Epigenetic change in IGF2R is associated with fetal overgrowth after sheep embryo culture.

Manipulation or non-physiological embryo culture environments can lead to defective fetal programming in livestock. Our demonstration of reduced fetal methylation and expression of ovine IGF2R suggests pre-implantation embryo procedures may be vulnerable to epigenetic alterations in imprinted genes. This highlights the potential benefits of epigenetic diagnostic screening in developing embryo procedures.

In vivo oocyte recovery and in vitro embryo production from bovine oocyte donors treated with progestagen, oestradiol and FSH.

The effect of treatment of donor cattle with progestagen and oestradiol or FSH on in vivo oocyte recovery and in vitro embryo production was studied. Forty-eight beefxFriesian cows formed eight replicates of six treatments in a 2 (no steroid versus steroid)x3 (none, single or multiple dose(s) of FSH) factorial design in which follicles were aspirated once weekly for 3 weeks. Oocytes were graded, washed, matured for 20-24h and then inseminated with frozen/thawed semen from a single sire followed by coculture on granulosa cell monolayers. Treatment with steroid had no significant effect on any follicular, oocyte or embryo production variate other than to reduce the number (P<0.05) and the diameter of large follicles>10mm (P<0.01) present at aspiration. FSH increased numbers of medium (6-10mm) and large follicles (P<0.01) and there was a corresponding decrease in the number of small follicles (2-5mm; P<0. 01). The total number of follicles at aspiration increased from 17. 7+/-1.60 for animals not treated with FSH to 23.6+/-1.97 following multiple dose treatment with FSH (P<0.05). Significantly, more follicles were aspirated following FSH treatment (no FSH 9.7+/-1.09, single dose FSH 13.6+/-1.30, multiple dose FSH 17.3+/-1.52; P<0.05) and numbers of oocytes recovered per cow per week increased (no FSH 4.1+/-0.76, single dose FSH 5.3+/-0.87, multiple dose FSH 5.9+/-0. 94) but the differences were not significant. Significantly, more good oocytes (Category 1) were recovered from animals treated with FSH (P<0.05). There was no overall significant effect of FSH on embryo production rate or the total number of transferable embryos produced but the number of transferable embryos was highest following administration of multiple doses of FSH. In conclusion, progestagen plus oestradiol 17beta treatment did not affect follicle, oocyte and embryo production of oocyte donors aspirated once per week. FSH treatment, however, significantly increased the number of follicles aspirated and Category 1 oocytes recovered. Multiple dose administration of FSH resulted in the production of the highest number of transferable embryos but this effect was not significant.

Fatty acid composition of lipids in immature cattle, pig and sheep oocytes with intact zona pellucida.

Cattle, pig and sheep oocytes isolated from healthy cumulus-oocyte complexes were pooled, within species, to provide samples of immature denuded oocytes with intact zona pellucida (n = 1000 per sample) for determination of fatty acid mass and composition in total lipid, constituent phospholipid and triglyceride. Acyl-containing lipid extracts, transmethylated in the presence of a reference penta-decaenoic acid (15:0), yielded fatty acid methyl esters which were analysed by gas chromatograph. Mean (+/- SEM) fatty acid content in samples of pig oocytes (161 +/- 18 micrograms per 1000 oocytes) was greater than that in cattle (63 +/- 6 micrograms; P < 0.01) and sheep oocytes (89 +/- 7 micrograms; P < 0.05). Of 24 fatty acids detected, palmitic (16:0; 25-35%, w/w), stearic (18:0; 14-16%) and oleic (18:1n-9; 22-26%) acids were most prominent in all three species. Saturated fatty acids (mean = 45-55%, w/w) were more abundant than mono- (27-34%) or polyunsaturates (11-21%). Fatty acids of the n-6 series, notably linoleic (18:2n-6; 5-8%, w/w) and arachidonic acid (20:4n-6; 1-3%), were the most abundant polyunsaturates. Phospholipid consistently accounted for a quarter of all fatty acids in the three species, but ruminant oocytes had a lower complement of polyunsaturates (14-19%, w/w) in this fraction than pig oocytes (34%, w/w) which, for example, had a three- to fourfold greater linoleic acid content. An estimated 74 ng of fatty acid was sequestered in the triglyceride fraction of individual pig oocytes compared with 23-25 ng in ruminant oocytes (P < 0.01). It is concluded that the greater fatty acid content of pig oocytes is primarily due to more abundant triglyceride reserves. Furthermore, this species-specific difference, and that in respect of polyunsaturated fatty acid reserves, may underlie the contrasting chilling, culture and cryopreservation sensitivities of embryos derived from pig and ruminant (cattle, sheep) oocytes.

Aberrant fetal growth and development after in vitro culture of sheep zygotes.

The effects of in vitro culture systems for sheep zygotes on subsequent fetal growth and development to day 61 and day 125 of gestation were studied. Zygotes recovered from superovulated Scottish Blackface ewes approximately 36 h after intrauterine insemination using semen from a single Suffolk sire were cultured for 5 days in (a) a granulosa cell co-culture system (co-culture); (b) synthetic oviductal fluid medium without serum (SOF-); and (c) synthetic oviductal fluid medium supplemented with human serum (SOF+). Control embryos were recovered from superovulated donor ewes at day 6 after oestrus. Embryos were transferred at day 6 to synchronous Scottish Blackface recipient ewes. In total, 146 gravid uteri were recovered, comprising 97 at day 61 (20 co-culture, 27 SOF-, 25 SOF+ and 25 control) and 49 at day 125 (13 co-culture, 8 SOF-, 6 SOF+ and 22 control) of gestation. Fetuses derived from co-cultured embryos were 14% heavier (P < 0.01) by day 61 of gestation than those derived from control embryos. Growth coefficients derived from the linear allometric equation logey = logea + b logex (where y = organ mass; x = fetal mass) were significantly greater (P < 0.05) for liver, heart, kidneys and plantaris muscle in fetuses derived from co-cultured embryos, and for liver in fetuses derived from SOF+ embryos than those for control fetuses. Fetuses derived from co-cultured embryos were 34% heavier (P < 0.001) and fetuses derived from SOF+ embryos were 18% heavier (P < 0.01) by day 125 of gestation than those derived from control embryos. Growth coefficients for liver and heart for fetuses derived from co-culture and SOF+ embryos were also significantly greater (P < 0.05) at this stage of gestation than those for control group fetuses. In contrast, allometric coefficients for these organs in fetuses derived from embryos cultured in SOF without serum supplementation were not different from those for controls. Excessive volumes of amniotic fluid (polyhydramnios) were observed in 23% of conceptuses derived from co-cultured embryos. In vitro embryo culture can significantly influence fetal growth and this study provides quantitative evidence of major shifts in the patterns of organ and tissue development.

In vivo oocyte recovery and in vitro embryo production from bovine donors aspirated at different frequencies or following FSH treatment.

The effects of frequency of follicular aspiration and treatment of donor cattle with FSH on in vivo oocyte recovery and in vitro embryo production were studied. Simmental heifers (n = 24) formed 8 replicates of 3 treatments in which oocyte donors were aspirated 1) once a week, 2) twice a week, or 3) once a week following treatment with FSH for 3 d prior to aspiration. Oocytes were graded, washed, matured for 20 to 24 h and then inseminated with frozen/thawed semen from a single sire, followed by co-culture on granulosa cell layers. Embryo development was observed until Day 7 after insemination. Significantly fewer follicles per heifer per week were counted (14.7+/-2.3 vs. 27.4+/-3.1 vs. 23.1+/-2.8) and aspirated (12.0+/-2.0 vs. 21.8+/-2.7 vs. 20.1+/-2.6) in heifers on the once-weekly than twice-weekly aspiration treatment (P<0.01) or on the once-weekly aspiration after FSH treatment (P<0.05). There were no significant differences between treatments in the total number of oocytes recovered per week (5.6+/-1.2 vs. 8.9+/-1.5 vs. 6.1+/-1.2), but significantly more oocytes per heifer per week recovered from animals treated with FSH were graded Category 1 (2.8+/-0.4), i.e., >4 layers good cumulus with a clear, even cytoplasm, than from animals aspirated once (0.9+/-0.2; P<0.01) or twice a week (1.5+/-0.3; P<0.05). The number of transferable morulae plus blastocysts produced per heifer per week was higher from animals aspirated twice a week (2.4+/-0.4; P<0.05) or once a week following FSH treatment (2.1+/-0.4; P<0.05) than from animals aspirated once a week without FSH treatment (1.0+/-0.3). In conclusion, FSH treatment of bovine oocyte donors aspirated once a week enabled a similar number of transferable embryos to be produced per donor week as aspiration twice a week without FSH treatment. These 2 treatments produced twice as many transferable embryos per donor week as aspiration once a week without FSH treatment.

Temporary exposure of ovine embryos to an advanced uterine environment does not affect fetal weight but alters fetal muscle development.

Embryo transfer techniques may result in fetuses that are heavier at birth and that have been described as highly muscled. The aim of this study was to investigate myogenesis in lambs derived from embryo transfer. Embryos were transferred at Day 3 (estrus = Day 0) to a 3 days-advanced uterine environment, maintained there for 3 days, recovered, and then returned to a synchronous (Day 6) uterus; these fetuses comprised the asynchronous group. Control animals were created by synchronous embryo recovery and single transfer at Day 3. Asynchronous transfer did not affect fetal weight or curved crown-rump length between 46 and 135 days of gestation. No differences were detected between groups at Days 110-135 with respect to muscle mass or protein, RNA, and DNA content. However, total muscle fiber number was significantly increased in plantaris muscles from the asynchronous groups at Day 110 and Day 125, suggestive of prolonged hyperplasia. In addition, the levels of Myf 5 protein and the secondary-to-primary fiber ratio were altered in plantaris muscle from the asynchronous group. The growth data are in contrast to previously reported findings. The results show that fetal myogenesis can be altered by very early events in embryogenesis and suggest that any inferences made solely on the basis of fetal or muscle weight may be fallacious.

Short-term culture of ovine embryos modifies fetal myogenesis.

Certain reproductive techniques culture embryos in vitro; however, little is known about the impact of culture on fetal growth. Coculture of day 1 ovine zygotes on a bovine granulosa cell layer to blastocysts followed by transfer to synchronous recipients increased fetal weight by 11 and 40% at days 61 and 125, respectively, compared with the transfer of in vivo-produced blastocysts. Plantaris muscle weights were increased by 40% in cultured fetuses at day 125. Examination of myogenesis in plantaris muscle showed that primary fiber number was unchanged at day 61 by culture but that primary fiber area was increased significantly by 15 and 25% at days 61 and 125, respectively; secondary fiber area was increased by 40% at day 125 by culture, and the ratio of secondary to primary fiber numbers was 18-20% greater in the cultured groups compared with the controls at days 61 and 125. The results show that coculture of preimplantation embryos may alter myogenic programming. These changes may contribute to the abnormally large muscles observed in oversize fetuses.

Post-natal growth and development of Simmental calves derived from in vivo or in vitro embryos.

Large fetuses arising from embryos produced in vitro have been shown to exhibit altered organ development in utero, but it is not known whether this persists post natally. Post-natal growth and development was examined in 18 Simmental bulls derived from in vivo frozen-thawed (n = 6), in vitro frozen-thawed (n = 6) or in vitro fresh (n = 6) embryos and reared together post weaning on an ad libitum diet until slaughter at approximately 13 months old. Calves weighing less than 60 kg at birth (n = 11) were classified as normal, and heavier calves (n = 7; all from in vitro embryos) as oversize. Lifetime growth rates and slaughter weights apparently were unaffected by embryo source or birthweight. Mean (+/- s.e.m.) post mortem liver and kidney weights were unaffected by embryo source, but hearts of bulls from in vitro frozen embryos were heavier than those of bulls from in vivo frozen embryos (2.7 +/- 0.04 v 2.3 +/- 0.07 kg, P<0.025). Heart weight per kilogram body weight at slaughter for the 7 perinatally oversize males (4.01 +/- 0.08 g) exceeded that of the other 5 bulls from in vitro embryos (3.60 +/- 0.10 g kg(-1); P<0.04) and the 6 in vivo males (3.56 +/- 0.12 g kg(-1); P<0.02). Overall, one-third of the variation in heart weight at slaughter (r2 = 0.35; P = 0.01) was due to variation in birthweight. This is the first study to demonstrate birthweight-related developmental effects on post-natal organ weight following the transfer of embryos produced in vitro.

Fetal growth and development following temporary exposure of day 3 ovine embryos to an advanced uterine environment.

The effect of exposing Day 3 ovine embryos to an advanced uterine environment for a period of 3 days on subsequent fetal growth and development between Day 35 and Day 135 of gestation was studied. Day 3 embryos were recovered from superovulated donor ewes and transferred to synchronous final or asynchronous temporary recipients for 3 days. Embryos were recovered from these temporary recipients and transferred to Day 6 final recipients. Gravid uteri were recovered, weighed and dissected on Days 35, 45, 60, 90, 110, 125 and 135 of gestation. Fetal weight and length data were analysed by fitting non-linear Gompertz models of the form log(e) y = a - be(-ct), where y is fetal size and t is time from conception. Various terms including treatment, gestational age, embryo stage at transfer and fetal sex were fitted to this model. Fetal development was assessed by relating organ weight to fetal bodyweight using the linear allometric equation log(e) y = log(e) a + b log(e) x, where y is organ weight and x is fetal weight. Temporary exposure of Day 3 embryos to an advanced uterine environment did not increase the rate of embryo development and had no effect on fetal growth and development between Days 35 and 135 of gestation in this study. A single Gompertz model (log(e) y = 10.134 - 17.047e(-0.1733t)) explained 99.8% of the variation in fetal weight. Of terms fitted to this model only gestational age and fetal sex influenced fetal weight, with male fetuses being 5% heavier (P<0.05) than female fetuses. Fetal development was also unaffected by experimental treatment in this study. Allometric coefficients established for various fetal components agreed well with those from previously published studies.

Effect of frequency of follicle aspiration on oocyte yield and subsequent superovulatory response in cattle.

The effect of frequency of transvaginal follicular aspiration on oocyte yield and subsequent superovulatory response was studied in 2 experiments. In Experiment 1, 32 primiparous Hereford x Friesian cows were assigned to 4 treatments (n = 8 per treatment). Oocyte recovery was carried out once a week for 12, 8, 4 or 0 (control) wk. Embryo recovery for all animals was 7 wk after the completion of the aspiration schedules. In Experiment 2, the effects of oocyte recovery once or twice a week (n = 8 per treatment; control n = 18) for 12 wk and response to superovulation 4 wk after the last aspiration were compared using nulliparous purebred Simmental heifers. Increasing the period of once weekly aspirations from 4 to 12 wk (Experiment 1) did not affect the number of follicles observed per session (mean +/- SEM; 10.0 +/- 0.82) or aspirated (7.8 +/- 0.71), but the recovery rate of oocytes from follicles aspirated was greater for donors aspirated for either 4 or 8 wk than for 12 wk (32.3 +/- 3.73 vs 28.4 +/- 2.61 vs 20.1 +/- 2.13 %; P < 0.05). Following the last aspiration and prior to commencing superovulatory procedures, estrus or estrous activity was observed in 7 8 , 8 8 , 7 8 and 6 8 of the animals aspirated over 12, 8, 4 or 0 wk, respectively. Subsequent superovulatory responses and in vivo embryo recoveries were similar for all aspiration treatments and for control animals. Changing the frequency of oocyte recovery from once to twice weekly (Experiment 2) did not affect the numbers of follicles observed (9.1 +/- 0.63 vs 8.3 +/- 0.85), follicles aspirated (5.9 +/- 0.56 vs 6.2 +/- 0.69), oocytes recovered (1.7 +/- 0.27 vs 1.9 +/- 2.0) per session or the oocyte recovery rate (29.4 +/- 2.4 vs 30.4 +/- 2.4 %); nor was there any effect of frequency of aspiration on subsequent superovulatory response and embryo recovery. In conclusion, increasing the period of aspiration from 4 to 12 wk and the frequency from once to twice a week over 12 wk did not reduce the number of follicles observed or aspirated, or number of oocytes recovered per donor per session. Subsequent estrous cyclicity and responses to superovulation were unaffected by the periods or frequencies of oocyte recovery examined here.

The response of bovine granulosa cells to different gonadotrophins in culture.

Previous studies with bovine granulosa cells cultured in vitro indicated that follicle-stimulating hormone (FSH) stimulated differentiation and progesterone production of granulosa cells in a dose-dependent manner, this was due mainly to an increase in the number of differentiated cells. The objectives of the present study were to investigate (1) whether the response of bovine granulosa cells in culture to luteinising hormone (LH) and equine chorionic gonadotrophin (eCG) was similar to the response to FSH, and (2) whether granulosa cells derived from different cattle breeds responded similarly to gonadotrophin stimulation. Pairs of ovaries were recovered postmortem from Charolais (38) and Hereford (41) crossbred post-pubertal heifers, and granulosa cells were aspirated from 5-8 mm follicles. In two simultaneous experiments, granulosa cells (2-3 x 10(5) viable cells) were cultured with different gonadotrophins (oFSH or oLH in Experiment 1; oFSH or eCG in Experiment 2). Cell culture was for 4 days at 37 degrees C in a humidified atmosphere of 5% CO2 in air in 1 ml of serum-free culture medium. Progesterone production, total DNA and the protein content of granulosa cells on Day 4 of culture were determined. Log10 data were analyzed by analysis of variance and multiple linear regression. In Experiment 1, both FSH and LH stimulated progesterone production (ng microgram-1 DNA) and protein content (microgram microgram-1 DNA) of granulosa cells in a dose-dependent manner (P < 0.01). The relative potencies of FSH to LH (milli micron/milli micron) were found not to be different from unity. In Experiment 2, progesterone production and the protein content of granulosa cells were stimulated by both FSH and eCG in a dose-dependent manner (P < 0.001). The progesterone response curves (log/log) were linear up to 1-10 milli microns FSH and 1-10 iu eCG, and were Y = 1.67 + 0.093 FSH and Y = 1.60 + 0.091 eCG for progesterone production. Calculated on a milli micron/iu basis, FSH was found to be 5.8 times more potent than eCG (P < 0.05) in terms of stimulating progesterone production. Granulosa cells derived from Hereford crosses were more sensitive (P < 0.001) than those from Charolais crosses to gonadotrophin stimulation (31 and 42 times for FSH and eCG, respectively, in terms of progesterone production, and 4.8 and 3.1 times for FSH and eCG, respectively, in terms of protein content). The response curves for both FSH and eCG were similar within each breed. The slopes of the progesterone response curves, and the protein responses were similar for all the gonadotrophins. In conclusion, these results imply that FSH; LH and eCG have similar effects on the differentiation and progesterone production of bovine granulosa cells from 5-8 mm follicles cultured in vitro. Furthermore, granulosa cells from different breeds cultured in vitro had different sensitivities to gonadotrophin stimulation.

Increasing the efficiency of suckled calf production using embryo transfer technology.

The data reviewed in this paper illustrate the benefits of increased output that can be obtained from suckler herds using embryo transfer technology. The technology can be used within breeding schemes to increase the rate of genetic progress for selected traits or to transfer embryos of superior genetic merit and, in the future, embryos of predetermined sex to beef cows. The success of the technology is dependent on the achievement of good pregnancy rates. Experience gained on commercial farms suggests that the main reason for poor success rates in some herds lies with the general level of management of such herds rather than with the reproductive technology itself. Much experience has been gained on the management of embryo transfer recipients and twin-bearing cows. In particular, the nutritional requirements of such animals during the early post partum period and during mid and late pregnancy, and the management of twin-bearing cows during the perinatal period, are discussed.

Norgestomet implants, plasma progesterone concentrations and embryo transfer pregnancy rates in cattle.

A study was undertaken to test the hypothesis that supplementation with exogenous progestagen at the time of embryo transfer would enhance pregnancy rates in recipients. Two-hundred-and-seventy-two oestrus-synchronised crossbred heifer and cow recipients received 200 grade 1 and 72 grade 2 Simmental embryos transferred non-surgically. Heparinised blood samples were taken on day 6 and day 7 (oestrus = day 0) for the assessment of the endogenous plasma progesterone concentration. Half the recipients received an ear implant impregnated with 3 mg norgestomet on the day of embryo transfer. The pregnancy rates were 51.9 and 49.6 per cent for the norgestomet-treated and control groups, respectively. The pregnancy rate for grade 1 embryos was 56.0 per cent and for grade 2 embryos 36.1 per cent (P < 0.01). The breed of recipient, weekday of transfer, operator and condition score had no effect on pregnancy rate. The maiden heifers had a higher pregnancy rate (54.2 per cent) than the cows (46.2 per cent). The mean plasma progesterone concentrations of the pregnant and non-pregnant groups on day 6 were 6.7 ng/ml and 6.6 ng/ml, respectively, and 7.6 ng/ml in both groups on day 7.

Bovine granulosa cell culture for assessment of potency and specificity of antibodies to pregnant mares' serum gonadotrophin.

Antibodies to pregnant mares' serum gonadotrophin (PMSG) neutralize the effect of PMSG in vivo and increase the number of transferable embryos when administered at the optimum time relative to the preovulatory luteinizing hormone (LH) surge in PMSG-stimulated cows. The objective of the present study was to investigate the possible use of bovine granulosa cells in a serum-free culture system as a bioassay for antibodies to PMSG. Granulosa cells (2-3 x 10(5) viable cells) were cultured with varying doses of PMSG and/or an anti-PMSG for 4 days. Whilst progesterone production (ng/micrograms DNA) of granulosa cells was stimulated by PMSG (p < 0.01) in a dose-dependent manner, increasing amounts of anti-PMSG neutralized (p < 0.01) this stimulatory effect of either follicle-stimulating hormone or LH on progesterone production of bovine granulosa cells in vitro. The bovine granulosa cell culture system is a potential in vitro bioassay method for testing the specificity and the neutralizing capacity of different anti-PMSG preparations.

Evaluation of Crestar, a synthetic progestogen regime, for synchronising oestrus in maiden heifers used as recipients of embryo transfers.

Crestar consists of an ear implant containing 3 mg norgestomet combined with an intramuscular injection of 3 mg norgestomet and 5 mg oestradiol valerate. Its effectiveness for synchronising oestrus in embryo transfer recipients was evaluated in comparison with a progesterone-releasing intravaginal device (PRID) and prostaglandin regimen, using 334 maiden heifers. The treatment devices were inserted on day 1, prostaglandin was administered to the PRID-treated heifers on day 8 and the devices were removed on day 10. High proportions of the heifers were seen in oestrus within five days of the removal of the devices after both the PRID prostaglandin (90.4 per cent) and Crestar (86.2 per cent) treatments. The interval from the removal of the device to the onset of oestrus was significantly shorter for Crestar than for PRID prostaglandin-treated heifers (45 vs 51 hours, P < 0.001), and the duration of oestrus was significantly longer (13 vs 10 hours, P < 0.01). The PRID prostaglandin treatment resulted in a higher degree of synchrony than the Crestar treatment (74.1 per cent vs 61.8 per cent, P < 0.05). There were no significant differences between the treatments in the proportions of the heifers selected as embryo transfer recipients or in the proportions which became pregnant after embryo transfer.

Synchronization of estrus in embryo transfer recipients after using a combination of PRID or CIDR-B plus PGF2alpha.

The use of CIDR-B or PRID in combination with prostaglandin F2alpha (PGF2alpha) for synchronizing estrus in embryo transfer recipients was evaluated in 2 experiments. In Experiment 1, virgin heifers (n=263) were synchronized using either a PRID (including estradiol benzoate capsule) or a CIDR-B in a combined program in which devices were inserted on Day 1, an injection of prostaglandin was given on Day 6, and devices were withdrawn on Day 7. The interval from device removal to the onset of estrus was significantly shorter for CIDR-B than for PRID-treated animals (50.44 vs 55.50 hours; P<0.003). The CIDR-B treatment resulted in a similar degree of synchrony to the PRID treatment (74.0 vs 70.4%; P=0.68). InExperiment 2, cows (n=95) and heifers (n=93) were allocated at random to be synchronized using a PRID (excluding estradiol benzoate capsule) plus PGF2alpha or a CIDR-B device plus PGF2alpha. The devices were inserted on Day 1, an injection of prostaglandin was given on Day 10 and the devices were removed on Day 12. Estrus was observed earlier following the CIDR-B treatment (43.50 vs 47.04 hours; P=0.01), but the degree of synchrony was similar (76.2 vs 76.3%; P>0.10) for the CIDR-B and PRID-treated animals. In both experiments, there were no significant differences in the proportions of animals observed in estrus, selected as embryo transfer recipients, or which achieved pregnancy consequent on embryo transfer between those synchronized using CIDR-B or PRID regimens. We conclude that the CIDR-B is a suitable device for synchronizing estrus in embryo transfer recipients.

Changes in the composition of cervical mucus of the cow during the estrous cycle as parameters for predicting potential fertility.

Four experiments were conducted to test the hypothesis that the composition of cervical mucus can be used as an indicator of reproductive efficiency in the cow. In Experiment 1, biochemical changes were studied in cervical mucus during the estrous cycle. Sorbitol concentration was observed to be highest at 1 to 3 days prior to estrus and lowest on Days 6 to 12 (P<0.001) of the estrous cycle. Cholesterol and protein concentrations were highest at Day 6 of the estrous cycle and lowest on the day of estrus (P<0.001). In Experiment 2, the relationships between the biochemical characteristics of cervical mucus and fertility were studied. It was shown that the embryo transfer recipients which exhibited a high concentration of sorbitol (>1.5 mMol/l) at 1 to 3 days before estrus; a low concentration of protein (< 2 units); and a low concentration of cholesterol (<0.1 mMol/l) on the day of estrus had a higher level of fertility than their counterparts. The predictive ability of these criteria was tested using embryo transfer recipients (n=294) in Experiment 3. Significantly more of the animals predicted to have high potential fertility became pregnant than those predicted to have low potential fertility (70.7 vs 45.6%; P<0.001). A similar difference in pregnancy rate for cows (n=56) presented for artificial insemination was observed in Experiment 4 (59.1 vs 27.2%; P<0.10). These results suggest that the composition of cervical mucus may be a useful indication of potential fertility in cattle.