W.W. Thatcher

Uterine-conceptus interactions and reproductive failure in cattle

The dialogue between trophectoderm cells of the conceptus and epithelial cells of the endometrium is critical to CL maintenance and embryo survival. The signal transduction mechanisms by which bovine interferon (IFN)-tau regulates cyclooxygenase (COX)-2 expression and secretion of prostaglandin F2alpha (PGF2alpha) in bovine endometrial (BEND) cells is examined. Stimulation of Protein Kinase C with a phorbol ester (phorbol 12, 13 dibutyrate [PDBu]) activates COX-2 gene expression and PGF2alpha secretion via the mitogen-activated protein kinase (MAPK) pathway. Interferon-tau attenuates PDBu activation of PGF2alpha secretion, but this inhibitory effect appears to be independent of the MAPK pathway. Embryonic IFN-tau, acting through a Type I IFN receptor, activates the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway resulting in activation or repression of interferon-stimulated genes. Experimental evidence is provided that IFN-tau regulation of STATs regulates gene expression of COX-2 in a manner that decreases secretion of PGF2alpha. Maternal regulation of the antiluteolytic pathway is discussed relative to the ability of the polyunsaturated fatty acid, eicosapentaenoic (EPA), to decrease endometrial secretion of PGF2alpha and progesterone to increase both conceptus development and IFN-tau secretion.

Embryonic development in superovulated dairy cattle exposed to elevated ambient temperatures between Days 1 to 7 post insemination

Holstein heifers (n = 29) were used to determine whether thermal stress during the first 7 d of embryonic development may increase the incidence of embryonic abnormalities in dairy cattle. Heifers were acclimated to environmental chambers at 20 degrees C for 9 d and superovulated with follicle stimulating hormone-pituitary (FSH-P; 40 mg total), beginning on Days 9 to 11 of the estrous cycle. Prostaglandin F(2)alpha (Lutalyse; 50 mg total) was administered on Day 3 of FSH-P. Heifers were inseminated artificially at estrus and then maintained at either thermal neutrality (20 degrees C) or under hyperthermic conditions (daily exposure up to 16 h at 30 degrees C and 8 h at 42 degrees C) for 7 d beginning at 30 h after the onset of estrus. On Day 7 post estrus, embryos were recovered nonsurgically and evaluated morphologically for stage of development and quality. The distribution of embryos classified as normal, abnormal, retarded or as unfertilized ova, differed (P<0.001) between heat stress and thermoneutral treatments. Only 20.7% of 82 embryos recovered from stressed heifers were normal compared with 51.5% of 68 embryos from thermoneutral animals. Stressed heifers had a higher incidence of abnormal and retarded embryos with degenerate nonviable blastomeres. Responses indicated that thermal stress from 30 h after the onset of estrus to Day 7 post estrus increases the incidence of abnormal and retarded embryos in superovulated heifers.

Effect of environmental heat stress on follicular development and steroidogenesis in lactating Holstein cows.

Lactating Holstein cows were utilized over two replicate periods (July and September, 1990) to examine the effect of summer heat stress on follicular growth and steroidogenesis. On day of synchronized ovulations, cows were assigned to shade (n=11) or no shade (n=12) management systems. Follicular development was monitored daily by ultrasonography until ovariectomy on Day 8 post estrus. At time of ovariectomy, dominant and second largest follicles were dissected from the ovary. Aromatase activity and steroid concentrations in dominant and subordinate follicles were measured. Acute heat stress had no effects on patterns of growth of first wave dominant and subordinate follicles between Days 1 and 7 of the cycle. Compared with shaded cows, the heat stressed cows did not have suppression of medium size (6 to 9 mm) follicles between Days 5 and 7. A treatment x follicle interaction was detected (P<0.01) for follicular diameter and fluid volume at Day 8. Dominant follicles in shade were bigger (16.4>14.5 mm) and contained more fluid (1.9>1.1 ml) than dominant follicles in no shade. Conversely, subordinate follicles in no shade were bigger (10.1>7.9 mm) and contained more fluid (0.4>0.2 ml) than subordinate follicles in shade. Concentrations of estradiol in plasma and follicular fluid were higher (P<0.01) in July than in September. Heat stress appears to alter the efficiency of follicular selection and dominance, and to have adverse effects on the quality of ovarian follicles.

Embryonic development in superovulated dairy cattle exposed to elevated ambient temperatures between the onset of estrus and insemination

Holstein heifers (n = 16) were used to determine whether heat stress prior to ovulation increases the incidence of embryonic abnormalities. Heifers were superovulated with Follicle Stimulating Hormone (FSH-P; 32 mg total), beginning on Days 10 or 11 of the estrous cycle. Prostaglandin F2α (Lutalyse; 60 mg total) was administered on Day 3 of FSH-P treatment. Heifers were maintained at either thermoneutrality (24°C) or under hyperthermic conditions (exposure to 42°C for 10 h) beginning at the onset of estrus. Following artificial inseminations at 15 and 20 h after the onset of estrus, heifers were continuously maintained under environmental conditions of thermoneutrality for 7 days as provided by environmental shade structures. On Day 7 post estrus, embryos were recovered nonsurgically and evaluated morphologically for stage of development and quality. The distribution of embryos classified as normal, retarded and/or abnormal, or as unfertilized ova differed (P < 0.001) between heat stress and thermoneutral treatments. Only 12.0% of 25 embryos recovered from heat-stressed heifers were normal compared with 68.4% of 19 embryos from thermoneutral heifers. Stressed heifers had a higher (P < 0.001) incidence of retarded and/or abnormal embryos with degenerated blastomeres. These data indicate that thermal stress during the periovulatory period increases the incidence of retarded and/or abnormal embryos in superovulated heifers.

New clinical uses of GnRH and its analogues in cattle

Abstract Gonadotropin-releasing hormone (GnRH) and its analogues cause an acute secretion of luteinizing hormone (LH) and follicle stimulating hormone (FSH) such that concentrations in peripheral blood are elevated for a 3–5 h period. GnRH-induced alterations in the function of the corpus luteum (CL) or follicle appear to be indirect through alterations in LH and FSH secretion. Repeated injections of GnRH during diestrus or single injections late in diestrus will cause acute increases in plasma progesterone and a delay in CL regression. Injections or continual administration of GnRH during early phases of CL development appear to augment CL differentiation and alter subsequent CL function. These effects are attributable to induced increases in LH. Injections of GnRH during the estrous cycle will re-synchronize follicle development owing to ovulation or luteinization of the dominant follicle leading to subsequent recruitment and selection of a new dominant follicle during a 7 day period. Injection of GnRH followed by injection of prostaglandin (PGF 2α ) at 6 or 7 days is a system of estrous synchronization in which follicle development and CL regression are both synchronized and fertility at the induced estrus is good. Injection of GnRH during the luteal phase post-insemination (e.g. Days 12–14) or post-embryo transfer, to alter CL and/or follicular function, has not resulted in a consistent increase in pregnancy rates. An overall assessment of studies that injected GnRH at the time of insemination in first service postpartum cows or in repeat breeders is rather disappointing. Considerable variation existed among studies within both types of cow populations relative to significant differences, directions of pregnancy rate change ( + vs. −), and magnitude of pregnancy rate increases. Recent findings indicate that timing of GnRH injections closer to the onset of estrus may be beneficial in increasing the conception rate. Utilization of GnRH in combination with progesterone and PGF results in an acute treatment sequence to program follicular development, ovulation and a subsequent cycle in cows with follicular cysts. The use of GnRH, with or without PGF, as a reproductive management program in the early postpartum period has not shown a clear improvement in subsequent reproductive efficiency. Development of precise systems to control ovarian function and reproductive efficiency with GnRH and other pharmaceutical agents is possible. However, such advancements must be founded on a clear understanding of GnRH-induced physiological effects and ability to capture any advantage by good management of the farm unit.

A study of prostaglandin F2α as the luteolysin in swine: II characterization and comparison of prostaglandin F, estrogens and progestin concentrations in utero-ovarian vein plasma of nonpregnant and pregnant gilts

Polyvinyl catheters were inserted into the right and left utero-ovarian veins (UOV) and saphenous vein (SV) and artery (SA) of six non-pregnant (O) and five pregnant (P) gilts on day 11 after onset estrus. Beginning on day 12, UOV blood samples were collected at 15-min intervals from 0800 to 1100 hr and 2000 to 2300 hr, and single samples were taken at 1200 and 2400 hrs. Peripheral blood (SA or SV) was sampled at 0800, 1200, 2000 and 2400 hr until gilts returned to estrus (X = 20.6 days) or day 24 of pregnancy. UOV plasma PGF concentrations (ng/ml; n = 1929) were measured by RIA. Status (P vs O) by day interactions were detected (P less than .01) but variances among treatments were heterogenous (P less than .01). Curvilinear day trends were detected for PGF in 0 gilts (P less than .01) but not P gilts. PGF peaks, defined as concentrations greater than two SD above the mean concentration for each gilt, occurred with greater frequency (chi2 = 16.4; P less than .01) in O than P gilts; and mean peak levels (X +/- SE) were 5.04 +/- .27 and 3.84 +/- .13 ng/ml, respectively. Progesterone concentrations were maintained in pregnant pigs and were indicative of luteal maintenance. Systematic differences in day trends of utero-ovarian venous plasma estradiol were detected between O and P pigs. These differences may be of paramount physiological importance and are discussed.

Long Chain Fatty Acids of Diet as Factors Influencing Reproduction in Cattle

Cattle are fed moderate amounts of long chain fatty acids (FA) with the objective to enhance lactation and growth; however, recent interest on lipid feeding to cows has focused on reproduction, immunity and health. Increasing the caloric density of the ration by fat feeding has generally improved measures of cow reproduction, but when milk yield and body weight losses were increased by fat supplementation, positive effects on reproduction were not always observed. Feeding fat has influenced reproduction by altering the size of the dominant follicle, hastening the interval to first postpartum ovulation in beef cows, increasing progesterone concentrations during the luteal phase of the oestrous cycle, modulating uterine prostaglandin (PG) synthesis, and improving oocyte and embryo quality and developmental competence. Some of these effects were altered by the type of FA fed. The polyunsaturated FA of the n-6 and n-3 families seem to have the most remarkable effects on reproductive responses of cattle, but it is not completely clear whether these effects are mediated only by them or by other potential intermediates produced during rumen biohydrogenation. Generally, feeding fat sources rich in n-6 FA during late gestation and early lactation enhanced follicle growth, uterine PG secretion, embryo quality and pregnancy in cows. Similarly, feeding n-3 FA during lactation suppressed uterine PG release, and improved embryo quality and maintenance of pregnancy. Future research ought to focus on methods to improve the delivery of specific FA and adequately powered studies should be designed to critically evaluate their effects on establishment and maintenance of pregnancy in cattle.

Seasonal and acute heat stress effects on steroid production by dominant follicles in cows

The present study concerned the seasonal and acute effects of heat stress on steroid concentrations in follicular fluid and on steroid production by granulosa and theca interna cells, in bovine dominant follicles. Three groups of cows were studied: summer (n = 5), autumn (n = 5) and winter (n = 9) cows. During the winter season, another group of cows was acutely heat-stressed from days 3 through 5 of the estrous cycle (n = 5). On day 7 of the estrous cycle, follicular fluid from first-wave dominant follicles was aspirated, and dispersed granulosa and theca cells from each seasonal group were incubated for 18 h at normothermic (37.5 degrees C) or high (40.5 degrees C) temperatures. Cells were incubated in media only or in media containing testosterone (300 ng ml-1, for granulosa cells) or forskolin (4 micrograms ml-1, for theca cells). In follicular fluid the 17 beta-estradiol concentration was high (P < 0.05) in winter and low in autumn, and summer, the androstenedione concentration was high in summer (P < 0.05), low in autumn, and intermediate in winter. During the winter season, acute in vivo heat stress increased follicular fluid androstenedione and decreased estradiol to levels comparable with those prevailing in summer. Basal and forskolin-stimulated androstenedione production by theca cells was higher (P < 0.05) in the winter group than in the summer and autumn groups, and also higher than in the cows that were heat-stressed during winter, which suggests that theca cell function is susceptible to chronic (summer), short-term (winter) and delayed (autumn) heat stresses. In vitro incubation at high temperature (40.5 degrees C) reduced the high, forskolin-stimulated androstenedione production in winter (P < 0.05). Estradiol production by granulosa cells was high in winter and autumn, and low in summer (P < 0.05). Acute heat stress in winter did not alter estradiol production relative to winter controls, whereas a high incubation temperature (40.5 degrees C) reduced (P < 0.05) estradiol production only in the autumn, when the highest production rate was recorded. The results indicate a differential effect of heat stress on the functions of granulosa and theca cells. Both concurrent and delayed effects of heat stress on the steroidogenic capacity of ovarian follicles in cattle are presented.

Effects of hormonal treatments on reproductive performance and embryo production

Developments in the use of drugs to improve reproduction and embryo production have focused on estrus and ovulation synchronization protocols and embryonic survival. Protocols for synchronization of ovulation eliminate the need for detection of estrus and allow timed insemination of all cows enrolled. Various estrogenic, progestational, GnRH and PGF2 alpha-like drugs are used to synchronize follicle development, CL regression and induction of ovulation. Strategies are discussed to optimize such programs to maximize herd pregnancy rates. Use of bovine Somatotrophin (bST) in combination with the Ovsynch protocol resulted in increased pregnancy rates, indicating possible effects on oocyte and embryonic development. Treatment of embryo donor cows with bST reduced the proportion of unfertilized oocytes and increased the number of transferable embryos. Furthermore, bST increased pregnancy rate when given to the recipient. Sub-luteal plasma progesterone concentrations after insemination have been associated with lower pregnancy rates. Injection of hCG on day 5 post-insemination resulted in induction of an accessory CL, increased plasma progesterone concentrations and increased conception rates. Strategies involving the use of sustained GnRH agonists to enhance CL development and alter follicular development are considered for future programs to enhance pregnancy rates.

Influence of summer heat stress on pregnancy rates of lactating dairy cattle following embryo transfer or artificial insemination

Lactating Holstein cows were used to determine if pregnancy rate from embryo transfer (n = 113) differed from contemporary control cows (n = 524) that were artificially inseminated (AI). Holstein heifers (n = 55) were superovulated with FSH-P (32 mg total) and inseminated artificially during estrus and subsequently managed under shade structures. On Day 7 post estrus, embryos were recovered, and primarily excellent to good quality embryos (90.3%) were transferred to estrus-synchronized lactating cows. Cows were managed under conditions of exposure to summer heat stress. Pregnancy status was determined by milk progesterone concentrations at Day 21 and palpation per rectum at 45 to 60 d post estrus. Pregnancy rates of cows presented for AI (Day 21, 18.0%; Days 45 to 60, 13.5%) were typical for lactating cows inseminated during periods of summer heat stress in Florida. Pregnancy rates of embryo recipient cows were higher (P<0.001) than those of control cows (Day 21, 47.6%; Days 45 to 60, 29.2%). Summer heat stress had no adverse effect on heifer superovulatory response, but it increased (P<0.05) the incidence of retarded embryos (</= 16 cells) and embryos graded as fair to poor quality. Increased pregnancy rate of recipient lactating cows indicates that the bovine embryo is sensitive to maternal heat stress during the first 7 d after estrus. Embryo transfer may bypass this period of embryonic sensitivity and provide an alternative to AI to partially circumvent heat stress-induced infertility in cattle.