoses for assay for progesterone. An additional 101 cows were injected with only the 100 micrograms dose of Procystin when cysts were identified. Data showed that days from treatment to first observed estrus decreased with increasing doses of Procystin with no advantage of 250 micrograms over 100 micrograms. Days open and conception rates were similar among the treatment groups. Cows with less than 1 ng/ml progesterone in their milk at the time of treatment returned to estrus sooner than cows with progesterone concentrations greater than 1 ng/ml. In addition, gonadotropin-releasing hormone administered to those cows with low progesterone at the time of treatment led to significantly increased progesterone concentrations by 7 and 14 d posttreatment. We conclude that although Procystin administration hastened estrus of cows with ovarian cysts, breeding practices on the farms did not lead to an improvement in reproductive efficiency of the cows that possessed cysts.     

Effect of progesterone on the expression of estrus at the first postpartum ovulation in dairy cattle.

Fifty-two lactating Holstein cows were randomly assigned to receive either a progesterone-releasing (2 g of progesterone) or a control-releasing intravaginal device (0 g of progesterone). Intravaginal devices were inserted on d 10 and removed on d 15 postpartum. Daily blood samples were collected from d 10 to 90 postpartum for subsequent determination of progesterone concentrations. Observations for estrus were conducted three times daily in a dirt paddock containing a testosterone-treated cow. Serum concentrations of progesterone in the progesterone-releasing intravaginal device group were elevated on d 11, 12, 13, 14, and 15 compared with those of the control group. The days to first post-partum ovulation were similar between the treated and control groups, respectively (30.6 vs. 30.5 d). Also similar was the proportion of cows expressing estrus at first, second, and third postpartum ovulations (9/27 vs. 3/24, 14/23 vs. 15/21, and 14/21 vs. 9/15, respectively), length of the first postpartum estrous cycle (17.9 vs. 18.3 d), and peak serum concentrations of progesterone during the first estrous cycle (3.5 vs. 2.9 ng/ml). These data indicate that administration of progesterone early postpartum did not increase the proportion of cows expressing estrus at the first ovulation.     

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.     

Synchronization of estrus in dairy cows with prostaglandin F2 alpha and estradiol benzoate

A field study using 322 lactating dairy cows in seven commercial and two university herds was conducted to determine if treatment with estradiol benzoate 40 to 48 h after treatment with prostaglandin F2 alpha would enhance synchronization of estrus. Estrogen treatment tended to increase the proportion of cows in estrus within 5 d (synchronized) after prostaglandin treatment (66.9% versus 58.9%). Of synchronized cows, a greater proportion treated with estrogen (66.9%) were in estrus on d 3 than those not receiving estrogen (48.2%). First service conception rate (31.9%) and interval to second service (35.6 d) were not affected by treatment with estrogen. Milk progesterone was measured in university herds. More cows with milk progesterone concentrations greater than or equal to 8 ng/ml were synchronized (75.4%) than those with less than 8 ng/ml (63.3%). Treatment with estrogen increased synchrony of cows with high progesterone (90.3%) more than prostaglandin alone (60.0%). Based on progesterone concentrations at breeding and 22 to 24 d later, estimated conception rate was 58.7%, and net conception rate based on palpation was 41.3%. Tighter synchrony of estrus can be achieved by using estradiol benzoate 40 to 48 h after prostaglandin. Concentrations of milk progesterone might predict success of treatment.     

Concentrations of progesterone in milk of cows administered an intravaginal progesterone insert

Milk from pregnant cows contains concentrations of progesterone (P4) considered safe for human consumption. The objective of this study was to determine if concentrations of P4 in milk during administration of an intravaginal progesterone insert (CIDR insert) are less than concentrations of P4 in milk associated with pregnancy. Results have implications for human use of milk from cows receiving CIDR inserts. Holstein cows (N = 64; > 40 and < 150 d after calving) were administered 25 mg of PGF2alpha i.m. (study d 0) and 20 cows detected in estrus from 2 to 4 d later were assigned randomly to either control (N = 10; no further treatment) or CIDR insert (N = 10; 1.38 g of P4) inserted on study d 17 (14 +/- 1 d after estrus) and removed 7 d later. Composite milk samples were collected contemporaneously from each of the 20 estrous cycling cows and from 10 pregnant cows (> or = 60 and < or = 220 d of gestation) twice daily from study d 17 to 27. Concentrations of P4 in defatted milk samples were quantified using a validated radioimmunoassay. Mean logs of areas under the curve of concentrations of P4 from the afternoon on study d 17 through the afternoon on study d 27 were 3.05 ng day/ml for control, 3.33 ng day/ml for CIDR insert, and 3.81 ng day/ml for pregnant cows. Therefore, increased P4 due to pregnancy was 0.76 ng day/ml (3.81-3.05), whereas the increase in P4 due to CIDR insert was only 0.28 ng day/ml (3.33-3.05). Applying a 95% confidence interval to 0.28 ng day/ml provided an upper value of 0.70 ng day/ml, lower than the increase due to pregnancy. Because milk from pregnant cows is considered safe for human consumption, it follows that milk from cows administered CIDR inserts should also be considered safe, based on concentrations of P4.     

Ovarian activity of Holstein and Jersey cows of diverse transmitting abilities for milk

The relationship between ovarian activity and milk yield was studied in 35 daughters of 24 Holstein sires and 17 daughters of 14 Jersey sires in the same herd. Ovulations and length of estrous cycles were determined by progesterone concentration in postmilking strippings three times per week, by weekly palpation per rectum, and by twice daily estrus detection. Transmitting abilities were for 4% fat-corrected milk of cows and their sires. Yields of 4% fat-corrected milk were estimated for 60, 90, 120, and 305 days in lactation. Postpartum intervals to first ovulation averaged 22 and 20 days for Holsteins and Jerseys, but the interval to first standing estrus was shorter for Jersey than for Holstein. Postpartum intervals to each of the first three ovulations and length of estrous cycles were unrelated to actual yield or transmitting ability for yield of 4% fat-corrected milk in either breed. The percentage of cows observed in standing estrus at each of the first three ovulations increased from 23 to 43%.     

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.     

Reproductive performance of grazing dairy cows following presynchronization and resynchronization protocols

Objectives were to compare the effect of presynchronization and resynchronization methods on fertility responses of grazing dairy cows at first and second artificial insemination (AI) and pregnancy rate during the entire breeding season. Lactating dairy cows (n = 1,263) in 2 seasonal grazing farms were blocked, within farm, by parity, breed and days in milk. Within each block, cows were randomly assigned to 1 of 4 treatments arranged as a 2 × 2 factorial with 2 presynchronization and 2 resynchronization treatments. Cows had their estrous cycles presynchronized with either a PGF_2α-based program (Presynch) consisting of 2 injections of PGF_2α administered 14 d apart and starting the timed AI protocol 11 d later, or with a PGF_2α-GnRH-based presynchronization program (G6G) consisting of an injection of PGF_2α, followed 3 d later by an injection of GnRH and starting the timed AI protocol 6 d later. All cows received the first insemination on the same day, which was considered study d 0 and also d 0 of the breeding season. All cows received the 5-d timed AI protocol that consisted of GnRH on d −8, PGF_2α on d −3 and −2, and GnRH + timed AI on d 0. Blood was sampled and analyzed for progesterone on d −8. On d 12, cows in each presynchronization treatment either remained as untreated controls (RCON) or received a controlled internal drug-release (CIDR) insert containing progesterone for 7 d (RCIDR). Estrus was observed daily starting on d 19 and cows in estrus were inseminated on the same day. On d 35, bulls were placed with the cows for an additional 65 d, completing a 100-d breeding season. Holstein cows were less likely to have progesterone ≥1 ng/mL on d −8, and had less expression of estrus and pregnancy per AI (P/AI), which resulted in a slower rate of pregnancy and a smaller proportion of pregnancy at the end of the study than did Jersey or crossbred cows. In addition, body condition, days in milk, and plasma progesterone concentration at the first GnRH injection of the timed AI protocol had marked effects on the reproductive performance of lactating grazing dairy cows. A greater proportion of G6G cows had progesterone ≥1ng/mL at the first GnRH injection of the timed AI protocol compared with Presynch cows (82.0 vs. 74.3%). Presynchronization treatment did not influence P/AI, but cows in G6G had increased risk of pregnancy loss between d 30 and 65 after the first AI (12.9 vs. 8.1%). Nevertheless, an interaction between presynchronization and ovarian status was observed, and cows initiating the timed AI with progesterone ≥1 ng/mL had greater P/AI when previously treated with Presynch than G6G. On the other hand, G6G benefited P/AI of cows initiating the timed AI with progesterone <1 ng/mL. Resynchronization with RCIDR altered the pattern of return to estrus, but it did not increase the rate of re-insemination and decreased the proportion of pregnant cows at the end of the 100-d breeding period (80.6 vs. 84.4%).     

Blood and milk progesterone in pregnant and nonpregnant buffalo

Changes in progesterone concentration of blood and milk were measured by radioimmunoassay in 10 Murrah buffalo up to 40 days after insemination. Progesterone concentration in blood plasma at estrus was .1 ng/ml which rose to a peak of 3.6 ng/ml on day 13. It continued to increase in animals that conceived but dropped to .6 ng/ml on 3 days before next estrus in those that failed to conceive. The average concentration of progesterone in milk was .5 ng/ml at estrus; it increased to 18 ng/ml on day 15, and thereafter it declined to 4.4 ng/ml 3 days preceding next estrus in nonpregnant animals. In pregnant animals, it was maintained and elevated further to 24.8 ng/ml on day 37. Progesterone in milk was four to five times higher than in blood plasma.     

Changes of luteinizing hormone and progesterone for dairy cows after gonadotropin-releasing hormone at first postpartum breeding

Gonadotropin-releasing hormone administered at breeding enhances fertility of dairy cows, so a study was designed to evaluate the mechanism for enhanced fertility following administration of gonadotropin-releasing hormone at first postpartum breeding. Twenty-four cows were assigned randomly to one of two treatments, 100 micrograms of gonadotropin-releasing hormone intramuscular or saline vehicle intramuscular at insemination. Blood samples for luteinizing hormone assay were taken at 2-h intervals prior to breeding and .5-h intervals for 3 h after insemination. Composite morning milk samples for progesterone assay were collected for 30 days after insemination or until next estrus. Cows given gonadotropin-releasing hormone had higher luteinizing hormone concentrations in blood serum following treatment than cows given saline, 13.2 versus 3.0 ng/ml. There was no relationship between luteinizing hormone and subsequent conception. Progesterone for cows that became pregnant was higher throughout sampling days. Mean progesterone concentrations were 4.6 versus 2.2 ng/ml in pregnant and nonpregnant cows during the first 4 days after insemination. Cows treated with gonadotropin-releasing hormone that conceived had higher progesterone than other cows, and that was evident at the first 4 days postbreeding.     

Interrelationships With Estrous Behavior and Conception in Dairy Cattle

A total of 199 inseminations and estrous periods of lactating cows and heifers of breeding age were used to assess behavioral, productive, environmental, and hormonal events at first observation of estrus and those 12 h later with concurrent breeding efficiencies. Cattle were observed twice daily for estrus, and blood samples were collected at the initial observation of estrus (0 h) and at 12 h. Fifty-one percent of the cattle first were detected in estrus in the morning. Morning estrous activity was greater (11.4 mounts/h) than that first observed in the evening (7.6). Mean estrous activity declined from 9.4 mounts/h at 0 h to 1.6 mounts/h at 12 h. Older cows exhibited more mounting activity than younger animals at 0 h. Barn housed cattle exhibited more mounts/h during detection of estrus (11.2) than cattle housed primarily in free stalls (6.5) or pasture (5.4). Early estrous activity (0 h) was not affected by estradiol, progesterone, or luteinizing hormone. However, estradiol and progesterone did exert an influence on activity seen at late estrus (12 h). Glucose and urea at 0 and 12 h were similar. There were no significant correlations of glucose or urea of plasma with estrous activity or conception. Feed intake and milk production from 3 days prior to 3 days postestrus did not change. Season affected estrous activity and conception. Highest conception was associated with low progesterone at 12 h.     

Glucocorticoids in mammary secretions and blood serum during reproduction and lactation and distributions of glucocorticoids, progesterone, and estrogens in fractions of milk

Concentrations of glucocorticoids were measured in milk and blood serum during the estrous cycle, prepartum, parturition, postpartum, and early and late lactation. Glucocorticoids in milk did not change during the estrous cycle, averaging .35 ng/ml whereas they averaged .50 ng/ml prepartum, 3.08 ng/ml at parturition, and .50 ng/ml 1 wk postpartum. Glucocorticoids in milk declined from .59 ng/ml to .25 ng/ml as lactation advanced from 1 to 10 mo. Concentrations of glucocorticoids in blood serum were approximately 8 to 10 ng/ml during all reproductive states. There was no positive relationship between percentage of milk fat and concentrations of glucocorticoids in milk. Cortisol was the predominant glucocorticoid in serum; in milk corticosterone concentrations exceeded those of cortisol. Glucocorticoids, being more polar, had higher affinity for the nonlipid portion of milk; in contrast, progesterone, estradiol 17beta, and estrone were located predominantly in the lipid fraction of milk.     

Management factors affecting reproductive performance of dairy cows in the northeastern United States

Dairy herds (476) in seven states in the northeastern United States were surveyed to determine effects of various management factors on reproductive performance. Error of estrus detection (greater than 1 ng/ml of milk progesterone) on the day of service was 5.1% for 4558 cows but was as high as 60% in some herds. Error was not affected by herd size but was greater in freestall (6.8%) than in conventional (5.2%) housing. "Standing" and "riding other cows" were the most accurate signs of estrus. Of cows in or near estrus when serviced, 28.1% were open 3 wk later, 12.9% were probably open, and 59% were probably pregnant based on analysis of milk progesterone. Conception rate, not affected by herd size or housing type, was greater for cows in estrus during the morning and serviced the same afternoon (52.2%) than for cows observed in the afternoon and serviced the next morning (47.1%). Fifty-five percent of cows open to first service were serviced again within 3 days of expected return to estrus. Days to first service and conception rates were correlated positively. Milk progesterone concentration 21 to 24 days after service predicted a cow will calve with 88.6% accuracy and that she will not with 93.9% accuracy. Veterinarians predicted a calf with 92.5% accuracy by rectal palpation.     

Changes of aldosterone in blood serum of dairy cattle during estrous cycle

Blood samples were taken daily from five lactating cows and five open heifers beginning with the 1st day postestrus and continuing until the animal was observed in standing estrus. Means of aldosterone in blood serum were 102.6 +/- 5.1 pg/ml for lactating cows and 94.1 +/- 5.1 pg/ml for open heifers. Age and lactation did not appear to be major factors affecting circulating aldosterone in the bovine species. Effect of day of the estrous cycle on aldosterone was quadratic. Concentrations of aldosterone for lactating cows started at 92.3 pg/ml on day 1 of the estrous cycle, peaked at 125.5 pg/ml on day 8, then declined steadily to a low of 43.2 pg/ml on day 20. There was no significant interaction of group by day. During the estrous cycle aldosterone appeared to be related to development of corpus luteum and progesterone. Sodium in blood serum was higher during estrus for lactating cows than open heifers for linear, quadratic, and cubic effects. Although not correlated with aldosterone, sodium exhibited similar cyclic patterns throughout the estrous cycle. Responses between sodium and aldosterone appeared delayed during estrus.     

Embryonic Mortality in Dairy Cows Estimated by Nonreturns to Service,Estrus, and Cyclic Milk Progesterone Patterns

Cyclic patterns of progesterone in milk were studied in 262 Holstein cows. Milk progesterone data were obtained from 153 cows for at least 75 days after conception as indicated by continuous high progesterone concentrations in milk for 28 days or more following artificial insemination. Cycling was reinitiated in 11 of these cows between 28 and 75 days after breeding, for an estimated 7.2% rate of embryo-fetal mortality. This was considerably less than a corresponding rate of 22.7% estimated for this herd by the delayed returns to estrus (28 to 75 days). In 350,180 cows inseminated with semen from Holstein bulls the estimated embryo-fetal mortality by the same delayed return to service method was 12.5%. Biases in this latter method of estimation are discussed.     

Detection of Estrus by Three Methods

Estrus was detected by three methods in 88 postpartum Holstein cows housed in tie-stalls and allowed free activity 1 to 2 h daily in separate exercise lots. Cows in each of three groups were observed visually for standing estrus by herdsmen. Cows (31) in Group I were observed only visually and served as controls, cows (33) in Group II were fitted with MateMaster rump-mounted detectors, and cows (24) in Group III were exposed to a testosterone-treated marking heifer. Cows with less than 1 ng/ml progesterone in serum on the day of detection and greater than 1 ng/ml 10 days later were designated as accurately detected in estrus. Accuracies of detections for cows in Groups I, II, and III were 68, 66, and 79%. Efficiencies of detections were the total observed estrous periods divided by the total expected estrous periods and were 51, 51, and 52%. In addition, overall accuracy, a mathematical product of individual accuracy and efficiency, represented the best measure of a detection method. Overall accuracies in Groups I, II, and III were 35, 34, and 41%.     

Activity monitoring and an enzyme immunoassay for milk progesterone to aid in the detection of estrus.

The accuracy and efficiency of estrus detection using an electronic activity monitor tag in conjunction with an enzyme immunoassay for milk progesterone were studied during 55 observed estrous periods in 37 cows. At approximately 30 d postpartum, cows were equipped with an activity tag and visually observed for estrus, an activity tag with a flashing light-emitting diode, or both twice daily. Milk progesterone concentrations were determined from cows observed in estrus or with an activated tag. Mean daily activity was greater on the day of estrus than during any of the 3 d preceding or 3 d following estrus. Functioning activity tags correctly identified 55% of all visually observed estrous periods that coincided with low milk progesterone levels. The overall accuracy of a flag by the tag for identifying true estrus was 21%. The enzyme immunoassay for milk progesterone was in agreement with 98% of all visually observed estrous periods and false flags committed by the tags. Although the activity tags detected some cows in estrus, a more durable and reliable tag must be developed before it is of practical value to dairy producers.