Fetal survival in the cow after pregnancy diagnosis by palpation per rectum

It is desirable to determine wheter a cow has failed to become pregnant as early as possible, preferably prior to 50 d after insemination. Although palpation per rectum has been the generally accepted method of pregnancy diagnosis in cattle, the procedure may be a significant iatrogenic cause of fetal attrition. In a study conducted at a North Florida dairy from January through June 1982, pregnancy was determined in 192 Holstein-Friesian cows by measuring low milk progesterone (P4) content on day of insemination (Day 0) and elevated P4 on Days 21 and 24. Pregnant cows were randomly assigned to a treatment and a control group. Cows in the treatment group (n = 85) were palpated per rectum twice between Days 42 and 46 after insemination. Cows in the control group (n = 107) were not palpated until both groups were palpated at Day 90. Palpation, done by two experienced clinicians, consisted of palpation of fetal fluid fluctuation, identification of the amniotic vesicle, and slipping of the chorioallantoic membranes. In both groups of cows fetal viability was monitored by milk P4 content. Last milk (5 to 15 ml) was collected from one front quarter on Days 0, 21, and 24 and twice weekly thereafter through Day 63. Milk was defatted by centrifugation and the fat-free milk progesterone content measured by a radioimmunoassay without extraction. The milk P4 test was 80.0% accurate in determining pregnancy in the palpated and nonpalpated cows. In the cows palpated on Days 42 to 46, pregnancy rates declined by 7.5% as determined by palpation at Day 90, or by 11.4% as determined by milk P4 content through Day 63 (both values P < 0.05). Cows that were not palpated on Days 42 to 46 showed a 1.9% increase or a 4.3% decline in pregnancy rates as determined by the same criteria. Before palpation, at Days 42 to 46, pregnancy rates were better in cows that were inseminated in winter (January to March) than in spring (April to June) (82.3% vs 61.6%; P < 0.05); P4 content was higher (winter > spring = 2.13 ng/ml vs 1.38 ng/ml; P < 0.05). First-lactation cows had higher P4 values on Days 21 and 24 than older cows (P < 0.01).     

Progesterone concentrations in milk fat at first insemination--effects on non-return and repeat-breeding

The relationship between milk fat progesterone concentration at first artificial insemination (AI) and reproductive performance of Norwegian Red Cattle dairy cows was investigated in a field study. Fifteen AI technicians collected milk samples from 2250 cows from 458 herds and progesterone was analysed in the milk fat portion of the samples. Logistic regression revealed decreased probability of non-return, and increased likelihood of repeat-breeding, with increasing progesterone concentrations at insemination. The odds ratio for non-return and for repeat-breeding between the minimum and maximum value for milk fat concentrations between 4 and 32.2ng/ml was 4.5 (P < 0.01) and 3.2 (P = 0.01), respectively. The variable 'Technician' did not significantly influence non-return rate or repeat-breeding. Progesterone concentration at AI was positively related to calving to last insemination interval, and to number of inseminations per cow (P < 0.05). The association between the progesterone concentration at insemination and calving interval was only marginally significant. Technician was significantly (P < 0.01) associated with interval from calving to first AI, interval from calving to last AI (P = 0.05), and number of AI per cow (P = 0.01). Technician was not significantly related to the calving interval.We conclude that increased suprabasal progesterone concentrations at the time of first insemination, and higher return rate at AI may, at least partly, be due to endocrinological asynchrony at AI, a condition which may lead to decreased fertility.     

Conception rates and serum progesterone concentration in dairy cattle administered gonadotropin releasing hormone 5 days after artificial insemination

The objectives of this study were to determine the effect of administration of exogenous GnRH 5days after artificial insemination (AI) on ovarian structures, serum progesterone concentration, and conception rates in lactating dairy cows. In experiment 1, 23 Holstein cows were synchronized using the Ovsynch protocol. Five days after AI (day 0) cows were assigned randomly to receive either saline (saline; n=11) or 100microg GnRH (GnRH; n=12). To examine ovarian structures, ultrasonography was performed on day 1 and every other day beginning on day 5 until day 13. On days 5 and 13 blood samples were obtained to measure serum progesterone concentrations. All cows in the GnRH-treated group developed an accessory corpus luteum (CL), whereas cows in the saline group did not. Mean serum progesterone concentrations did not differ between GnRH and saline groups on day 5 (1.64+/-0.46ng/ml versus 2.04+/-0.48ng/ml). On day 13 serum progesterone concentrations were greater (P<0.05) in the GnRH group compared with saline (5.22+/-0.46ng/ml versus 3.36+/-0.48ng/ml). In experiment 2, 542 lactating cows, at two different commercial dairies, were used to test the effect of administering GnRH 5 days after AI on conception rates. Cows were synchronized and detected for estrus according to tail chalk removal. Cows detected in estrus received AI within 1h after detection of estrus. Five days after AI, cows were assigned randomly to receive either GnRH (n=266) or saline (n=276). Pregnancy status was determined by palpation per rectum of uterine contents approximately 40 days after AI. There was no effect of farm on conception rate. There was no effect of treatment as conception rates did not differ between GnRH and saline groups (26.7% GnRH versus 24.3% saline). Regardless of treatment, days in milk, parity, milk yield, and number of services had no effect on the odds ratio of pregnancy. In summary, the results of this study indicated that GnRH administered 5 days after AI increased serum progesterone by developing an accessory CL but did not improve conception rates in dairy cattle.     

Association between leptin single nucleotide polymorphism and reproductive performance of lactating Holstein cows

The objectives of the current study were to evaluate the associations among reproductive performance of Holstein cows, single nucleotide polymorphism (SNP) in the R4C locus of the Exon-2 region of the leptin gene and health disorders. Lactating dairy cows (n=814) had their DNA sequenced at the Exon-2 region of the leptin gene to determine the presence of SNP in the R4C locus. Cows had the stage of estrous cycle synchronized with two injections of PGF(2α) 14d apart, with or without a progesterone insert. After the second PGF(2α), cows were either inseminated on detected estrus and, if not inseminated within 13d, cows were submitted to a timed artificial insemination (AI) protocol 13d after the second PGF(2α). Blood was sampled at 35±3, 49±3, and 62±3d in milk (DIM) and cows with progesterone <1.0ng/mL in the first two samples were considered to be anovular, whereas those with at least one of the first two samples with progesterone ≥1.0ng/mL were considered to have initiated estrous cyclicity. Anovular cows with progesterone ≥1.0ng/mL at 62±3DIM were considered to have resumed estrous cyclicity. Pregnancy was diagnosed at 31 and 60d after first postpartum AI and at 42d after subsequent inseminations. Resulting genotypes were CC (34.6%), CT (48.2%), and TT (17.2%). Leptin genotype was associated (P=0.03) with increased prevalence of estrous cyclic cows at 49±3DIM, as TT cows were and tended to be less likely to be estrous cyclic than CC and CT cows, respectively. Other measures of reproductive performance in the first 305DIM were not associated with leptin genotype. Cows diagnosed with at least one postparturient disease were less likely to become pregnant after first (P<0.01) and second (P=0.02) AI and to be pregnant at 305DIM (P<0.01). Furthermore, cows diagnosed with a disease event had a 36% reduction (P<0.001) in the rate of pregnancy, which extended the median interval to pregnancy by 60d. Estrous cyclic cows had a 35% increased rate of pregnancy, which reduced the interval to pregnancy by 34d. Cows homozygous for the T allele were less likely to be cyclic early postpartum, but leptin genotype was not associated with other reproductive responses when cows were subjected to a controlled breeding program.     

Genetic and environmental factors influencing first service conception rate and late embryonic/foetal mortality in low fertility dairy herds

The objective of this study was to identify factors affecting variation in conception rate to first artificial inseminations (AI) (CR: number of pregnant cows on D80-100/inseminated cows) and the incidence of embryonic/foetal loss (LEM) between 21 and 80 days of pregnancy (number of cows non-pregnant on D80-100/pregnant on D21) in 44 low fertility dairy herds of the west-central region of France. Reproductive status was assessed using progesterone milk concentration on D0 = Day of AI and D21-24, plasma PSPB concentration on D30-35, rectal palpation on D80-100 and observed return to oestrous. The final data set contained 1285 Prim'Holstein cows, 5.0% (64/1285) were inseminated in the luteal phase (progesterone > or = 3 ng/ml on D0), 61.3% (787/1285) were pregnant on D21-24 (progesterone < 3 ng/ml on D0 and > or = 5 ng/ml on D21-24), 15.4% lost their embryo/foetus between D21-24 and D80-100 (198/1285) and 45.8% (589/1285) were pregnant on D80-100. The incidence of late embryonic/foetal loss (LEM) was 25.2% (198/787). Multivariate logistic regression models including the random herd effect were used to analyse the relationship between AI centre, AI sire, cow's sire, parity, interval between calving and AI, milk production, milk protein content, body condition score (BCS) on D0, season of calving, season of AI, estimated genetic index on CR and LEM incidence. CR was significantly related to parity (p < 0.05), milk production after calving (p < 0.05) and estimated genetic value (p < 0.01). A significant difference in CR was observed for calving to AI interval > or = 70 days versus > or = 90 days, but the overall effect of the interval was not significant (p = 0.11). LEM incidence was affected by period of AI (p < 0.05), milk production (p < 0.05) and BCS (p < 0.05), but was not related to estimated genetic index. In conclusion, in these low fertility herds, the incidence of LEM was high and 25% of the cows lost their embryo after 21 days of pregnancy. LEM was affected by specific factors (season, BCS), which were not related to CR. The absence of a relationship between estimated genetic index and LEM in spite of its effect on CR indicates that estimated genetic merit has a greater effect on early embryonic loss or fertilisation failure than on later stages of embryo development.     

Progesterone concentration, follicular development and induction of cyclicity in dairy cows receiving intravaginal progesterone inserts

Objectives were to evaluate progesterone concentrations after cows had initiated estrous cycles following calving and induction of estrous cycles in postpartum anovular high-producing Holstein dairy cows treated with controlled internal drug releasing (CIDR). In experiment 1 (EXP1), 62 cows that had initiated estrous cycles received a new CIDR (NCIDR) containing 1.38 g of progesterone or a 7-d used autoclaved CIDR (UCIDR) 48h after luteolysis for 7 d. Ovaries were examined by ultrasonography, and plasma analyzed for concentrations of progesterone. In experiment 2 (EXP2), 515 cows diagnosed as anestrus were randomly assigned to untreated control, NCIDR or UCIDR for 6d. Plasma was analyzed for concentration of progesterone 12 d after CIDR removal to determine ovulation. In EXP1, milk yield and body condition did not influence progesterone concentrations. Concentration of progesterone tended to increase faster (P=0.10) in cows receiving UCIDR than NCIDR, but both treatments reached a plateau at 90min. Cows receiving the NCIDR had greater (P=0.04) concentrations of progesterone during the 7-d treatment, but they were mostly subluteal (<1.0 ng/mL) after d 2. After removal, concentrations of progesterone were greater for NCIDR than UCIDR for the first 45 min, and were similar thereafter. Multiparous cows had lesser (P=0.004) concentrations than primiparous cows throughout the study. The pattern of ovarian follicular development was not affected by treatment. In EXP2, induction of onset of estrous cycles increased (P<0.01) with progesterone treatments, but was similar between NCIDR and UCIDR. Proportion of cows experiencing shorter than typical length estrous cycles after first AI tended to be greater (P=0.09) for control cows than those receiving the CIDR, and for cows remaining anestrous than those in which onset of estrous cycles was induced. Pregnancy per AI and pregnancy loss were similar among treatments. Cows that resumed estrous cyclicity prior to first AI had greater (P=0.01) pregnancy per AI. Treatment of high-producing Holstein cows that had previously initiated onset of estrous cycles with CIDR resulted in subluteal concentrations of progesterone, but in anestrous high-producing cows increased induction of estrous cycles with no effect on fertility at first insemination.     

Fixed-time artificial insemination of postpartum beef cows at 72 or 80 h after treatment with the MGA Select protocol

The objective was to determine the appropriate timing of fixed-time artificial insemination (AI) following administration of the MGA Select protocol. Cows at two locations (Location 1, n=114; Location 2, n=97 ) were assigned to fixed-time AI at 72 or 80 h by age, body condition score (BCS), days postpartum (DPP), AI technician, and sire. All cows were synchronized with the MGA Select protocol, consisting of oral administration of melengestrol acetate (MGA; 0.5mg/hd per day) for 14 days, GnRH (Cysotrelin, 100 microg, i.m.; Day 26) 12 days after MGA withdrawal, followed in 7 days with PGF(2alpha) (PG; Lutalyse, 25mg i.m.; Day 33). Cows were inseminated at 72 h ( n=108 ) or 80 h ( n=103 ) after PG and GnRH (100 microg) was given at insemination. Location was not significant and, therefore, was removed from the model. Mean BCS ( 5.2+/-0.1, 72 h; 5.3+/-0.1, 80 h) and DPP ( 34+/-2, 72 h; 35+/-2, 80 h) did not differ ( P>0.1 ) between treatments. Serum progesterone concentrations 7 and 1 day prior to MGA were used to determine pre-treatment cyclicity: cows with at least one sample with progesterone > or =1 ng/ml were defined as cyclic (33/108, 31%, 72 h, versus 32/103, 31%, 80 h; P>0.1). Cows with serum progesterone concentrations > or =1 ng/ml on the day of PG were defined as responding to the synchronization protocol (74/108 (69%), 72 h versus 69/103 (67%), 80 h; P>0.1 ). Although pregnancy rates were higher ( P<0.05 ) for cows inseminated at 72 h (69/108, 64%) versus 80 h (52/103, 50%) after PG, pregnancy rates at the end of the breeding season did not differ ( P>0.1 ) between treatments (98/108 (91%), 72 h; 88/103 (85%), 80 h). In conclusion, pregnancy rates were higher when postpartum beef cows synchronized with the MGA Select protocol were inseminated at 72 h versus 80 h after PG.     

Timed artificial insemination plus heat II: gonadorelin injection in cows with low estrus expression scores increased pregnancy in progesterone/estradiol-based protocol

The use of tail chalk and estrus/heat expression scores (HEATSC) evaluation is instrumental in identifying cows with greater estrus expression and greater artificial insemination pregnancy rates (P/AI) in cows submitted to timed artificial insemination (TAI), and cows with low or no estrus expression present lower P/AI. It was intended in this study to improve the pregnancy rates in TAI for Bos indicus beef cows, and gonadotrophin-releasing hormone (GnRH) injection was hypothesized to increase pregnancy rates in a TAI program for cows submitted to progesterone–estradiol-based protocols with low or no estrus expression, evaluated by HEATSC. Cows (n= 2284) received a progesterone device and 2 mg estradiol benzoate, after 8 days the device was removed and 1 mg estradiol cypionate, 150 μg of d-cloprostenol and 300 IU equine chorionic gonadotropin was administered. All cows were marked with chalk and HEATSC evaluated (scales 1 to 3) at TAI performed on day 10. Animals with HEATSC1 and HEATSC2 (n= 937) received 100 μg de gonadorelin (GNRH group; n= 470), or 1 ml saline (Control group; n= 467), and cows with HEATSC3 (named HEAT group; n= 1347) received no additional treatment. The larger dominant follicle, evaluated on day 8and at TAI (day 10), was greater in HEAT group (P= 0.0145 and P <0.001, respectively). Corpus luteum (CL) area and progesterone concentration was evaluated on day 17, and CL area was larger in HEAT group, intermediary in Control and lower in GnRH group (Control= 2.68 cm², GnRH= 2.37 cm², HEAT group= 3.07 cm², P <0.001). Greater progesterone concentrations were found in HEAT group than in Control and GnRH groups (Control= 4.74 ng/ml, GnRH= 4.29 ng/ml, HEAT group= 6.08 ng/ml, P<0.001). There was a difference in ovulation rate, greater in HEAT group than GnRH and Control groups (Control= 72.5%; GnRH= 81.25%; HEAT group= 90.71%; P= 0.0024). Artificial insemination pregnancy rates was greater in HEAT group (57.09% (769/1347) than in Control and GNRH groups, with positive effect of GnRH injection at the time of TAI in P/AI (Control= 36.18% (169/467), GnRH= 45.95% (216/470); P<0.0001). In conclusion, GnRH application in cows with low HEATSC (1 and 2) is a simple strategy, requiring no changes in TAI management to increase pregnancy rates in postpartum beef cows submitted to progesterone–estradiol-based TAI protocols, without reaching, however, the pregnancy rates of cows that demonstrate high estrus expression at the TAI.     

Use of cow-side progesterone tests to improve reproductive performance of high-producing dairy cows

This study evaluated the effectiveness of the cow-side ELISA milk progesterone test in improving postpartum reproductive performance in the Dordt College dairy herd. Cows that produced more than 18,500 lb of milk per lactation were assigned to the high production group (40 cows), while cows that produced less than 18,500 lb of milk (42 cows) were assigned to the low production group. Twenty-one cows in the high production group and 19 cows in the low production group received no ELISA testing (untreated controls), while the remaining cows in each group were evaluated by ELISA test every 7 d beginning on Day 27 post partum (treated cows). A sequence of 2 high progesterone tests and 1 low test indicated the cows were cycling normally. Cows that had low milk progesterone levels (<5 ng/ml) for 3 consecutive tests were assumed to have follicular cysts and were treated with 2 ml GnRH (Cystorelin, 50 mug/ml). Cows that had 3 consecutive high tests (>5 ng/ml) were assumed to have persistent corpora lutea (CL) and were treated with 5 ml PGF(2)alpha (Lutalyse, 5 mg/ml). In both the high and low production groups, treated cows had higher (P < 0.08) pregnancy rates by Day 210 than the untreated controls (63.2 vs 38.1% and 56.5 vs 42.1%, respectively). The days open were reduced (P < 0.05) for the treated animals by 41.6 d compared with the controls. The treated cows produced a net savings of $70.42 (US) per cow assuming a $3.00 savings/day open.     

Effect of repeated injections of GnRH on reproductive parameters in postpartum anestrous dairy cows

To induce cyclicity in dairy cattle with prolonged postpartum anestrous, repeated dosages of gonadotrophin releasing hormone (GnRH) were administered. Twenty-one (21) Holstein dairy cows and heifers calving between October 1, 1989, and January 1, 1990, at the Louisiana State University Dairy were used in the study. The animals were defined as anestrous if their plasma progesterone remained < 1.0 ng/ml until 32 to 36 days post partum. They were randomly assigned to one of two treatment groups. Group 1 (n=6) received two injections 1 hour apart of a GnRH analogue (50 mug) (i.m.). The treatment was repeated twice weekly at 3- to 4-day intervals. Group 2 controls (n=6) received saline (1 ml, i.m.) on the same schedule as Group 1. A maximum of 12 to 13 treatments were given. Cattle that had plasma progesterone >1.0 ng/ml by 32 to 36 days post partum were identified as Group 3, or cyclic contemporaries (n=9). Postpartum anestrous in the herd was 46.2% (18 39 ). Cows in Group 1 had significantly fewer days to first plasma progesterone > 1.0 ng/ml than those in Group 2 (P < 0.05), but more days than Group 3. Cows in Group 1 also had significantly fewer treatments to induce plasma progesterone > 1.0 ng/ml than those in Group 2 (P < 0.05). There were no significant differences among treatment groups in the number of days from calving to first observed estrus or the number of days open (P > 0.05).     

Timed artificial insemination plus heat I: effect of estrus expression scores on pregnancy of cows subjected to progesterone–estradiol-based protocols

Expression of estrus near timed artificial insemination (TAI) is associated with greater fertility, and estrus detection could improve TAI fertility or direct TAI management, although accurate estrus detection can be difficult and time-consuming using traditional methods. The aim of this study is to evaluate influence of estrus on pregnancy (artificial insemination pregnancy rates (P/AI)) and to validate an alternative method to classify estrus/heat expression using tail chalking (HEATSC) in postpartum Bos indicus cows subjected to TAI in progesterone–estrogen-based protocols. In experiment 1 (Exp. 1), cows (5491) were subjected to visual observation of estrus after progesterone device removal, before TAI, and P/AI was evaluated according to estrus and body condition score (BCS). Cows received a progesterone device and 2 mg estradiol benzoate (EB). After 8 days, the device was removed and 150 μg of d-cloprostenol and 300 IU equine chorionic gonadotrophin was given. Later, animals in Exp. 1 received 1 mg EB and TAI 44 to 48 h. In the Exp. 2 – 3830 cows using similar protocol, received different ovulation inducers: 1 mg EB (n=1624) or 1 mg estradiol cypionate (EC; n=2206) on day 8 (D8). Cows were then marked with chalk, and HEATSC evaluated at TAI on D10 (HEATSC1 – no chalk removal=no estrus expression; HEATSC2 – partial chalk removal=low estrus expression; HEATSC3 – near complete/complete chalk removal=high estrus expression). In Exp. 1, cows showing estrus presented greater P/AI (48.4% v. 40.2%, P<0.05). In Exp. 2, P/AI (HEATSC1 – 40.0%; HEATSC2 – 49.7%; HEATSC3 – 60.9%; P<0.001), and larger follicle timed artificial insemination (LFTAI) (<0.001) varied according to HEATSC. There was no difference in P/AI (P=0.41) or LFTAI (P=0.33) according to ovulation inducer. Cows with greater BCS showed greater P/AI in both experiments (P<0.05). Estrus presence and greater HEATSC improved P/AI, and EC v. EB used promoted differential estrus manifestation (cows showing HEATSC2 and HEATSC3: 79.5% with EB v. 69.98% with EC use, P<0.001), however, with similar P/AI. The use of HEATSC in B. indicus cows subjected to TAI is useful to identify cows with greater estrus expression and consequently improved pregnancy rates in TAI, allowing the cows with low HEATSC to be targeted for additional treatments aimed at improving P/AI.     

Suppression of luteal function in dogs by luteinizing hormone antiserum and by bromocriptine

Beagle bitches were treated with equine anti-LH serum (ALHS) or the dopamine agonist bromocriptine at selected times during the 2-month luteal phase of the ovarian cycle or pregnancy. After a single injection of ALHS (10 ml, i.m.) at Day 42 of pregnancy (N = 2) or the ovarian cycle (N = 3), progesterone was reduced (P less than 0.05) to 7-24% of preinjection values within 1-2 days, and by 4-8 days returned to levels not different from those in control bitches treated with normal horse serum. Injections of bromocriptine (0.1 mg/kg, i.m.) daily for 6 days caused abrupt declines in progesterone which lasted 6-8 days in bitches treated at Day 8 or 22 of pregnancy (N = 5). In bitches treated at Day 42 of pregnancy (N = 3) or in non-pregnant cycles (N = 4) the bromocriptine treatment caused declines (P less than 0.05) in progesterone which were permanent, extensive (less than 2 ng/ml), and therefore abortive. The declines in progesterone in response to immunoneutralization of LH and to prolactin-lowering doses of a dopamine agonist demonstrate that normal luteal function in dogs requires both LH and prolactin.     

The effects of ovarian function on estrus synchronization with PGF in dairy cows

Milk progesterone concentration (P4), milk yield, milk composition, ovarian structures and pregnancy status were studied in 108 cows treated with two doses of PGF 14 days apart and inseminated at fixed time (TAI) 80-82 h later. The synchronization protocol was started at 70+/-1.4 days after parturition. Milk P4 profiles revealed that anestrus, failure of luteolysis following treatment with PGF and failure to ovulate following luteolysis were the main reasons for low pregnancy rate with TAI. Anestrous cows had a higher percentage of milk fat (P<0.05) and higher fat to protein ratio (P<0.01), and cows that did not undergo luteolysis had higher milk yield (P<0.05) and lower percentage of milk protein (P<0.05) than cows that responded to PGF treatment. Cows that did not undergo luteolysis and cows that did not ovulate following luteolysis had lower milk P4 during the luteal phase preceding the second PGF injection (P<0.01 and P<0.05, respectively). Pregnancy rates 24 and 47 days after TAI in cows that responded as expected to the synchronization treatment were 62% and 54%, respectively. Pregnancy was precluded in non-responsive cows. The largest follicle at the time of TAI in cows experiencing late embryonic mortality was smaller (P=0.02) than in cows that successfully maintained pregnancy. Results suggest that a primary reason for low pregnancy rate in dairy cows after administration of PGF and TAI is inappropriate ovarian function prior to, or following treatment.     

Comparison of pregnancy rates with two estrus synchronization protocols in Italian Mediterranean Buffalo cows

The aim in this study was to compare two estrus synchronization protocols in buffaloes. Animals were divided into two groups: Group A (n=111) received 100 microg GnRH on Day 0, 375 microg PGF(2alpha) on Day 7 and 100 microg GnRH on Day 9 (Ovsynch); Group B (n=117) received an intravaginal drug release device (PRID) containing 1.55 g progesterone and a capsule with 10mg estradiol benzoate for 10 days and were treated with a luteolytic dose of PGF(2alpha) and 1000 IU PMSG at the time of PRID withdrawal. Animals were inseminated twice 18 and 42 h after the second injection of GnRH (Group A) and 60 and 84 h after PGF(2alpha) and PMSG injections (Group B). Progesterone (P(4)) concentrations in milk samples collected 12 and 2 days before treatments were used to determine cyclic and non-cyclic buffaloes, and milk P(4) concentrations 10 days after Artificial insemination (AI) were used as an index of a functional corpus luteum. Cows were palpated per rectum at 40 and 90 days after AI to determine pregnancies. All previously non-cyclic animals in Group B had elevated P(4) (>120 pg/ml milk whey) on Day 10 after AI. Accordingly, a greater (P<0.01) relative percentage of animals with elevated P(4) 10 days after AI were observed in Group B (93.2%) than in Group A (81.1%). However, there was no difference in overall pregnancy rates between the two estrus synchronization protocols (Group A, 36.0%; Group B 28.2%). When only animals with elevated P(4) on Day 10 after AI were considered, pregnancy rate was higher (P<0.05) for animals in Group A (44.4%) than Group B (30.3%). The findings indicated that treatment with PRID can induce ovulation in non-cyclic buffalo cows. However, synchronization of estrus with Ovsynch resulted in a higher pregnancy rate compared with synchronization with PRID, particularly in cyclic buffalo.     

Use of milk progesterone enzyme immunoassay for early pregnancy diagnosis in cows

An enzyme immunoassay using beta-galactosidase as the tracer was applied to determine milk progesterone level in cows. The novel method was reliable and practicable and required only a spectrophotometer and a centrifuge as major equipment. The milk progesterone enzyme immunoassay successfully diagnosed early pregnancy in cows. Milk samples were collected from 268 Holstein-Friesian cows in commercial dairy herds on 20, 21 or 22 days(usually 21 days) after insemination. Progesterone level in skim milk higher than 1.0 ng/ml indi-cated pregnancy. Pregnancy was confirmed by rectal palpation on 60 to 120 days after insemination. The accuracy of the milk progesterone test was 60.0 % (132 220 ) for a positive diagnosis and 100 % (48 48 ) for a negative result. A high incidence of embryonic death, 27.9 % (51 183 ), may have reduced accuracy for a positive test result. The enzyme immunoassay may be an alternative to radioimmunoassay in milk progesterone analysis for pregnancy diagnosis.     

Comparison of artificial insemination versus embryo transfer in lactating dairy cows

Conception rates (CR) are low in dairy cows and previous research suggests that this could be due to impaired early embryonic development. Therefore, we hypothesized that CR could be improved by embryo transfer (ET) compared with AI. During 365 days, 550 potential breedings were used from 243 lactating Holstein cows (average milk production, 35 kg/day). Cows had their ovulation synchronized (GnRH-7d-PGF(2alpha)-3d-GnRH) and they were randomly assigned for AI immediately after the second GnRH injection (Day 0) or for transfer of one embryo 7 days later. Circulating progesterone concentrations and follicular and luteal size were determined on Days 0 and 7. Pregnancy diagnosis was performed on Days 25 or 32 and pregnant cows were reevaluated on Days 60-66. Single-ovulating cows with synchronized ovarian status had similar CR on Days 25-32 with ET (n = 176; 40.3%) and AI (n = 160; 35.6%). Pregnancy loss between Days 25-32 and 60-66 also did not differ (P = 0.38) between ET (26.2%) and AI (18.6%). When single (n = 334) and multiple (n = 57) ovulators were compared, independent of treatment, multiple ovulators had greater (P < 0.001) circulating progesterone concentrations on Day 7 (2.7 ng/ml versus 1.9 ng/ml) and there was a tendency (P = 0.10) for a greater CR in multiple ovulators (50.9% versus 38.1%). However, there was no difference in CR between AI and ET cows with multiple ovulations (50.0% versus 51.7%). In single-ovulating cows, CR tended to be lower for AI than ET in cows ovulating smaller follicles (diameter < or = 15 mm; 23.7% versus 42.3%; P = 0.06) but not average-diameter follicles (16-19 mm; 41.2% versus 37.3%; P = 0.81) or larger (> or =20 mm; 34.3 versus 51.0%; P = 0.36) follicles. Thus, although ET did not improve overall CR in lactating cows, follicle diameter and number of ovulating follicles may determine success with these procedures.     

Synchronization of estrus in yearling beef heifers with the melengestrol acetate/prostaglandin F(2alpha) system: efficiency of timed insemination 72 hours after prostaglandin treatment

Two experiments were conducted to determine the conception rates of heifers time-inseminated following melengestrol acetate/prostaglandin F(2alpha) (MGA/PG) estrous synchronization treatment. In Experiment 1, timed insemination of heifers at 72 h after the PG injection, without regard for behavioral estrus, tended to improve (P < 0.15) the percentage of heifers pregnant to artificial insemination (AI) compared with that of synchronized heifers bred 12 h after they were first detected in estrus. In the timed-insemination treatment, heifers exhibiting behavioral estrus 48 to 72 h after PG tended to have an increased (P < 0.15) conception rate to AI compared with heifers exhibiting estrus within 48 h of PG administration. In Experiment 2, the number of heifers conceiving to AI following the MGA/PG estrous synchronization regimen was increased by mass insemination of all heifers not exhibiting estrus by 72 h after PG. The pregnancy rate to AI was higher in heifers with serum progesterone (P(4)) concentrations higher than 1 ng/ml compared with that of heifers with concentrations lower than 1 ng/ml. Of heifers with serum P(4) greater than 1 ng/ml, the pregnancy rate to AI tended to be higher when concentrations exceeded 2 ng/ml than when concentrations were 1 to 2 ng/ml. In cyclic heifers, timed insemination can increase the percentage of heifers pregnant after being synchronized with MGA/PG.     

The effect of strain of Holstein-Friesian cow and feeding system on postpartum ovarian function, animal production and conception rate to first service

Three strains of Holstein-Friesian (HF): high production North American (HP), high durability North American (HD) and New Zealand (NZ) cows were assigned, within strain, to one of three pasture-based feeding systems: (1) the Moorepark (control) system (MP), (2) a high concentrate system (HC), (3) a high stocking rate system (HS). Ovarian function was assessed using milk progesterone samples, collected from 117 cows in each of two successive years, with 81 animals being common to both years. Milk samples were collected thrice weekly, beginning day 5 post-calving and continued to day 26 after first AI. Data from animals subsequent to reproductive hormonal treatment were removed from the analysis. Feed system and strain of HF by feeding system interaction had no significant effect on re-establishment of ovarian activity and subsequent conception rate to first AI. Strain of HF had no significant effect on interval to commencement of luteal activity (CLA). The mean interval to CLA was 32.9 days (S.E. 1.18), ranged from 6 to 100 days, with 42 and 85% of cows ovulating by day 26 and 60, respectively. The HD (62%) and NZ (57%) strains had a higher conception rate to first AI than the HP strain (40%), (P < 0.05). Retrospective analysis categorised all cows into four quartiles based on interval to CLA (< 20 days, 20-26 days, 27-44 days and > 44 days). Cows in the first and fourth CLA quartiles had a longer calving to conception interval (P < 0.05). Cows with abnormal progesterone profiles (38.4%) had an earlier mean calving date, with a similar submission rate and conception rate to first service compared to cows with normal hormonal profiles. There was no significant difference in luteal activity or reproduction performance, apart from calving to conception interval, between cows that conceived or did not conceive to first service. These results indicate that while conception rate to first service differed between strains of HF cow, this was not associated with differences in the onset and pattern of luteal activity post partum.     

Comparison of ovarian palpation, milk progesterone and plasma progesterone in the cow

Holstein cows were examined twice weekly, beginning at 13 to 18 days after parturition, using palpation per rectum and milk progesterone (P4) assay to determine the functional status of the corpus luteum (CL). These results were then compared with P4 concentrations in the plasma to determine which test was the more accurate in detecting functional luteal tissue, or if both tests were needed for optimal accuracy. The tests were found to be comparable except when the plasma P4 concentration was <1. 0 ng/ml. At this level, errors due to palpation occurred more frequently than those due to milk P4 concentrations since the still palpable CL of pregnancy could be mistaken for a functional CL at 14 to 21 days after parturition. However, at all the other concentrations including when plasma P4 was <1.0 ng/ml and the cows were more than 21 days post partum, neither the milk P4 assay nor palpation per rectum could be considered the better indicator of luteal status. Therefore, we conclude that the combined use of both tests does not afford better assessment of luteal function than either test alone.     

Extent and pattern of pregnancy losses and progesterone levels during gestation in Swedish Red and Swedish Holstein dairy cows

Background

Pregnancy loss is a major source of infertility in dairy cows. Despite a fertilization rate after insemination (AI) of approximately 90%, calving rates are 30%–50%, indicating the occurrence of extensive embryonic and foetal losses. The aim of this study was to establish the extent and pattern of embryonic and foetal loss in Swedish Red (SR) and Swedish Holstein (SH) dairy cows, as well as, the relationship to oestrus intensity (OI) and progesterone (P4) concentration. In total, 2130 AIs and 16,176 milk P4 samples from 359 SR and 212 SH dairy cows were included in the study. Pregnancy losses were estimated using data from P4 values combined with AI information and calving data.

Results

Total pregnancy loss from AI to the day of calving was 65%. Early embryonic loss, late embryonic loss and foetal loss were estimated to be 29, 14 and 13%, respectively. There is strong evidence in the literature that P4 concentrations at different time points are associated with pregnancy loss. In the present study, cows with pregnancy losses had significantly higher P4 levels at the day of AI and significantly lower P4 concentration at days 10, 21 and 30 after AI compared to pregnant cows. Swedish Red cows had significantly lower total pregnancy losses compared to SH cows (62% and 68% respectively, P?=?0.017). Early embryonic loss was 6.7% points lower for cows inseminated at a stronger OI (OI?=?3) compared to at a weaker OI (OI?=?2, P?=?0.006). Cows inseminated at ovulation number?≥?5 had significantly lower early pregnancy losses compared to cows inseminated at first or second ovulation (11.5 and 8% points, respectively, P?<?0.05). With an increase of one SD of milk (448 kg ECM) during the first 60 days in milk, early embryonic loss increased by 4.7% points (P?=?0.006).

Conclusions

It is important to increase the number of cows calving per insemination by reducing embryo/foetal loss. This outcome can be achieved by management and breeding for optimal P4 levels at critical time points, and by considering oestrus expression in the breeding programmes to facilitate the correct timing of insemination.