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.     

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.     

Effects of short-term oilseed supplementation on plasma fatty acid composition,progesterone and prostaglandin F metabolite in lactating beef cows

Twenty-four 3-year-old Angus cows (512.2±21.6 kg) and six ruminally cannulated beef heifers (523.1±16.9 kg) were used to determine the impact of feeding oilseeds starting at the beginning of estrous synchronization until maternal recognition of pregnancy on plasma fatty acid composition. Starting ~60 days postpartum cows were synchronized with the Select Synch+controlled internal drug-release (CIDR) device and timed artificial insemination (AI) protocol. The day CIDR was inserted; cattle were randomly assigned to one of the three treatments being grazing only (CON) or a supplement containing whole soybeans (SOY); or whole flaxseed (FLX). Cattle continued to receive these diets for 28 days. Blood was collected every 3 days until 10 days after insemination and then every day until 18 days after insemination. All cattle grazed a common pasture and supplemented cattle were individually fed their respective supplements once daily. Ruminally cannulated heifers were used to evaluate the impact supplements had on forage intake, which was reduced (P=0.05) with oilseed supplementation. Feeding oilseeds increased total fatty acid intake (P<0.001) across treatments with SOY having greater (P<0.001) 18:2n-6 intake than either CON or FLX. Likewise, cattle fed FLX had greater (P<0.001) 18:3n-3 intake than either CON or SOY. There was a treatment×time interaction (P⩽0.05) for all fatty acids identified except for 20:5n-3 (P=0.99). Within 3 days after the start of supplementation, plasma concentrations of 18:2n-6 increased (P<0.001) for cattle fed SOY compared with CON or FLX, whereas flax-fed cattle did not exhibit an increase (P=0.02) until day 15 of supplementation over that of CON. Plasma concentrations for 18:3n-3 was greater (P<0.013) for FLX than both CON and SOY by day 12. Feeding flaxseed tended to (P=0.07) increase and increased (P=0.01) plasma concentrations of 20:4n-6 by day 18 over CON and SOY, respectively. Overall, treatment did not affect serum concentration of progesterone (P=0.18) or prostaglandin F metabolite (P=0.89). However, day after breeding had an effect on serum progesterone (P=0.01) with day 16 after timed AI being lower compared with other days. Feeding oilseeds during the time of estrous synchronization will not only increase the energy density of the diet but will provide key fatty acids around the time of maternal recognition of pregnancy.     

Modifications of the G6G timed-AI protocol improved pregnancy per AI and reduced pregnancy loss in lactating dairy cows

In dairy cows, subjected to a G6G protocol, objectives were to determine effects of (1) extending the interval from prostaglandin F2α (PGF2α) to gonadotropin-releasing hormone (GnRH) during presynchronization; and (2) adding a second PGF2α treatment before artificial insemination (AI), on ovarian response, plasma progesterone (P4) concentrations and pregnancy per AI (P/AI). In a 2×2 factorial design, lactating cows were randomly assigned to one of four timed AI (TAI) protocols: (1) G6G (n=149), one injection of PGF2α, GnRH 2 days later and a 7-day Ovsynch (GnRH, 7 days, PGF2α, 56 h, GnRH, 16 h, TAI) was initiated 6 days later; (2) G6GP (n=144), an additional PGF2α treatment (24 h after the first) during Ovsynch of the G6G protocol; (3) MG6G, one injection of PGF2α, GnRH 4 days later before initiation of the G6G protocol; and (4) MG6GP, an additional PGF2α treatment (24 h after the first) during Ovsynch of the MG6G protocol. Blood samples were collected (subset of 200 cows) at first GnRH and PGF2α of the Ovsynch, and at TAI to measure P4. Ultrasound examinations were performed in a subset of 406 cows to evaluate ovarian response at various times of Ovsynch, and in all cattle to determine pregnancy status at 32 and 60 days after TAI. Extending the interval by 2 days between PGF2α and GnRH during presynchronization increased (P<0.01) ovulatory response to first GnRH of Ovsynch, circulating P4 during Ovsynch, and P/AI at 32 and 60 days after TAI. Adding a second PGF2α treatment before AI increased the proportion of cows with luteal regression (P=0.04), improved P/AI at 60 days after TAI (P=0.05), and reduced pregnancy loss between 30 and 60 days after TAI (P=0.04). In summary, extending the interval from PGF2α to GnRH during presynchronization increased response to first GnRH of Ovsynch and P4 concentrations during Ovsynch, whereas adding a second PGF2α treatment before AI enhanced luteal regression. Both modifications of the G6G protocol improved fertility in lactating dairy cows.     

Feeding dried distillers grains with solubles to lactating beef cows: impact of excess protein and fat on cow performance,milk production and pre-weaning progeny growth

Multiparous Angus×Simmental cows (n=54, 5.22±2.51 years) with male progeny were fed one of two diets supplemented with either dried distillers grains with solubles (DDGS) or soybean meal (CON), from calving until day 129 postpartum (PP) to determine effects of excess protein and fat on cow performance, milk composition and calf growth. Diets were formulated to be isocaloric and consisted of rye hay and DDGS (19.4% CP; 8.76% fat), or corn silage, rye hay and soybean meal (11.7% CP; 2.06% fat). Cow–calf pairs were allotted by cow and calf age, BW and breed. Cow BW and body condition score (BCS; P⩾0.13) were similar throughout the experiment. A weigh-suckle-weigh was performed on day 64 and day 110±10 PP to determine milk production. Milk was collected on day 68 and day 116±10 PP for analysis of milk components. Milk production was unaffected (P⩾0.75) by dietary treatments. Milk urea nitrogen was increased at both time points in DDGS compared with CON cows (P<0.01). Protein was decreased (P=0.01) and fat was increased (P=0.01) in milk from DDGS compared with CON cows on day 68 PP. Compared to CON, DDGS decreased medium chain FA (P<0.01) and increased long chain FA (P<0.01) at both time points. Saturated FA content of milk was decreased (P<0.01) at both time-points in DDGS compared with CON cows, which resulted in an increase (P<0.01) in monounsaturated and polyunsaturated FA, including cis-9, trans-11 conjugated linoleic acid. Daily gain of the DDGS calves was increased (P=0.01) compared with CON calves, resulting in heavier BW on day 129 (P=0.01). Heavier BW of DDGS calves was maintained through weaning (P=0.01). Timed-artificial insemination (TAI) rates were greater for cows fed DDGS compared with cows fed CON (P<0.02), but dietary treatment had no effect on overall pregnancy rates (P=0.64). In summary, feeding DDGS to lactating beef cows did not change cow BW or BCS, but did improve TAI rates and altered milk composition compared with CON. As a result, male progeny from cows fed DDGS during lactation had greater average daily gain and were heavier at day 129 and at weaning compared with male progeny from cows fed a control diet.     

Effect of luteinizing hormone (LH), pregnancy-specific protein B (PSPB), or arachidonic acid (AA) on secretion of progesterone and prostaglandins (PG) E (PGE; PGE(1) and PGE(2)) and F(2alpha) (PGF(2alpha)) by ovine corpora lutea of the estrous cycle or pregnancy in vitro

LH regulates luteal progesterone secretion during the estrous cycle in ewes and cows. However, PGE, not LH, stimulated ovine luteal progesterone secretion in vitro at day 90 of pregnancy and at day 200 in cows. The hypophysis is not obligatory after day 50 nor the ovaries after day 55 to maintain pregnancy in ewes. LH has been reported to regulate ovine placental PGE secretion up to day 50 of pregnancy and by pregnancy-specific protein B (PSPB) after day 50 of pregnancy. The objective of this experiment was to determine if and when a switch from LH to PGE occurred as the luteotropin regulating luteal progesterone secretion during pregnancy in ewes. Ovine luteal tissue slices of the estrous cycle (days 8, 11, 13, and 15) or pregnancy (days 8, 11, 13, 15, 20, 30, 40, 50, 60, and 90) were incubated in vitro with vehicle, LH, AA (precursor to PGE(2) and PGF(2alpha) synthesis), or PSPB in M199 for 4 h and 8 h. Concentrations of progesterone in jugular venous plasma of bred ewes increased (P< or =0.05) after day 50 and continued to increase through day 90. Secretion of progesterone by luteal tissue of non-bred ewes on days 8, 11, 13 and 15 and by bred ewes on days 8, 11, 13, 15, 20, 30, 40, and 50 was increased (P< or =0.05) by LH, but not by luteal tissue from pregnant ewes after day 50 (P> or =0.05). LH-stimulated progesterone secretion by luteal tissue from day 15 bred ewes was greater (P< or =0.05) than day 15 luteal tissue from non-bred ewes. Concentrations of progesterone in media were increased (P< or =0.05) when luteal tissue from pregnant ewes on day 50, 60, or 90 were incubated with AA or PSPB. Concentrations of PGE in media of non-bred ewes on days 8, 11, 13, or 15 and bred ewes on days 8 and 11 did not differ (P> or =0.05). Concentrations of PGE were increased (P< or =0.05) in media by luteal slices from bred ewes on days 13, 15, 20, 30, 40, 50, 60, and 90 of vehicle, LH, AA or PSPB-treated ewes. In addition, PSPB increased (P< or =0.05) PGE in media by luteal slices from pregnant ewes only on days 40, 50, 60, and 90. Concentrations of PGF(2alpha) were increased in media (P<0.05) of vehicle, AA, LH, or PSPB-treated luteal tissue from non-bred ewes and bred ewes on day 15 and by luteal tissue from bred ewes on days 20 and 30 after which concentrations of PGF(2alpha) in media declined (P< or =0.05) and did not differ (P> or =0.05) from non-bred or bred ewes on days 8, 11, or 13. It is concluded that LH regulates luteal progesterone secretion during the estrous cycle of non-bred ewes and up to day 50 of pregnancy, while only PGE regulates luteal progresterone secretion by ovine corpora lutea from days 50 to 90 of pregnancy. In addition, PSPB appears to regulate luteal secretion of progesterone from days 50 to 90 of pregnancy through stimulation of PGE secretion by ovine luteal tissue.     

Saturated fat supplemented in the form of triglycerides decreased digestibility and reduced performance of dairy cows as compared to calcium salt of fatty acids

The two most popular rumen-protected fatty acid supplements in dairy cow rations are calcium salts of palm oil fatty acid calcium salts of palm oil fatty acid (CSFA) and prilled saturated fatty acids (SFAs). The objectives of this study were to determine the effects of supplementing SFA in the form of triglycerides (TSFA), as compared to CSFA, on yields, efficiency and diet digestibility in high-yielding dairy cows. Twenty-eight (14 cows in each group) multiparous cows were fed a basal diet supplemented (on DM basis) with either 12 g/kg TSFA (~350 g/cow per day – contained 980 g/kg fat; 882.3 g/kg SFAs) or 14 g/kg CSFA (~440 g/cow per day – contained 800 g/kg fat; 566.4 g/kg SFAs). The supplement amounts in the diet were balanced according to fat content. Rumen samples were taken for measurements of ammonia and volatile fatty acids concentrations, and fecal samples were taken for digestibility measurements. The CSFA cows produced 3% higher milk yields (47.6 v. 46.2 kg/day; P < 0.0001) and 4.7% higher 4% fat-corrected milk (FCM; 44.7 v. 42.7 kg/day; P = 0.02) than the TSFA cows. No difference in milk-fat content was observed, but milk-protein content was higher in the TSFA than CSFA cows. Yields of fat and protein were similar, but lactose yields were higher in TSFA cows. There were no differences in dry matter intake or efficiency calculations between groups. The ruminal ammonia concentrations were similar between groups, whereas acetate concentrations and acetate : propionate ratio were greater for CSFA than TSFA cows. The apparent total-tract digestibility of dry (P < 0.0007) and organic matters (P < 0.0003), fat (P < 0.0001), NDF and ADF (P = 0.02) were lower in the TSFA v. CSFA cows. In conclusion, the CSFA-supplemented cows produced 3% higher milk and 4.7% higher 4% FCM than the TSFA cows. However, TSFA supplementation did not depress milk-protein content. The apparent total-tract digestibility was lower for all dietary components in the TSFA cows, which was probably due to the effects of both degree of saturation and triglyceride form of the TSFA supplement. Considering that diets were balanced according to the fat content of the supplements, the lower yields of milk and FCM observed in the TSFA than CSFA cows were likely due to the lower digestibility of the fat and other nutrients in the TSFA cows, which might have negatively influenced the dietary energy content.     

Estrous expression during a fixed-time artificial insemination protocol enhances development and interferon-tau messenger RNA expression in conceptuses from Bos indicus beef cows

Expression of estrus (EST) near the time of fixed-time artificial insemination (FTAI) increases pregnancy success in beef females. This outcome has been associated with improved pregnancy establishment and maintenance, although research is still warranted to validate this theory. Hence, this experiment compared ovarian, uterine and conceptus factors associated with pregnancy establishment in Bos indicus beef cows according to estrous expression during a FTAI protocol. One hundred lactating multiparous Nelore cows received a 2 mg injection of estradiol benzoate and an intravaginal progesterone (P4) releasing device on day −11, a 12.5 mg injection of prostaglandin F_2α on day −4, P4 device removal in addition to 0.6 mg injection of estradiol cypionate and 300 IU injection of equine chorionic gonadotropin on day −2, and FTAI on day 0. An estrous detection patch was attached to the tailhead of each cow on day −2, and estrous expression was defined as removal of >50% of the rub-off coating from the patch at FTAI. Overall, 39 cows expressed EST, 55 did not express EST (NOEST), and six cows lost their patch and were discarded from the experiment. Ovarian ultrasonography was performed at FTAI, and on days 7 and 15 of the experiment. Blood samples were also collected on days 7 and 15. Only cows without a corpus luteum (CL) on day 0, and with a CL on days 7 and 15 remained in the experiment (EST, n=36; NOEST, n=48). On day 15, cows were randomly selected within each group (EST, n=29; NOEST, n=30) for conceptus collection via transcervical flushing, followed by endometrial biopsy in the uterine horn ipsilateral to the CL. Within cows not assigned to conceptus collection, blood samples were collected for whole blood RNA extraction (day 20) and pregnancy status was verified by transrectal ultrasonography (day 30). Diameter of dominant follicle on day 0 and plasma P4 concentrations on day 7 were greater (P⩽0.02) in EST v. NOEST cows. Conceptus length and messenger RNA (mRNA) expression of prostaglandin E synthase and interferon-tau were greater (P⩽0.04) in EST v. NOEST cows. Moreover, EST cows diagnosed as pregnant on day 30 had greater (P<0.01) blood mRNA expression of myxovirus resistance 2 on day 20 compared with NOEST. In summary, estrous expression near the time of FTAI enhanced pregnancy establishment factors in B. indicus cows, including conceptus development and mRNA expression of interferon-tau.     

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.     

Minimal progesterone concentration required for embryo survival after embryo transfer in lactating Holstein cows

Objectives were to determine progesterone concentration (P4) from days 4 to 28 relative to presumptive estrus necessary for maintenance of pregnancy in lactating Holstein cows. Cows were assigned to the low P4 (LowP4, n = 28) or control (n = 153) treatments. All cows were presynchronized with two injections of PGF_2α (14 d apart) and an ovulation synchronization protocol (11 d later; GnRH on day −10, PGF_2α on day −3; and GnRH on day 0 = presumptive estrus). Cows in the Low P4 treatment received 2 injections of prostaglandin F_2α on days 4 and 5 (day 0 = presumptive estrus) and a new CIDR insert on day 5 that was replaced every 7 d until day 28. Blood was sampled on days −9, −2, 0, 4, 7,14, 21, and 28. Ovaries were examined with ultrasound on days −9, −3, and 7 and cows bearing a corpus luteum ≥20 mm on day 7 received an embryo. On days 0, 4 and 7 P4 did not differ (P ≥ 0.27) but control cows had greater (P < 0.01) P4 on days 14, 21, and 28. Progesterone concentration fold change from day 0 to 7 was not (P = 0.14) affected by treatment, but P4 concentration fold change from day 7 to 14 was (P < 0.01) greater for control cows compared with LowP4 cows. No LowP4 cows became pregnant after embryo transfer, whereas 35.7, 25.5, and 21.4% of control cows were pregnant on day 28, 42, and 63, respectively. Progesterone concentration fold changes from day 0 to 7 (P = 0.03) and from day 7 to 14 (P = 0.05) were associated with pregnancy outcomes on day 63. Among cows that were pregnant on day 63, the minimum P4 concentration fold changes from day 0 to 7 and from day 7 to 14 were 2.71 and 1.48, respectively. Interestingly, cows with P4 concentration <5 ng/mL on day 14 were (P = 0.01) and tended to be (P = 0.07) more likely to lose pregnancy from day 28 to 42 and from day 28 and 63, respectively. Faster rise in P4 concentration during the metestrus and early diestrus are associated with pregnancy establishment following embryo transfer, which suggests that early rise in P4 concentration has an indirect effect on embryo development through modulation of uterine environment and secretion of histotroph. Furthermore, the positive effects of early rise in P4 concentration appear to go beyond the phase of maternal recognition of pregnancy through adhesion and placentation stages.     

Pregnancy rates to timed artificial insemination in dairy cows treated with gonadotropin-releasing hormone or porcine luteinizing hormone

We compared the effects of porcine luteinizing hormone (pLH) versus gonadotropin-releasing hormone (GnRH) on ovulatory response and pregnancy rate after timed artificial insemination (TAI) in 605 lactating dairy cows. Cows (mean ± SEM: 2.4 ± 0.08 lactations, 109.0 ± 2.5 d in milk, and 2.8 ± 0.02 body condition score) at three locations were assigned to receive, in a 2 × 2 factorial design, either 100 μg GnRH or 25 mg pLH im on Day 0, 500 μg cloprostenol (PGF) on Day 7, and GnRH or pLH on Day 9, with TAI 14 to 18 h later. Ultrasonographic examinations were performed in a subset of cows on Days 0, 7, 10, and 11 to determine ovulations, presence of corpus luteum, and follicle diameter and in all cows 32 d after TAI for pregnancy determination. In 35 cows, plasma progesterone concentrations were determined 0, 3, 4, 5, 6, 7, and 12 d after ovulation. The proportion of noncyclic cows and cows with ovarian cysts on Day 0 were 12% and 6%, respectively. Ovulatory response to first treatment was 62% versus 44% for pLH and GnRH and 78% versus 50% for noncyclic and cyclic cows (P < 0.01). Location, ovulatory response to first pLH or GnRH, cyclic status, presence of an ovarian cyst, and preovulatory follicle size did not affect pregnancy rate. Plasma progesterone concentrations after TAI did not differ among treatments. Pregnancy rate to TAI was greater (P < 0.01) in the GnRH/PGF/pLH group (42%) than in the other three groups (28%, 30%, and 26% for GnRH/PGF/GnRH, pLH/PGF/GnRH, and pLH/PGF/pLH, respectively). Although only 3% of cows given pLH in lieu of GnRH on Day 9 lost their embryo versus 7% in those subjected to a conventional TAI using two GnRH treatments, the difference was not statistically significant. In summary, pLH treatment on Day 0 increased ovulatory response but not pregnancy rate. Cows treated with GnRH/PGF/pLH had the highest pregnancy rate to TAI, but progesterone concentrations after TAI were not increased. In addition, preovulatory follicle diameter did not affect pregnancy rate.     

Resynchronization of ovulation and timed insemination in lactating dairy cows III. Administration of GnRH 23 days post AI and ultrasonography for nonpregnancy diagnosis on day 30

The objective was to compare pregnancy rates to resynchronization and timed AI (TAI) protocols in lactating dairy cows that received GnRH at 23 d and were diagnosed not pregnant at 30 d after the pre-enrollment AI. Nonpregnant cows (624) at ultrasonography on day 30 (study day 0) were classified as diestrus (74.8%), metestrus (5.6%) and without a CL (19.5%). Cows in diestrus were assigned either to the GnRH group (PGF2alpha on day 0, GnRH on day 2 and TAI 16 h later, n = 238) or the estradiol cypionate (ECP) group (PGF2alpha on day 0, ECP on day 1, and TAI 36 h later, n = 229). Cows in metestrus were assigned to the Modified Heatsynch Group (GnRH on day 0, PGF(2alpha) on day 7, ECP on day 8 and TAI on day 9, n = 35). Cows without a CL (n = 122) were classified either as proestrus (10.6%), ovarian cysts (7.5%) or anestrus (1.4%), and assigned to factorial treatments (i.e., use of GnRH versus CIDR) to either the GnRH group (GnRH on day 0, PGF2alpha on day 7, GnRH on day 9 and TAI 16 h later, n = 28), the CIDR group (CIDR insert from days 0 to 7, PGF2alpha on day 7, GnRH on day 9 and TAI 16 h later, n = 34), the GnRH + CIDR group (GnRH on day 0, CIDR insert from days 0 to 7, PGF2alpha on day 7, GnRH on day 9 and TAI 16h later, n = 32), and the control group (PGF2alpha on day 7, GnRH on day 9 and TAI 16 h later, n = 28). For cows without a CL, plasma P4 concentrations were determined on days 0, 7, 10 and 17 and ovarian structures determined on days 0, 7 and 17. Pregnancy rates were evaluated at 30, 55 and 90 d after the resynchronized AI. For cows in diestrus, there were no differences in pregnancy rates on days 30, 55 and 90 for cows in the GnRH (27.5, 26.5 and 24.2%) or ECP (29.1, 25.5 and 24.1%) groups. In addition, there were no differences in pregnancy losses between days 30 and 55 and 55 and 90 between the GnRH (7.0 and 8.6%) and ECP (9.8 and 5.4%) groups. For cows without a CL, GnRH on day 0 increased the proportion of cows with a CL on days 7 and 17 and plasma P4 concentration on day 17 in cows with ovarian cysts but not for cows in proestrus. The CIDR insert increased pregnancy rate in cows with ovarian cysts but reduced pregnancy rate for cows in proestrus.     

Reproductive responses following postpartum suppression of ovarian follicular development with a deslorelin implant during summer heat stress in lactating dairy cows

The objective was to evaluate pregnancy rate to a timed artificial insemination (TAI) protocol in the autumn for cows treated with a non-degradable GnRH agonist implant (Deslorelin [DESL], 5mg) during the summer heat stress period compared with non-treated controls (CON). Cows were randomly assigned to receive or not a DESL implant within 1-4 days postpartum (dpp) twice weekly, from 25 June through 8 August 2001. All cows in DESL implant and CON treatments were injected with PGF(2alpha) 7 days after enrollment. Ultrasonography (US) monitored numbers of ovarian follicles and corpus luteum (CL) at approximately 10, 30, 35/36, 45/44, 56/55 and 66/63dpp, while DESL implants were in situ and concurrently CON, respectively. DESL implants were removed at two specific days, 28 August and 4 September. Cows had DESL implant in situ for a range of 28-67 days, depending on date of enrollment and implant removal. Within 61-100dpp, 31 days after implant removal, DESL implant and CON cows were initiated in a Presynch-Ovsynch and TAI protocol. Pregnancy was evaluated by US and palpation per rectum at 28 and 46 days after TAI, respectively. Plasma concentrations of progesterone were analyzed for sets of blood samples collected during the Presynch-Ovsynch and at TAI day followed 8 days later. Cows in the DESL-implant treatment had more (P<0.01) Class 1 (3-5mm) follicles, less (P<0.01) Class 2 (6-9mm), Class 3 (> or =10mm) follicles and CL compared with CON cows. Proportion of cows having initiated estrous cycles after calving was less (P<0.01) in the DESL-implant treatment (52.2%, 58/111) compared with CON (93.7%, 104/111) at the beginning of Ovsynch. Pregnancy rate to TAI was less (P<0.01) in the DESL implant (27.5%, 33/120) compared with CON (53.9%, 69/128). Pregnancy rate to TAI was less (P<0.01) in DESL-implanted cows that had initiated estrous cycles after calving (30.6%, 19/62) compared with CON (53.7%, 65/121) cows having initiated estrous cycles after calving. Furthermore, pregnancy rate was less (P<0.01) for cows having ovulations that had initiated estrous cycles after TAI in the DESL implant (39.1%, 18/46) compared with CON (62.1%, 54/87) treatments. Pregnancy losses from day 28 to day 46 of pregnancy did not differ between DESL implant (15.1%, 5/33) and CON (13.0%, 9/69) treatments. The DESL implant induced a delay in initiation of a new wave of follicular development during the postpartum-heat stressed period. The lesser pregnancy rate in the DESL-implant treatment group may be due to a pool of heat stress damaged follicles that were depleted in the control group with re-occurring follicle waves.     

Incorporation of a rapid pregnancy-associated glycoprotein ELISA into a CIDR-Ovsynch resynchronization program for a 28 day re-insemination interval

The objective was to compare two resynchronization programs; one that used a blood-based ELISA for pregnancy-associated glycoproteins (PAG) for pregnancy diagnosis so that non-pregnant cows were re-inseminated at 28 d after first TAI, and another that used transrectal ultrasonography for pregnancy diagnosis so that non-pregnant cows were re-inseminated at 35 d after first TAI. The PAG_resynch cows (n = 103) began CIDR-Ovsynch resynchronization on Day 18 after first TAI (Day 0). On Day 25, the CIDR was removed and pregnancy diagnosis with a PAG ELISA was performed. If a cow was not pregnant on Day 25, she was treated with PGF_2α, treated with GnRH 2 d later (Day 27), and TAI on Day 28. Control cows (n = 99) were observed for estrus until Day 25, when they began an identical CIDR-Ovsynch program with pregnancy diagnosis by transrectal ultrasonography on Day 32. If a cow was not pregnant on Day 32, then she was treated with PGF_2α, treated with GnRH 2 d later (Day 34), and TAI on Day 35. There was no difference in pregnancy per AI (P/AI) for either group at first or second insemination. For cows without pregnancy loss, the interval between first and second (P < 0.001) or second and third (P < 0.016) TAI was shorter for PAG_resynch cows compared with Control cows. The interval between first and second or second and third TAI was not different if pregnancy loss cows were included in the analysis. Plasma progesterone concentrations were similar at PGF_2α treatment, and plasma estradiol concentrations increased similarly after PGF_2α treatment for PAG_resynch and Control cows. In conclusion, the 28 d CIDR-Ovsynch resynchronization protocol was comparable to a 35 d CIDR-Ovsynch resynchronization protocol that also included estrus detection. Shortened resynchronization protocols that do not require estrus detection may improve reproductive efficiency in dairy cattle.     

Nutrient restriction and realimentation in beef cows during early and mid-gestation and maternal and fetal hepatic and small intestinal in vitro oxygen consumption

Objectives were to determine the effects of advancing gestation, maternal nutrient restriction during early and mid-gestation, and realimentation on fetal liver and jejunal mass and energy use in both dams and fetuses. On day 30 of pregnancy, multiparous, non-lactating beef cows (initial BW=621±11.3 kg and body condition score=5.1±0.1) were assigned to one of the two dietary treatments: control (CON; 100% requirements; n=18) and restricted (R; 60% requirements; n=28). On day 85, cows were slaughtered (CON, n=6; R, n=6), and remaining cows continued on control (CC; n=12) and restricted (RR; n=12) diets, or were realimented to the control diet (RC; n=11). On day 140, cows were slaughtered (CC, n=6; RR, n=6; RC, n=5), remaining cows continued on the control diet (CCC, n=6; RCC, n=5), or were realimented to the control diet (RRC, n=6). On day 254, all remaining cows were slaughtered. Maternal liver O₂ consumption linearly increased (P⩽0.04) and jejunal weight (g/kg) linearly decreased (P=0.04) as gestation advanced in CON groups. Fetal BW, and hepatic and small intestinal absolute mass, protein content and O₂ consumption linearly increased (P⩽0.04) as pregnancy advanced in CON groups. However, mass and O₂ consumption relative to BW linearly decreased (P⩽0.001) in the fetal liver in CON groups. When analyzing the effects of dietary treatment, at day 85, fetal jejunal O₂ consumption (mol/min per kg BW) was lower (P=0.02) in the R group when compared with the CON group. At day 140, maternal hepatic weight (g) was lower (P=0.02) in RC and RR cows when compared with CC, and fetal jejunual O₂ consumption (mmol/min per mg tissue and mmol/min per g protein) was greater (P⩽0.02) in RC when compared with RR. At day 254, maternal hepatic O₂ consumption (absolute and relative to BW) was lower (P⩽0.04) in the RCC cows when compared with RRC. Fetal hepatic weight was lower (P=0.05) in the CCC group when compared with RCC and RRC. The changes in response to nutrient restriction and realimentation in both the dam and fetus may indicate an adaptation to a lower amount of available nutrients by altering tissue mass and metabolism.     

Added dietary cobalt or vitamin B12, or injecting vitamin B12 does not improve performance or indicators of ketosis in pre- and post-partum Holstein-Friesian dairy cows

Vitamin B₁₂ is synthesised in the rumen from cobalt (Co) and has a major role in metabolism in the peri-paturient period, although few studies have evaluated the effect of the dietary inclusion of Co, vitamin B₁₂ or injecting vitamin B₁₂ on the metabolism, health and performance of high yielding dairy cows. A total of 56 Holstein-Friesian dairy cows received one of four treatments from 8 weeks before calving to 8 weeks post-calving: C, no added Co; DC, additional 0.2 mg Co/kg dry matter (DM); DB, additional 0.68 mg vitamin B₁₂/kg DM; IB, intra-muscular injection of vitamin B₁₂ to supply 0.71 mg/cow per day prepartum and 1.42 mg/cow per day post-partum. The basal and lactation rations both contained 0.21 mg Co/kg DM. Cows were weighed and condition scored at drying off, 4 weeks before calving, within 24 h of calving and at 2, 4 and 8 weeks post-calving, with blood samples collected at drying off, 2 weeks pre-calving, calving and 2, 4 and 8 weeks post-calving. Liver biopsy samples were collected from all animals at drying off and 4 weeks post-calving. Live weight changed with time, but there was no effect of treatment (P>0.05), whereas cows receiving IB had the lowest mean body condition score and DB the highest (P<0.05). There was no effect of treatment on post-partum DM intake, milk yield or milk fat concentration (P>0.05) with mean values of 21.6 kg/day, 39.6 kg/day and 40.4 g/kg, respectively. Cows receiving IB had a higher plasma vitamin B₁₂ concentration than those receiving any of the other treatments (P<0.001), but there was no effect (P>0.05) of treatment on homocysteine or succinate concentrations, although mean plasma methylmalonic acid concentrations were lower (P=0.019) for cows receiving IB than for Control cows. Plasma β-hydroxybutyrate concentrations increased sharply at calving followed by a decline, but there was no effect of treatment. Similarly, there was no effect (P>0.05) of treatment on plasma non-esterified fatty acids or glucose. Whole tract digestibility of DM and fibre measured at week 7 of lactation were similar between treatments, and there was little effect of treatment on the milk fatty acid profile except for C15:0, which was lower in cows receiving DC than IB (P<0.05). It is concluded that a basal dietary concentration of 0.21 mg Co/kg DM is sufficient to meet the requirements of high yielding dairy cows during the transition period, and there is little benefit from additional Co or vitamin B₁₂.     

Effect of pre-grazing herbage mass on dairy cow performance,grass dry matter production and output from perennial ryegrass (Lolium perenne L.) pastures

A grazing study was undertaken to examine the effect of maintaining three levels of pre-grazing herbage mass (HM) on dairy cow performance, grass dry matter (DM) production and output from perennial ryegrass (Lolium perenne L.) pastures. Cows were randomly assigned to one of three pre-grazing HM treatments: 1150 – Low HM (L), 1400 – Medium HM (M) or 2000 kg DM/ha – High HM (H). Herbage accumulation under grazing was lowest (P<0.01) on the L treatment and cows grazing the L pastures required more grass silage supplementation during the grazing season (+73 kg DM/cow) to overcome pasture deficits due to lower pasture growth rates (P<0.05). Treatment did not affect daily milk production or pasture intake, although cows grazing the L pastures had to graze a greater daily area (P<0.01) and increase grazing time (P<0.05) to compensate for a lower pre-grazing HM (P<0.01). The results indicate that, while pre-grazing HM did not influence daily milk yield per cow, adapting the practise of grazing low HM (1150 kg DM/ha) pasture reduces pasture DM production and at a system level may increase the requirement for imported feed.     

Evaluation of vaginal electrical resistance as an indicator of follicular maturity and suitability for timed artificial insemination in beef cows subjected to a synchronization of ovulation protocol

Objectives were to test the hypothesis that vaginal electrical resistance (VER) could be used to identify cows without a large (<10mm) follicle at timed-AI (TAI) following a synchronization of ovulation protocol and thus serve as a prospective decision aid for determining cows that should not be inseminated. Brahman x Hereford (F1) females (n=233) were synchronized with the CO-Synch+CIDR protocol that consisted of a controlled internal drug release (CIDR) insert and i.m. injection of GnRH (GnRH-1; 100 microg) on day 0, removal of CIDR and i.m. injection of prostaglandin F2alpha (PGF; 25mg) on day 7, and i.m. injection of GnRH (GnRH-2, 100 microg) and TAI 66h after CIDR removal (day 10). Vaginal electrical resistance was determined with a commercially available device (Ovascan; Animark Inc., Aurora, CO) at days 0, 7, and 10. Transrectal ultrasonography was used on day 10 to assess ovarian morphology at TAI in all cattle and in a subset of females (n=98) on days 0 and 7. Mean (+/-S.E.M.) age, body condition score (BCS), BW and days postpartum were 7.2+/-0.3 years, 5.2+/-0.1, 538+/-5.3kg, and 77+/-1.1 days, respectively. Mean VER (Omega) was greatest (101.4+/-0.8) on day 0 and declined (P<0.01) to 95.2+/-0.8 and 82.0+/-0.8 Omega, respectively, on days 7 and 10. Mean diameter of the largest follicle and VER values in females conceiving after TAI differed (P=0.05) from those that did not conceive. Mean VER on days 7 and 10 and VER difference (VER on day 10 minus VER on day 7) did not differ between females with a small (<10mm) or large (>or=10mm) follicle at TAI. Timed-AI pregnancy rate was greater (P<0.01) for females with large follicles (43%) than those with small follicles (22%). Vaginal electrical resistance difference values, categorized as negative (<0 Omega) or neutral/positive (>or=0 Omega), did not differ between females that conceived to TAI compared with those that did not. We conclude that VER measurements, as used in the present study, are not adequately sensitive to differentiate between females with and without a large follicle and thus are unable to serve as a prospective decision aid for determining suitability for TAI after synchronization.     

Effects of nutrient restriction on the metabolic profile of Bos indicus-influenced and B. taurus suckled beef cows

Recent research from our group demonstrated that Bos indicus-influenced suckled beef cows had greater resilience to withstand nutrient restriction and establish pregnancy compared with B. taurus cows exposed to the same conditions. To further understand these findings, differences in metabolic profile between these same B. indicus-influenced and B. taurus females were explored. Suckled beef cows (n = 134) were enrolled in a completely randomized design with a 2 × 2 factorial arrangement of treatments. On day − 21, Angus (AN; Bos taurus) and Brangus (BN; B. indicus-influenced) cows were randomly assigned to 1) a diet that met daily energy maintenance requirements (MAINT), or 2) a diet that restricted intake to 70% of the daily energy maintenance requirements (RESTR). Cows were exposed to an estrus synchronization protocol and received an embryo 7 d after ovulation was pharmacologically induced on day 0. Blood samples were collected on days − 21 and 19 to determine circulating concentrations of non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHB), insulin, glucose, and IGF-1. Pregnancy status after embryo transfer was determined on day 28. As a consequence of the proposed diets, cows in the RESTR diet had less body condition score (BCS) on day 19 (P = 0.008) across breed types. Moreover, BCS change from day − 21 to 19 was included as independent covariate into subsequent analyses, allowing for the comparison of breed types under an equivalent level of body reserve mobilization. A breed × diet interaction was observed for plasma insulin (P = 0.03) and IGF-1 (P = 0.04) on day 19, where AN–RESTR cows had less plasma concentrations on day 19 compared with AN–MAINT cows. Diets did not impact (P > 0.10) plasma insulin and IGF-1 concentrations in BN cows. No diet or breed effects were observed in circulating concentrations of NEFA, BHB, and glucose (P > 0.10). Across breed types and nutritional treatment, there was positive linear effect (P ≤ 0.04) of plasma concentrations of insulin and IGF-1 on the probability of pregnancy to fixed-time embryo transfer. In summary, the negative impacts of nutrient restriction on the somatotropic axis, independently of body tissue mobilization, were heightened in Bos taurus females compared with B. indicus-influenced cohorts, which corroborate with the differences observed in fertility between these subspecies.     

Effect of intermittent injections of gonadotrophin releasing hormone at various frequencies on the release of luteinizing hormone and ovulation in dairy cows

Gonadotrophin releasing hormone (GnRH, 5 μg every 4 h) was administered to six dairy cows between days 5 and 10 post-partum and the release of luteinizing hormone (LH) and the onset of ovulation were determined. LH was measured using a specific radioimmunoassay and the occurrence of ovulation was assessed from changes in the concentration of progesterone in milk. Treatment with GnRH resulted in a median time of first ovulation of 17.0 days after calving. This was less (P < 0.05) than that observed for control cows (21.5 days, n = 7). Determinations of plasma LH concentrations over an 8-h period on days 6 and 10 post-partum indicated that there was a tendency for GnRH-treated cows to have higher levels of LH on these days. The 5 μg dose of GnRH did not repeatably induce a release of LH between days 6 and 10. Endogenous pulsatile release of LH did, however, increase in frequency from 3.18 pulses per 8 h on day 6 to 5.18 pulses per 8 h on day 14 post-partum (P < 0.01).In a second experiment groups of 20 cows were treated with either 5 μg GnRH every 4 h or 15 μg GnRH every 12 h from days 5 to 10 post-partum. Seventeen untreated cows served as controls. The median times to first ovulation were 27.0 days for the control cows, 22.5 days for those cows treated with 5 μg GnRH every 4 h and 17.0 days for cows treated with 15 μg every 12 h. The latter treatment significantly advanced the time of first ovulation (P < 0.05) relative to controls. This difference had, however, disappeared by the time of the second and third ovulations. Primiparous cows ovulated later (P < 0.01) than the pluriparous cows in the group treated with 5 μg GnRH every 4 h. This was a major reason for the lack of effect of this treatment. Some treated cows were blood sampled at frequent intervals on day 8 to evaluate the LH responses to GnRH injections. The administration of 5 μg GnRH on day 8 did not elicit a pulse of LH which could be distinguished from endogenous pulsatile secretion at this time. The dose of 15 μg on this day did, however, elicit a more defined pulse on some, but not all, occasions.The injection of a small dose of GnRH twice a day from day 5 to day 10 after calving, therefore, advanced the time of first ovulation in dairy cows by 10 days.