Early Detection of Pregnancy and Embryonic Loss in Dairy Cattle by ELISA Tests

Over a 25-month period 8118 blood samples were assayed for the presence of the serum pregnancy specific-protein B (PSPB) and progesterone (P4) concentrations on three Hungarian large-scale dairy farms. Pregnancy (n = 4085) was checked by BioPRYN assay at 30-36 days post-insemination (PI). Samples from all cows that tested not pregnant and from cows with an optical density (OD) reading in the BioPRYN test that was between 0% and 30% above the cutoff OD value were tested for serum P4 concentration. According to serum P4 concentration, cows were assigned to three categories: high (>4 ng/ml), medium (2-4 ng/ml) and low (<2 ng/ml) serum progesterone. The authors predicted a presumed (low) or possible (medium) late embryonic loss (LEL) or maintenance of the pregnancy (high). A total of 710 LELs were detected (17.4%) and 31.8% of them were predicted because of a low OD value at 30-36 days after insemination. Lower PSPB serum level significantly refers for LEL (p < 0.0001). The prediction rate for the true embryonic loss was 31.8% when OD cutoff from 0% to + 30% of cutoff was examined while it was 62.5% when the threshold was OD cutoff of 0% to 10% of cutoff. The authors conclude that BioPRYN was useful for prediction of a part of LEL in dairy cows and serum P4 concentration in these cows related to the rate of LEL.     

Milk Progesterone Profile at and after Artificial Insemination in Repeat-Breeding Cows: Effects on Conception Rate and Embryonic Death

The aim of this study was to investigate whether the skim milk progesterone concentrations at artificial insemination (AI) and day of rise of post-ovulatory progesterone concentration thereafter affect the conception and embryonic death rates in repeat-breeding cows. Milk samples were obtained from 96 repeat-breeding cows that failed to conceive to three or more AIs. The samples were taken from the cows at the day of AI and three times/week until day 45 post-AI. Skim milk was obtained after centrifugation and used for progesterone assay. The cows with a progesterone concentration more than 0.5 ng/ml at AI showed a significantly higher incidence of late embryonic death than those having a progesterone concentration<0.5 ng/ml at AI (p<0.01). As the progesterone level at insemination rose, conception rate declined. A negative correlation was shown between conception rate and skim milk progesterone level at AI. Of 56 cows showing a rise of progesterone to 1 ng/ml or more within 6 days after AI, 28 cows (50%) conceived. On the contrary, only eight of 39 cows (20.5%) conceived when the progesterone rose up to 1 ng/ml after day 6 post-AI. We concluded that increased progesterone concentration at the time of AI and delayed rise of progesterone post-AI might lead to decrease in fertility in repeat-breeding cows.     

The comparison of treating Holstein dairy cows with progesterone, CIDR and GnRH after insemination on serum progesterone and pregnancy rates

This experiment was conducted to investigate the effects of different exogenous progesterone sources administrated after artificial insemination (AI) on serum progesterone (P(4) ) concentration and pregnancy rates in Holstein lactating cows. Sixty-four lactating Holstein dairy cows were allocated to four different treatments (n = 16 per treatment): the cows 1) were injected with physiological saline on days 5 and 13 after AI (control group); 2) were injected with progesterone on days 5 and 13 after AI (P group); 3) received controlled internal drug releasing device (CIDR) for a period from day 5 to 19 after AI (CIDR group); and 4) were injected with gonadotropin-releasing hormone (GnRH) agonist on days 5 and 13 after AI (GnRH group). Blood samples were collected on days 0 (AI day), 5, 13, 16 and 19 after AI to determine serum P(4) concentration. The results revealed a significant difference among treatment groups for serum P(4) concentration on days 13, 16 and 19 with the lowest concentration of serum P(4) for the control group. The pregnancy rate was also positively affected by all the treatments with CIDR having the greatest effect on pregnancy rate. Overall, the results indicated that CIDR has the greatest effect on serum P(4) concentration and pregnancy rate, although the administration of P and GnRH during days after AI increased serum P(4) concentration in lactating dairy cows as well.     

Relationship of Progesterone,Bovine Pregnancy-Associated Glycoprotein-1 and Nitric Oxide with Late Embryonic and Early Fetal Mortalities in Dairy Cows

The aim of the present study was to determine the relationship of progesterone (P4), bovine pregnancy-associated glycoprotein-1 (bPAG-1) and nitric oxide (NO) levels with late embryonic (LEM; day 28 to day 42) and early fetal mortalities (EFM; > day 42 to day 56) in dairy cows. Transrectal ultrasonography (6–8 MHz) was performed in 100 Holstein-Friesian cows at days 28, 42 and 56 after artificial insemination (AI; day 0) to diagnose pregnancy and to monitor the fate of the embryo. After ultrasound scanning of each cow, a milk sample was collected for assessment of P4 by an ELISA test and a blood sample was collected for assessment of bPAG-1, by using a double-antibody radioimmunoassay, and serum NO metabolites (nitrate + nitrite). Based on ultrasonographic examinations and bPAG-1-RIA, 41 of 100 inseminated cows were confirmed pregnant at day 28 after AI. Nine cows suffered of LEM, and 6 cows suffered of EFM and the overall pregnancy loss rate was 36.6% (15/41) between days 28 and 56 of pregnancy. By logistic regression analysis, there were no significant relationships between the level of P4 and bPAG-1 at day 28 after AI and the occurrence of LEM and EFM. Also, there were no significant relationships between the levels of P4 and bPAG-1 at day 42 and the occurrence of EFM. On the other hand, a significant relationship (P<0.05) was found between NO level at day 28 and the occurrence of LEM. In conclusion, measurement of the serum NO concentration at day 28 of pregnancy might help to predict the outcome of pregnancy by day 42 in dairy cows but further studies are needed to confirm this.     

Serum Progesterone Concentrations and Blood Neutrophil Alkaline Phosphatase Activity in Cows Following Normal and Abnormal Parturition

Serum progesterone concentrations and alkaline phosphatase (ALP) activity of blood neutrophils were determined in 3 groups of cows (n = 5 each) on days 1 and 2 and then at 3-day intervals up to 32 days post-partum. Group I cows had a normal delivery, Group II cows had dexamethasone-induced parturition and Group III cows were subjected to a caesarian section. All cows in Group III and 2 cows in Group II retained their fetal membranes. Mean serum progesterone concentrations declined the second day after calving (to < 0.67 ng/ml) and remained at low levels (< 0.54 ng/ml) throughout the observation period, except for the values in Group III, which were elevated on day 16 (0.94 ng/ml), declined again on day 26 (0.46 ng/ml) and peaked (1.05 ng/ml) on day 32 portpartum. Significant (P < 0.01) differences were found between serum progesterone concentrations on day 1 and on each of the other sampling days in Groups I and III. Day X parturition group interaction was significant (P < 0.05) for the progesterone concentrations. No significant differences were found between the overall means of ALP activity of blood neutrophils in the 3 parturition groups nor between days of the experiment. No significant correlation was found between serum progesterone concentrations and ALP activity values of blood PMN during the first 32 days post-partum. Inhalt: Serum Progesteron Konzentrat und Aktivität der alkalischen Phosphatase in neutrophilen Blutzellen bei Küken mit normalem und abnormalen Geburtsverlauf Serum-Progesteronkonzentrationen und alkalische Phosphatase- (ALP) Aktivität von neutrophilen Granulozysten, nach der Abkalbung am Tag 1 und 2 und dann im Abstand von 3 Tagen bis zum 32. Tag p.p. bei Kühen untersucht, die in drei Untersuchungsgruppen (n = 5) eingeteilt waren. Gruppe I kalbte normal ab, in Gruppe II wurde die Geburt durch Dexamethason eingeleitet, und Kälber der Gruppe III wurden via Kaiserschnitt gewonnen. Alle Tiere der Gruppe III und 2 Kühe der Gruppe II wiesen Nachgeburtsverhaltungen auf Der mittlere Serum-Progesterongehalt sank am 2. Tag p.p. auf < 0,67 ng/ml ab und blieb auf niederigem Niveau (< 0,54 ng/ml) während der gesamten Untersuchungs-periode. Lediglich in Gruppe III, mit erhöhten Werten (0,94 ng/ml) an Tag 16 p.p., sanken die Werte wieder an Tag 26 p.p. (0,46 ng/ml) und erreichten einen Maximalwert an Tag 32 p.p. mit 1,05 ng/ml. Signifikante (P 0,01) Unterschiede in der Serum-Progesteronkonrentration wurden zwischen Tag 1 p.p. und allen anderen Untersuchungstagen in den Gruppen I und III gefunden. Interaktionen zwischen den Gruppen für Tag X waren signifikant (P 0,05). Keine signifikanten Unterschiede wurden für den Gesamtmittelwert der ALP-Aktivität in den neutrophilen Granulorysten zwischen den Gruppen und im Vergleich der Untersuchungstage gefunden. Es wurden auch keine signifikanten Korrelationen zwischen den Serum-Progesteronkonzentrationen und der ALP-Aktivität der PMN während der ersten 32 Tage p.p. nachgewiesen.     

Suppression of ovarian progesterone production in dairy cows using an implant of GnRH-agonist (deslorelin) for the purpose of evaluating progesterone metabolism

OBJECTIVE: To evaluate the potential of an implant of a GnRH-agonist (deslorelin) to create a progesterone free animal suitable for studying progesterone (P4) metabolism in intact cows by measuring blood P4 and faecal P4 metabolites. METHODS: Experiment 1: Eighteen non-lactating cycling Holstein-Friesian cows, 4 to 7 years old, were allocated to one of three groups to study plasma P4 concentrations preceding an intravaginal insert. These groups comprised: i) a deslorelin group (GnRH-agonist implanted); ii) a PGF group receiving two injections of prostaglandin (PGF2alpha) 12 days apart; and, iii) an ovariectomised (OVX) group. An intravaginal device (CIDR) was inserted into the vagina of each animal and left in place for 11 days. Plasma P4 concentrations were measured during the study period. Experiment 2: Twelve non-lactating cycling Holstein-Friesian cows, 4 to 7 years old, were allocated to two groups: i) a deslorelin group (GnRH-agonist implanted); and ii) an ovariectomised group. Plasma P4 and faecal P4 metabolites (20-oxo-pregnanes, 20alpha-OH and 20beta-OH) were monitored for a period of 5 weeks. RESULTS: Experiment 1: Average plasma P4 concentration did not differ between the three groups (1.28, 1.43 and 1.55 ng/mL for deslorelin, OVX and PGF cows, respectively, P = 0.8) during the period of supplementation. Experiment 2: There was no difference in plasma P4 (mean plasma P4 < 0.02 ng/mL, P = 0.9) and faecal P4 metabolites between deslorelin and OVX cows 2 weeks after the implantation (P = 0.7). CONCLUSIONS: These data showed that a GnRH-agonist (deslorelin) implant may be used as an alternative to ovariectomy to create a progesterone free animal suitable for studying the metabolism of administered P4.     

Effect of types of breeding on embryo survival following first AI in lactating Holstein cows

The main objective was to investigate the effects of timed-AI protocols versus AI following oestrus detection on circulating progesterone (P4) and embryo survival after first service in Holstein cows. Cycling status was determined by ultrasonography and by plasma P4 concentrations 14 and 26 days after calving, and only cows with a corpus luteum and/or P4 ≥ 1 ng/ml were used. Cows were randomly allocated to one of three types of breeding: DO (n = 80), received GnRH-7d-PGF2α-3d-GnRH and Ovsynch56 was initiated 7 days later; G7G (n = 70), received PGF2α-2d-GnRH and Ovsynch56 (GnRH-7d-PGF2α-56h-GnRH-16h-AI) was initiated 7 days later; or AI based on oestrus detection, EDAI (n = 60). Progesterone was also determined at AI and 8, 16, 18 and 20 days after AI; ISG15 and MX2 mRNA abundance were determined 16 days after AI. Mean plasma P4 at AI was greater in the EDAI group compared with DO and G7G groups, while after AI, P4 was greater in DO and G7G groups compared with EDAI group. However, the percentage of cows with a concentration of P4 < 0.8 ng/ml at AI did not differ among groups. Relative mRNA abundance of ISG15 and MX2 was greater in the DO and G7G groups compared to those in EDAI group. Pregnancy per AI 16, 32 and 60 days after AI was greater (p < .05) in cows in the DO group compared with those in EDAI group (47.5%, 38.8% and 36.3% vs. 30.0%, 21.7% and 15.0%). Pregnancy losses between 16 and 60 days after AI were greater (p < .05) in cows in the EDAI (50.0%) group compared to those subjected to DO (23.7%) or G7G (24.1%). In conclusion, the use of timed-AI synchronization protocols resulted in greater circulating P4 concentrations post-AI and greater embryo survival following first service in lactating Holstein cows.     

Cortisol and luteinizing hormone after adrenocorticotropic hormone administration to postpartum beef cows

Cortisol and luteinizing hormone (LH) were measured in serum after the administration of adrenocorticotropic hormone (ACTH) to suckled (S) and nonsuckled (NS) beef cows. Blood was sampled on 2 consecutive days every 2 weeks for four bleeding periods starting 14 days after calving. Cows were injected with 200 IU ACTH or saline in a 2-day switchback design. Serum was collected before ACTH or saline injection and at 30-min intervals thereafter for 8 hours. Average cortisol concentrations in serum were similar in S and NS cows (6.4 +/- .6 and 6.1 +/- .8 ng/ml, respectively) after saline. Average cortisol concentrations in serum collected during an 8-hr period after ACTH on days 14, 28, 42 and 56 postpartum were 24.7 +/- 2.4, 31.8 +/- 3.5, 36.4 +/- 4.2 and 40.7 +/- .5 ng/ml, respectively, for S cows, and 31.1 +/- 2.9, 44.7 +/- 5.2, 45.0 +/- 5.7 and 46.0 +/- 5.4 ng/ml, respectively, for NS cows. Cortisol response to ACTH, measured as area under the response curve, was greater (P less than .05) in NS than in S cows. Amount of cortisol released by 200 IU ACTH was maximal by days 28 to 29 postpartum in NS cows, but the response increased gradually between days 14 to 15 and days 56 to 57 in S cows. overall, LH in serum averaged .55 +/- .08 ng/ml for S cows and .92 +/- .06 ng/ml for NS cows after saline, and .49 +/- .07 ng/ml for S cows and .94 +/- .06 ng/ml for NS cows after ACth. Although mean and peak serum LH concentrations did not differ between cows given ACTH and those given saline, the number of LH peaks and the number of cows having LH after saline. Mean serum LH concentrations were lower (P less than. 05) in S than in NS cows at 28 days postpartum. The number of LH peaks was lower (P less than .05) and the magnitude of the largest LH peak tended to be lower (P less than .06) in S cows at all sampling periods.     

Effect of naloxone on serum luteinizing hormone, cortisol and prolactin concentrations in anestrous beef cows

Two experiments were conducted with the opioid antagonist naloxone to determine the effect of opioid receptor blockade on hormone secretion in postpartum beef cows. In Exp. 1, nine anestrous postpartum beef cows were used to measure the effect of naloxone on serum luteinizing hormone (LH), cortisol and prolactin concentrations. Cows received either saline (n = 4) or 200 mg naloxone in saline (n = 5) iv. Blood samples were collected at 15-min intervals for 2 h before and after naloxone administration. Serum LH concentrations increased (P less than .01) in naloxone-treated cows from 1.8 +/- .04 ng/ml before treatment to 3.9 +/- .7 ng/ml and 4.2 +/- .5 ng/ml at 15 and 30 min, respectively, after naloxone administration. In contrast, LH remained unchanged in saline-treated cows (1.6 +/- .3 ng/ml). Serum cortisol and prolactin concentrations were not different between groups. In Exp. 2, 12 anestrous postpartum beef cows were used to examine the influence of days postpartum on the serum LH response to naloxone. Four cows each at 14 +/- 1.2, 28 +/- .3 and 42 +/- 1.5 d postpartum received 200 mg of naloxone in saline iv. Blood samples were taken as in the previous experiment. A second dose of naloxone was administered 2 h after the first, and blood samples were collected for a further 2 h. Serum LH concentrations increased (P less than .01) only in cows at 42 d postpartum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Applicability of a progesterone-based timed artificilal insemination protocol after follicular fluid aspiration using the ovum pick-up technique in suckled beef cows

We conducted a progesterone-based timed AI protocol after follicular fluid aspiration using the ovum pick-up (OPU) technique to examine its applicability to the suckled beef cow. A total of 19 beef cows were randomly allocated to one of the following three groups based on the number of days postpartum: 13 to 60 days (Group A: suckled; early postpartum period, n=9), 61 to 150 days (Group B: suckled; mid postpartum period, n=6), or 151 to 281 days (Group C: non-suckled; prolonged open period, n=4) postpartum. These cows were treated with follicular fluid aspiration and insertion of a progesterone-releasing intravaginal device (PRID) on day 0. The PRID was removed and 500 microg of cloprostenol was intramuscularly administered on day 7. A dose (100 microg) of fertirelin acetate was injected intramuscularly 48 hours later, and this was followed by a timed AI (TAI) after another 18 hours (day 10). Serum samples were taken on days 0, 7, 9, 10, 12, 17, 24 and 31 for determination of the estradiol-17beta (E(2)) and progesterone concentrations. Pregnancy diagnosis was made by rectal palpation approximately 60 days after TAI. There was no significant difference in the peripheral E(2) concentrations among the three groups during the period of the hormonal treatment. The average progesterone concentrations in Group A on day 17 were significantly higher than those in Group B and exceeded 1.0 ng/ml on day 17 and thereafter. There was no significant difference in the numbers of collected immature oocytes among the three groups. The pregnancy rates in Groups A, B, and C were 77.8% (7/9), 83.3% (5/6) and 50.0% (2/4), respectively. In conclusion, this timed AI protocol is applicable to suckled beef cows within the period of 60 days postpartum.     

Evaluation of a fixed-time artificial insemination protocol for postpartum suckled beef cows

Treatment with melengestrol acetate (MGA), an oral progestin, prior to administration of gonadotropin-releasing hormone (GnRH) and prostaglandin F2alpha (PG) effectively synchronizes estrus and maintains high fertility in postpartum beef cows. The objective of this experiment was to determine whether treatment with MGA prior to a GnRH-PG-GnRH protocol would improve pregnancy rates resulting from fixed-time artificial insemination (AI). Multiparous crossbred beef cows at two University of Missouri-Columbia farms (n = 90 and n = 137) were assigned by age and days postpartum to one of two treatments. Cows were fed carrier (1.8 kg x animal(-1) x d(-1)) with or without MGA (0.278 mg x kg(-1)) for 14 d. All cows were administered GnRH (100 microg; intramuscularly) on d 12 after MGA or carrier withdrawal and 7 d before PG (25 mg; intramuscularly). All cows received a second injection of GnRH and AI 72 h after PG. Mean days postpartum for MGA and control cows at the initiation of treatment were 39.6 and 38.9 d for herd 1; and 51.9 and 50.9 d for herd 2, respectively (P > 0.70 within herds). Blood samples were collected from all cows at 10 and 1 d before the feeding of MGA or carrier began and at the times GnRH and PG were administered. Concentrations of progesterone in serum at the initiation of treatment were elevated (>1 ng/mL) in 0% of MGA and 7% of control cows in herd 1, and 54% of MGA and 49% of control cows in herd 2 (P > 0.05 within herds). Pregnancy rates to fixed-time AI were determined by transrectal ultrasonography 50 d after AI. Pregnancy rates in herd 1 were 58% (26/45) and 51% (23/45) for MGA-treated and control cows, respectively (P = 0.52), and 63% (44/70) and 45% (30/67) for MGA-treated and control cows in herd 2, respectively (P = 0.03). Differences in pregnancy rates to fixed-time AI were significant (P = 0.04) when data from the two herds were combined (with MGA = 70/115 [61%]; control = 53/112 [47%]). There was no difference (P > 0.20) in final pregnancy rates (timed AI plus 45 d exposure to bulls) between treatments, within herds, or when herds were combined. In summary, pregnancy rates resulting from fixed-time AI may be improved with treatment of MGA prior to a GnRH-PG-GnRH protocol.     

Temporal pattern of insulin-like growth factor-I response to exogenous bovine somatotropin in lactating cows

The effect of exogenous bovine somatotropin (bST) treatment on the temporal pattern of insulin-like growth factor-I (IGF-I) in serum of four multiparous Holstein cows was examined. Cows (190 +/- 24 days postpartum) were treated with daily subcutaneous injections of recombinant bST (40 mg) or excipient for 12-day periods in a crossover experimental design. During excipient treatment, concentrations of IGF-I in serum were relatively constant throughout the day and averaged 70 ng/ml. Following the first bST injection, serum IGF-I began increasing after a lag of 5 to 7 hr and progressively increased over the first 2 days of treatment. Serum IGF-I levels were approximately 2-fold greater than control values at the end of day 1 of bST treatment, with a 3-fold elevation observed at the end of day 2. Concentrations of IGF-I in serum plateaued by day 3 of bST treatment. Serum concentrations of IGF-I did not follow the oscillating pattern of bST in serum resulting from daily bST injections. Milk yield (3.5% fat-corrected) plateaued after 6 days of bST treatment and was increased 61% (+15.3 kg). Both IGF-I and milk yield remained essentially constant across days for the remainder of treatment. Following cessation of treatment, serum IGF-I and milk yield gradually declined, returning to control values after approximately 4 days. The temporal pattern of circulating concentrations of IGF-I is consistent with a role for IGF-I in mediating a portion of the effects of exogenous bST in lactating cows.     

Relationship between oxidative stress and the success of artificial insemination in dairy cows in a pasture-based system

This study was designed to evaluate whether the outcome of artificial insemination (AI) was affected by the metabolic and oxidative status of dairy cows. Seventy-nine inseminations in 40 cows, were classified, on the basis of blood progesterone (P4) and pregnancy-associated glycoprotein (PAG) concentrations and clinical confirmation of pregnancy into, three categories: (1) positive (AI+, resulted in pregnancy, n=26; 33%), (2) negative (AI-, did not result in pregnancy, n=49; 62%), and (3) embryonic mortality (EM, n=4; 5%). Reactive oxygen metabolites, biological antioxidant potential, oxidative stress index, body condition score, glucose, total proteins, albumin, urea, non-esterified fatty acids (NEFAs), cholesterol, triglycerides, haptoglobin and advanced oxidative protein products (AOPPs) were measured on the day of AI (day 0), and 30 and 42days later. Cows with EM had lower BCS scores (2.5) than AI+ (2.8) and AI- (2.9) cows (P<0.05). During the post-partum period, body condition score (BCS) increased and NEFAs decreased (P<0.05) suggesting a recovery from the negative energy balance (NEB). The only significant differences found were that the mean concentration of AOPPs was higher and that of albumin lower in EM cows than in AI+ and AI- (P<0.05) animals. Plasma concentration of reactive oxygen metabolites and biological antioxidant potential were not related to AI outcome. Further studies are required to confirm this finding and to clarify the role of oxidative status on cows' fertility.     

Evaluation of factors that affect embryonic loss in dairy cattle

OBJECTIVE: To identify potential risk factors for embryonic loss before 35 to 42 days of gestation in dairy cattle. DESIGN: Prospective observational study. ANIMALS: 381 cows. PROCEDURE: Body condition score was determined at the time of artificial insemination (AI; day 0) and on days 20, 23, and 27 and between days 35 and 41; serum progesterone concentration was measured on days 0; 20 or 21; and 23, 24, or 25. Cows were excluded from analyses if day 0 serum progesterone concentration was > or = 1.0 ng/mL and classified as pregnant on day 23 if serum progesterone was > 1.5 ng/mL on day 20 or 21 and day 23, 24, or 25. Cows were examined via transrectal ultrasonography on day 27 or 28 and rectally palpated for pregnancy on days 35 to 41. RESULTS: 39% of cows that were pregnant on day 23 lost their embryo by day 27, and 18% of cows that were pregnant on day 27 or 28 were not pregnant on days 35 to 41. Breeding a pregnant cow posed the greatest risk for embryonic loss at both time periods. Mean serum progesterone concentrations on day 21 or 22 and day 23, 24, or 25 were lowest for cows that lost an embryo between days 24 and 28. Cows with a linear somatic cell count score > 4.5 before AI were twice as likely to lose the embryo by 35 to 41 days, compared with cows with a score < 4.5. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that embryonic loss could be reduced by more accurate estrus detection, reducing mastitis, and strategies to improve progesterone concentration after breeding.     

Is nutritional anestrus precipitated by subfunctional corpora lutea in beef cows?

Thirty-four multiparous, lactating, cyclic beef cows which calved in moderate body condition were used to determine effects of restricted nutrition on corpus luteum (CL) development and endocrine status. At 78 d postpartum, six cows were assigned to a control (CON) diet (26.0 Mcal ME), fed to increase bodyweight (BW) and body condition score (BCS), and the remaining 28 cows were fed to lose BW and BCS on a restricted (RES) diet (14.0 Mcal ME). Following a 40-d adjustment period on respective diets, estrous cycles were synchronized and cows bled daily for determination of progesterone (P4), luteinizing hormone (LH) and insulin (INS) beginning at the synchronized estrus. Ultrasonography was used to determine the ovulatory follicle and CL development. Control cows were maintained for one estrous cycle and were ovariectomized on day 11 of their second cycle. Ten cows on restricted diet (RES-C) continued to form a functional CL (P4 > 1.5 ng/ml at day 10 of an estrous cycle) through as many as 5 cycles, after which observations were discontinued. Fourteen cows on restricted diet (RES-A) were ovariectomized on day 11 of a cycle when a CL was identified by ultrasonography, but was subfunctional (P4 < 1.5 ng/ml on day 10 of that cycle). Four additional RES-A cows which had subfunctional CL were not ovariectomized but were bled for an additional 25 d. At ovariectomy, CL and ovarian weights were collected. Luteal tissue was prepared for evaluation of P4 synthesis, LH responsiveness in vitro, and for determination of P4 content and total LH receptors. Bodyweight and BCS increased in CON cows; whereas, RES cows lost BW and BCS (P < .05). In the cycle prior to ovariectomy, serum P4 and LH were not different in 18 RES-A cows which developed subfunctional CL in comparison to CON cows. Four RES-A cows not ovariectomized but bled for an additional 25 d neither exhibited estrus, ovulated, nor had P4 concentrations greater than .3 ng/ml. Serum INS was lower in RES-A cows during the cycle prior to ovariectomy than in CON cows (P < .05). During the 11-d period prior to ovariectomy, mean serum P4 and INS were lower in RES-A cows than in CON cows (P < .05); however, serum LH was not different. Furthermore, CL and ovarian weights, P4 content of CL, secretion of P4 by luteal tissue in response to LH in vitro and LH receptor number were not different between CON and RES-A cows. In conclusion, nutritional anestrus may be preceded by the formation of a CL with lower steroidogenic output in vivo. However, luteal tissue, collected from RES-A cows, did not appear to be subfunctional during in vitro incubation when substrate availability and gonadotropin support were equal between diets.     

Milk progesterone determination in buffaloes post-insemination

Milk progesterone levels were studied in 17 buffaloes between post-insemination and 30 days thereafter. Six animals were confirmed pregnant. Seven of 10 animals took greater than 24 days to return to oestrus and the pattern of change of progesterone levels indicated a steady increase from 1 ng/ml on day 2 to 20 ng/ml on day 14 post-insemination, falling subsequently to about 5 ng/ml, 2 days prior to oestrus. In pregnant buffaloes, the milk progesterone levels showed a continuous increase up to 20 days post-insemination but did not decline thereafter. In individual buffaloes returning to oestrus, a cut-off milk progesterone level of greater than 10 ng/ml was considered for pregnancy diagnosis, 20, 22 and 24 days post-insemination. The test was 60, 75 and 75% accurate for detection of pregnancy on days 20, 22 and 24 respectively and 100% for non-pregnancy diagnosis on all three days. Individual animals showed a highly variable oestrous cycle length, which has been suggested as a contributory factor for false pregnancy diagnosis by milk progesterone test in this species. Milk progesterone assay may also identify silent heats, and clinical disorders.     

Role of the sensitivity of detection of oestrus in the submission rate of cows treated to resynchronise oestrus

OBJECTIVE: To determine if failure to detect oestrus in cows treated to resynchronise oestrus leads to fewer cows being inseminated than are truly in oestrus. PROCEDURE: Cows in three herds were enrolled in a controlled breeding program that involved synchronisation of oestrus for a first round of artificial insemination (AI) followed by resynchronisation for a second round of AI. Just before oestrus was expected at the second round of AI, aids for the detection of oestrus were fitted, which included pedometers, radiotelemetric transmitters (HeatWatch), tail-paint and heatmount detectors. Milk samples were collected at the second round of AI (day 33, herds A and B; day 35, herd C of the treatment program) and were used in combination with pregnancy testing to determine the number of cows that were in oestrus (milk progesterone < 2.0 ng/mL) and cows that were not in oestrus (milk progesterone > 2.0 ng/mL or pregnant at second round of AI) at the time samples were collected. RESULTS: The mean sensitivity of detection of oestrus at the resynchronised oestrus was 92.5% and did not differ significantly between herds (P = 0.19). A total of 75% (60/80) of cows that were retrospectively determined to be not pregnant at the time of the second round of AI were classified as having high (> or = 2.0 ng/mL) concentrations of progesterone in milk at that time. Pregnancy testing of cows about 35 days after AI suggested that early pregnancy loss also contributed to a reduction in submission rates at the resynchronised oestrus. CONCLUSION: Failure to submit cows for insemination at a resynchronised oestrus was mainly due to cows not being in oestrus rather than due to a failure to detect oestrus.     

Serum Profiles of Pregnancy-Associated Glycoprotein, Oestrone Sulphate and Progesterone During Gestation and Some Factors Influencing the Profiles in Ethiopian Borana and Crossbred Cattle

This study presents serum concentrations of pregnancy‐associated glycoprotein (PAG), oestrone sulphate (E1‐S) and progesterone (P4), and the effects of some dam and foetus‐related factors on these profiles during gestation in Borana and crossbred cattle. The PAG concentrations at 4th week post‐conception ranged from 1.5–5.5 and 2.1–4.7 ng/ml in Borana (n = 6) and crossbred (n = 8) cattle, respectively. The mean PAG concentrations increased progressively from 4th to 33rd week of gestation (from 3.3–173 ng/ml for Borana and 4.2–240 ng/ml for crossbred cattle) and reached peak around calving. Breed, parity status, dam body weight, foetal sex and foetal birth weight significantly influenced the PAG concentrations. After delivery, the PAG concentrations declined steadily to 5.7 ng/ml in Borana (n = 7) and 3.9 ng/ml in crossbred (n = 6) cattle 10 weeks post‐partum. The serum E1‐S concentrations at 17th week of pregnancy ranged from 0.3–2.6 and 0.9–5.7 ng/ml in Borana (n = 8) and crossbred (n = 9) cattle, respectively. The mean E1‐S concentrations increased progressively from 17th to 33rd week of gestation (from 1.1–4.6 ng/ml for Borana and 2.7–10.8 ng/ml for crossbred). Breed, parity status, dam body weight and foetal sex significantly influenced E1‐S concentrations. The P4 concentrations at 4th week of pregnancy ranged from 3.2–5.1 and 1.7–8.9 ng/ml in Borana (n = 6) and crossbred (n = 8) cattle, respectively. The P4 level remained elevated throughout pregnancy. This study indicated that the serum PAG and P4 concentrations at 4th and E1‐S approximately 17th week of pregnancy were above the cut‐off value for pregnancy test and the hormonal profiles observed were comparable to the previous reports. Furthermore, the PAG and E1‐S profiles were considerably influenced by factors such as breed, weight and parity status of the dam, and foetal sex and foetal birth weight (only PAG).     

奶牛发情期的奶孕酮分析

<正> 影响奶牛人工授精成功率的主要原因是对发情期能否正确判断,适时输精。据1981年联邦德国Bavarian动物健康服务机构的报告,输精时间不正确,是影响奶牛繁殖率的重要因素。由于孕酮含量在发情周期中呈规律性变化,在发情期显著下降,因此可     

Effect of morphine on serum gonadotropin concentrations in postpartum beef cows

Morphine (M), an opioid agonist, was administered to postpartum (PP) Angus cows to investigate opioid modulation of gonadotropin secretion. In Exp. 1, eight PP cows (36.9 +/- 2.3 d) received either M (1 mg/kg; n = 4) or saline solution (S) (n = 4) via i.v. injection 36 h after calf removal. Morphine decreased (P less than .01) the number of serum LH pulses (3.0 +/- 1.1 pre- vs .3 +/- .3 post-pulses/h) and, compared with pretreatment values (3.3 mg/ml), decreased (P less than .05) mean LH at 105 min (2.1 ng/ml) through 270 min 1.9 ng/ml +/- .4). Serum prolactin (PRL) increased (P less than .01) following M from 16.4 ng/ml to a peak of 59.3 ng/ml (+/- 3.9). Serum FSH concentrations were unaffected. In Exp. 2, M (.31 mg/kg i.v. injection followed by .15 mg/(kg.h) infusion; n = 6) or S (n = 6) treatments were given for 7 h beginning 36 h after calf removal. Serum LH was similar between groups during the pretreatment and the first 6 h of infusion, but M decreased (P less than .001) the number of serum LH pulses (.44 +/- .09 vs .06 +/- .04 pulses/h). Morphine increased (P less than .05) serum PRL. It is concluded that M differentially modulated gonadotropin secretion in the cow such that PRL increased, LH decreased and FSH was unchanged.