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.     

Effect of supplemental beta-carotene on incidence and responsiveness of ovarian cysts to hormone treatment

Sixty-two multiparous and 35 primiparous Holstein cows were assigned randomly at 10 days postpartum to receive a ration with or without 300 mg beta-carotene/cow per day. Multiparous and primiparous cows were grouped separately and group-fed complete rations once daily. Incidence of ovarian cysts (26% by rectal palpation) was not affected by beta-carotene fed. Multiparous cows had greater incidence (39%) of ovarian cysts than primiparous cows (11%). Fifty-seven percent of cysts were classified follicular by rectal palpation. Progesterone concentration of milk also was used for diagnosis of type of cyst. Cows with ovarian cysts and with progesterone concentrations in milk less than 1 ng/ml were classified follicular, and those having concentration greater than 1 ng/ml were classified luteal. As determined by milk progesterone, rectal palpation was more accurate for diagnosis of luteal cysts than for diagnosis of follicular cysts. Progesterone concentrations of milk for animals with luteal and follicular cysts were 10.66 +/- 1.29 and .37 +/- .07 ng/ml. beta-Carotene did not affect response or days to respond to treatment with human chorionic gonadotropin or gonadotropin-releasing hormone. Supplemental beta-carotene was not beneficial for reducing incidence of ovarian cysts in cows receiving an adequate supply of beta-carotene in their diet.     

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.     

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.     

Gonadotropin-releasing hormone and conception of Holsteins

Effects of gonadotropin-releasing hormone (100 micrograms) and time of artificial insemination on fertility, were examined in lactating dairy cows at first, second, and third services. Inseminations were either soon after detected estrus (0 h) or 12 h later, and cows were given gonadotropin-releasing hormone or saline after inseminations. Conception at first service was not improved by hormone treatment. But conception rates at repeat services were improved by nearly 21% when cows were treated with gonadotropin-releasing hormone within 30 s after insemination. Inseminations soon after detection of estrus (0 h) followed by hormone treatment resulted in the highest conception rates for all services. Time of the insemination (0 versus 12 h) had no effect on conception. Administering gonadotropin-releasing hormone at repeat services should improve conception rate of lactating dairy cows.     

Controlling first service and calving interval by prostaglandin F2 alpha, gonadotropin-releasing hormone, and timed insemination

Our objective was to determine if calving intervals could be shortened and made less variable by using prostaglandin F2 alpha to control the occurrence of first services. Holstein cows (n = 348) were assigned at calving to four treatment groups. Control cows (n = 88) were inseminated at their first observed estrus after 40 d postpartum. Estrous cycles of the remaining cows were synchronized with prostaglandin F2 alpha to allow insemination (first services) 80 h after the second injection (n = 86), insemination at 80 h preceded by gonadotropin-releasing hormone at 72 h (n = 86), or insemination at 72 and 96 h (n = 88) after the second injection (51 to 57 d postpartum). By design, interval to first service was reduced to 57 d for treated cows (63 d for controls) and was less variable (12% of that for controls). Conception rate at first service was lower after timed inseminations than that of controls. Intervals to conception and subsequent calving were similar in all treated cows and controls. Of cows sampled, 23 of 176 (13%) failed to respond with luteolysis when progesterone in serum exceeded 1 ng/ml, and 26 of 176 (15%) had low concentrations of progesterone in serum and could not respond to prostaglandin F2 alpha. Poor response to timed inseminations may have occurred because only 72% of 176 cows sampled responded with luteolysis. When only cows observed in estrus were considered, conception rate approached that of controls (51%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of gonadotropin-releasing hormone and timing of insemination relative to estrus on pregnancy rates of dairy cattle at first service

The objective was to determine the influence of gonadotropin-releasing hormone on pregnancy rates of dairy cattle at first services, when both the timing of hormone injection and insemination were altered relative to the onset of estrus. Cows (n = 325) were assigned randomly to six groups making up a 2 X 2 X 2 incomplete factorial experiment; dose of GnRH (100 micrograms versus saline), timing [1 h (early) or 12 to 16 h (late) after first detected estrus] of AI, and timing of hormone injection (early versus late) were the three main effects. Cows were observed for estrus 4 times daily. Treatments and resulting pregnancy rates were: 1) hormone injection early plus AI early (35%), 2) hormone injection late plus AI early (34%), 3) saline injection early plus AI early (30%), 4) hormone injection late plus AI late (30%), 5) hormone injection early plus AI late (46%), and 6) saline injection late plus AI late (43%). Pregnancy rate in the first four groups (32%) was less than that in the latter two groups (44%). Concentrations of LH in serum were greater for cows given hormone or saline injections in early estrus than for cows injected with either hormone of saline during late estrus. Concentrations of LH in serum 2 h after GnRH were elevated above those of controls, whether GnRH was injected during early or late estrus. Neither concentrations of LH during estrus nor concentrations of progesterone 8 to 14 d after estrus explained the possible antifertility effect of GnRH given during late estrus.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Reproductive performance of dairy cows fed two concentrations of phosphorus

The objective of this study was to determine the effect of dietary P concentrations of 0.37 (recommended) or 0.57% (excess; dry matter basis) on reproductive performance. At calving, Holstein cows were randomly assigned to 1 of 2 dietary treatments (n = 134 for 0.37% P and n = 133 for 0.57% P). Cows were fitted with a radiotelemetric transmitter (50 d in milk [DIM]) to record mounting activity during estrus and bred to natural estrus from 50 to 100 DIM, then to synchronized estrus (Ovsynch protocol) after 100 DIM. Weekly ultrasonography was performed from 50 DIM until pregnancy was diagnosed (-30 d after artificial insemination). Pregnancy was confirmed approximately 60 d after artificial insemination (artificial insemination). Weekly blood samples were analyzed for progesterone concentrations. Days to first increase (>1 ng/ml) in progesterone, days to first estrus detected by radiotelemetry, days to first service detected by herd personnel, and conception rate at first service did not differ between the recommended and excess P groups, respectively. Similarly, conception rate at 30 d, days open, pregnancies lost from 30 to 60 d, multiple ovulation rate, and the incidence of anovulatory condition at 71 DIM did not differ between these groups. The mean duration of estrus was 8.7 +/- 0.5 and 8.7 +/- 0.7 h, and the average number of mounts per estrus was 7.4 +/- 0.5 and 7.8 +/- 0.5 for a total mounting time during estrus of 25.8 +/- 1.8 and 24.5 +/- 1.6 s for cows fed the recommended and excess P diet, respectively. Phosphorus treatment had no detectable effect on reproductive performance.     

Fertility of postpartum dairy cows after administration of gonadotropin-releasing hormone and prostaglandin F2 alpha: a field trial

Our objective was to examine further the potential profertility effects of gonadotropin-releasing hormone and prostaglandin F2 alpha in postpartum Holstein cows. Reproductive performance was monitored in 843 cows milked thrice daily. One group of cows (n = 218) was untreated, while three groups received either 100 micrograms gonadotropin-releasing hormone administered once between d 11 and 25 (n = 211); 25 mg prostaglandin F2 alpha given once between d 11 and 25 (n = 215); or 25 mg prostaglandin F2 alpha given once between d 25 and 40 postpartum (n = 190). No profertility effects were detected in cows, regardless of their health status during the periparturient period, except cows given gonadotropin-releasing hormone between d 11 and 18 had shorter intervals to first estrus and to first service than controls. Cows with reproductive disorders (abnormal health status) in the concurrent lactation had longer intervals from calving to conception after receiving gonadotropin-releasing hormone between d 18 and 25 or prostaglandin F2 alpha between d 33 and 40. Abnormal health status adversely affected every reproductive trait measured. Early postpartum treatments with either hormone failed to improve reproductive performance of dairy cows, in contrast to several reports of profertility effects for gonadotropin-releasing hormone, and a few reports for prostaglandin F2 alpha.     

Early postpartum reproductive profiles in Holstein cows with retained placenta and uterine discharges

Sixty Holstein cows were allocated to three groups. Twenty cows had retained placenta. The remaining cows were examined on d 14 postpartum and those with purulent discharges (n = 22) were assigned to one group and the remaining (n = 18) to a control group. Within each group, cows were given randomly either gonadotropin-releasing hormone (i.m., 200 micrograms) or saline on d 15 postpartum to evaluate the effect on changes in ovarian structures and plasma progesterone through 50 d postpartum and fertility. Corpora lutea were found in control cows by d 21, cows with uterine discharge by d 28, and cows with retained placenta by d 27. Maximum progesterone production during the first luteal phase was higher in control cows than in cows with purulent discharge or retained placenta (4.66 ng/ml compared with 3.23 and 3.34 ng/ml, respectively). Duration of the first corpus luteum was affected by clinical condition. Only 6.2% of cows with retained placenta had corpora lutea of normal duration (12 to 16 d), whereas 43.8 and 50.0% of cows with uterine discharge and control cows had normal postpartum luteal phases. Measures of fertility were not affected by gonadotropin-releasing hormone. Control cows had less days to conception (97) and fewer services per conception (1.6) than cows with retained placenta (134 and 2.5, respectively). Clinical group affected reproduction more than gonadotropin-releasing hormone did, possibly by altering 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.     

Effects of gonadotropin-releasing hormone and human chorionic gonadotropin on pregnancy rate in dairy cattle

This study was to determine if gonadotropin-releasing hormone, human chorionic gonadotropin, or their interaction would affect pregnancy rate or milk progesterone profiles in dairy cattle. Cows and heifers eligible for first, second, and third services were assigned to four treatments: 1) saline i.m. within 5 min after AI (d 0) and saline 15 d after AI (n = 222 AI); 2) gonadotropin-releasing hormone (100 micrograms) i.m. within 5 min after AI and saline on d 15 (n = 223); 3) saline after AI and human chorionic gonadotropin (3500 IU) i.m. on d 15 (n = 196); 4) gonadotropin-releasing hormone at AI and human chorionic gonadotropin on d 15 (n = 195). Progesterone was quantified in milk samples collected twice weekly for 5 wk from all cows. Neither gonadotropin-releasing hormone, human chorionic gonadotropin, gonadotropin-releasing hormone x human chorionic gonadotropin interaction, age, nor service number affected pregnancy rate, which averaged 46.2%. Pregnancy and human chorionic gonadotropin affected milk progesterone profiles. Pregnancy maintained progesterone concentrations and human chorionic gonadotropin on d 15 increased them. This study does not support the general use of gonadotropin-releasing hormone at AI or human chorionic gonadotropin 15 d after AI as methods for enhancing pregnancy rates of dairy cattle.     

Reproductive and metabolic characteristics of dairy cattle supplemented with beta-carotene

Seventy-eight Holstein cows alternately were assigned at calving to receive beta-carotene supplementation or act as controls to determine effects of beta-carotene on reproduction and carotene, luteinizing hormone, progesterone, insulin, glucose, and glucagon concentrations in blood plasma. Cows were fed a corn silage-based complete ration. Biweekly jugular blood samples were collected beginning the week after parturition through 90 days. At day 30, supplemented cows received 600 mg synthetic beta-carotene daily for 60 days. Plasma carotene reached a peak of 2.45 micrograms/ml compared to 1.50 micrograms/ml in controls. Supplementation significantly increased plasma carotene but had no effect on luteinizing hormone, progesterone, insulin, glucose, glucagon, or reproductive measures. Days to first heat, days to first breeding, days open, and services per conception averaged 74, 74, 95, and 1.7 for supplemented cows and 64, 76, 102, and 1.9 for control cows. Progesterone increased as lactation progressed. Somatic cells were not different between supplemented and control cows. Supplementation of beta-carotene did not improve reproductive efficiency or alter luteinizing hormone, progesterone, insulin, glucose, or glucagon in blood plasma or affect somatic cells in 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.     

Comparison of methods for the synchronization of estrous cycles in dairy cows. 2. Effects of progesterone and parity on conception

Three experiments with 571 dairy cows indicated that significantly more primiparous cows given two prostaglandin F2 alpha injections 14 d apart conceived within 30 d of first AI than did cows given two injections 11 d apart (84 vs. 62%). Fewer multiparous cows given two injections 14 d apart and inseminated after estrus conceived than did cows given two injections and a progesterone intravaginal coil inserted 8 d after the first injection (42 vs. 66%). Fewer cows given one injection of prostaglandin and inseminated at estrus conceived than did cows given two injections 14 d apart and a progesterone coil (39 vs. 66%). Conception rates of cows given two prostaglandin injections were positively related to plasma progesterone concentrations 2 and 4 d before the second injection (r = .81 and .86). Cows with progesterone concentrations below 5 ng/ml, 2 d before the second prostaglandin injection, and synchronized by two prostaglandin injections or by two injections and a progesterone coil had conception rates of 36 and 63%, respectively. Cows with progesterone concentrations above 5 ng/ml had a conception rate of 75 and 46% in the two treatments, respectively. Conception after estrus synchronization depends on the method and on the plasma concentrations of progesterone. Progesterone coils may be used before AI to enhance fertility in cows with low progesterone concentrations.     

Relationship of plasma beta-carotene and vitamin A to luteal function in postpartum cattle

The influence of plasma concentrations of beta-carotene and vitamin A on in vivo progesterone production by bovine corpora lutea after gonadotropin-releasing hormone-induced LH release was assessed in 39 postpartum dairy cows. Thirty Holsteins and nine Jerseys were given 100 micrograms gonadotropin-releasing hormone on d 12 of an estrous cycle, which began from 30 to 49 d postpartum. Concentrations of beta-carotene and vitamin A in plasma and progesterone and LH in serum were determined prior to gonadotropin-releasing hormone injection (0 h); serum progesterone and LH concentrations were also determined 1, 2, and 3 h after injection of gonadotropin-releasing hormone. Serum concentrations of progesterone and LH were increased by gonadotropin-releasing hormone. Incremental progesterone production in an analysis of covariance was influenced by breed as well as the interactions of breed with vitamin A, of season with beta-carotene, and of season with vitamin A. The regression coefficients were positive for beta-carotene and negative for vitamin A in all cases. In conclusion, luteal function in the postpartum cow appears to be related to plasma concentrations of beta-carotene and vitamin A.     

Effect of dietary protein and exogenous gonadotropin-releasing hormone on circulating progesterone concentrations and performance of Holstein cows

Sixty-three Holstein cows were used in a 2 x 2 factorial arrangement of treatments to determine the effects of dietary protein and exogenous gonadotropin-releasing hormone on performance postpartum and on circulating concentrations of progesterone. The experimental diets, 14 or 20% CP (dry basis), were fed from parturition to 14 wk postpartum. Blood serum urea N was elevated in the cows fed the 20% versus 14% CP diet. Feed intake was increased, but yield and composition of milk were not affected by the higher protein diet. Circulating concentrations of progesterone were lower in cows fed 20% than 14% protein on d 12 of both the synchronized and subsequent estrous cycles. Exogenous gonadotropin-releasing hormone administered on d 12 of the preceding estrous cycle did not increase the concentration of progesterone on d 12 of the subsequent estrous cycle. Luteal phase concentrations of progesterone were reduced by high dietary protein, but were not affected by gonadotropin-releasing hormone given on d 12 of the preceding cycle.     

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.     

Milk progesterone in postpartum and pregnant cows as a monitor of reproductive status.

Milk samples were analyzed for progesterone content by a petroleum ehter extraction and competitive protein binding assay validated for milk. In one experiment, 11 cows were sampled twice daily for 24 days beginning with an observed estrus 15 to 45 days postpartum, and again 19, 21, 23, and 25 days after breeding. Progesterone values during the estrous cycle paralleled those for blood plasma but were slightly higher at estrus (1.49 ng/ml milk) and maximum (9 ng/ml) on days 11 to 16 of the estrous cycle. After breeding, cows later diagnosed pregnant averaged 7.12 ng/ml while those later found to be nonpregnant averaged 2.36 ng/ml. All diagnoses of pregnancy were correct. In a separate experiment there was no difference between milk from front and rear quarters, but progesterone was highest in last milk, intermediate in composite milk, and lowest in first milk.