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Effect of dietary protein on embryo recovery rate and quality in superovulated heifers.

For almost 3 decades, superovulation and embryo transfer have been used in cattle breeding to increase the number of offspring from genetically superior female animals. Several factors including nutrition affect the number of transferable embryos recovered. We compared the effects of two different dietary protein levels easily achieved in practical conditions on embryo number and quality in superovulated heifers. Finnish Ayrshire heifers (n = 37) were allocated to isoenergic diets containing either 14% (D14) or 18% (D18) crude protein (CP). Estruses were synchronized, and the heifers were subsequently superovulated and inseminated using a standard FSH-protocol. Embryos were collected 7 days after inseminations (71-72 days after the beginning of the treatment period) by uterine flushing. The number of corpora lutea, and the number and quality of embryos were determined. Protein feeding did not affect superovulatory response, the number of embryos or the number of transferable embryos recovered. Proportionally more poor-quality embryos were found in group D14 than in group D18 (20.2% versus 13.2%, respectively, P = 0.053). It is concluded that a long-term moderate increase in the content of crude protein fed to energy-adequate heifers does not seem to affect superovulatory response and the number of embryos recovered, but it may be advantageous to the quality of embryos.

Animals↗

Pregnancy rates in mares following hysteroscopic or transrectally-guided insemination with low sperm numbers at the utero-tubal papilla.

This study was conducted to evaluate two methods for insemination of a low number of sperm in the tip of the uterine horn, and to determine whether prebreeding intrauterine treatment with prostaglandin E(2) would improve pregnancy rates. Estrus was synchronized in 36 fertile Quarter Horse and Thoroughbred broodmares. When a dominant follicle >or=33 mm diameter was present, mares were treated with 2500 units hCG intravenously and were assigned to one of four treatment groups for insemination with five million total sperm in 200 microl extender the next day as follows: (1) Group PGE-HYS (n=9): 0.25mg PGE(2) in 1 ml 0.9% NaCl solution infused into the tip of the uterine horn ipsilateral to the dominant follicle 2h prior to hysteroscopic-guided inseminate placement onto the oviductal papilla; (2) Group SAL-HYS (n=9): 1 ml 0.9% NaCl solution infused into the tip of the uterine horn ipsilateral to the dominant follicle 2h prior to hysteroscopic-guided inseminate placement onto the oviductal papilla; (3) Group PGE-PIP (n=9): 0.25mg PGE(2) in 1 ml 0.9% NaCl solution infused into the tip of the uterine horn ipsilateral to the dominant follicle 2h prior to transrectally-guided pipette placement of the inseminate into the tip of the uterine horn; and (4) Group SAL-PIP (n=9): 1 ml 0.9% NaCl solution infused into the tip of the uterine horn ipsilateral to the dominant follicle 2h prior to transrectally-guided pipette placement of inseminate into the tip of the uterine horn. Mares in estrus were evaluated daily by transrectal ultrasonography to monitor follicular status and confirm ovulation. If mares had not ovulated within 2 days of insemination, the assigned treatment was repeated. Pregnancy status was evaluated by transrectal ultrasonography 12-14 days postovulation, and pregnancy rates were compared. No interaction between prebreeding treatment (SAL:PGE) and insemination protocol (HYS:PIP) on pregnancy rates occurred (P>0.10). Pregnancy rates did not differ between mares inseminated by HYS (12/18; 67%) or PIP (10/18; 56%) (P>0.10). Pregnancy rates did not differ between mares treated prior to breeding with PGE (11/18; 61%) or SAL (11/18; 61%) (P=1.00). In summary, satisfactory pregnancy rates were obtained when a low number of sperm were either placed directly onto the oviductal papilla using hysteroscopy or placed in the tip of the uterine horn using a transrectally-guided uterine pipette. Infusion of 0.25mg PGE(2) in the tip of the uterine horn 2h prior to insemination did not improve pregnancy rates.

Animals↗

Birth of pouch young after artificial insemination in the tammar wallaby (Macropus eugenii).

Timing of artificial insemination (AI) in marsupials is critical because fertilization must occur before mucin coats the oocyte during passage through the oviduct. In this study, timing and the site of insemination were examined to develop AI in the tammar wallaby (Macropus eugenii). Birth and postpartum (p.p.) estrus was synchronized in 46 females. Epididymal spermatozoa (n=4) or semen collected by electroejaculation (n=42) were inseminated early (4-21 h p.p.) into the urogenital sinus (n=7), the anterior vaginal culs de sac (n=7), the uterus by transcervical catheter (n=5), or the uterus by injection (intrauterine artificial insemination, IUAI) (n=5). A further 16 females were inseminated late (19-48 h p.p.) by IUAI. All females were monitored for birth. A third group of six females was inseminated late (21-54 h p.p.) by IUAI and 0.4-6.6 h later, sperm had reached the oviduct in all animals. In total, an oocyte to which spermatozoa were attached was recovered and two young were born after IUAI using epididymal (n=1) or electroejaculated (n=2) spermatozoa, but no young resulted from insemination at other sites. Two females were successfully inseminated at 43 and 47 h p.p., later than most other animals, and the third was inseminated much earlier (18 h p.p.) but with highly motile spermatozoa. These young represent the first macropodids born by AI and the first marsupials conceived using epididymal spermatozoa.

Animals↗

Oxytocin-induced cervical dilation and cervical manipulation in sheep: effects on laparoscopic artificial insemination.

The difficulty of cervical penetration severely limits the use of transcervical AI (TAI) in sheep, and trauma from cervical manipulation (CM) may reduce fertility after TAI. We investigated the effects of cervical dilation using exogenous oxytocin (OT) to facilitate TAI and its effects on reproductive variables after laparoscopic AI (LAI). Estrus was synchronized by inserting pessaries impregnated with 6alpha-methyl-17alpha-hydroxyprogesterone acetate (60 mg) for 12 d. In Exp. 1, we determined whether OT and CM before LAI affected the interval from pessary removal to ovulation and fertilization rate. Crossbred ewes (n = 16) were assigned to 1) saline-CM or 2) OT-CM. In Exp. 2, effects of OT and CM on lambing rates were evaluated with white-faced ewes (n = 220) in a 2 x 2 factorial experiment: 1) saline-sham CM; 2) saline-CM; 3) OT-sham CM; and 4) OT-CM. In both studies, eCG (400 IU i.m.) was injected at pessary removal, and LAI was performed 48 to 52 h later. In Exp. 1, ewes received i.v. either 400 USP units of OT or 20 mL of saline at 30 to 60 min before LAI, and CM was administered as for TAI. Beginning 32 h after pessary removal and continuing at 8-h intervals, ovaries were examined with ultrasonography to estimate time of ovulation. Treatment in Exp. 1 did not affect combined ovum/embryo recovery rate (69%), but OT-CM decreased fertilization rate (47 vs 59%; P < 0.05). The OT tended to reduce the interval to ovulation (OT, 59 h vs saline, 66 h; P < 0.06). The OT x CM interaction in Exp. 1 was not significant. For Exp. 2, approximately 25 min before sham CM or CM, 200 USP units of OT or 10 mL of saline was injected i.v. The LAI was performed immediately after sham CM or CM. At 10 to 12 d after AI in Exp. 2, ewes were mated with Suffolk rams. Blood was collected between 24 and 26 d after AI for pregnancy-specific protein B (PSPB) RIA. The PSPB pregnancy and lambing rates were both 62% in saline-sham controls. The CM did not affect pregnancy (69%) or lambing rate (64%). The OT treatment decreased (P < 0.05) PSPB pregnancy (59%) and lambing rates (56%) in OT-sham ewes and pregnancy and lambing rates in CM ewes (both 43%). Neither CM nor OT before LAI affected lambing rates to next estrus, indicating no long-term damage to the cervix or uterus. In summary, CM did not affect fertility after LAI, but OT decreased lambing rate independent of CM. If OT will not be usable for TAI, it may still be a tool for training TAI personnel.

Animals↗

Induction of the presence of corpus luteum during superovulatory treatments enhances in vivo and in vitro blastocysts output in sheep.

This report offers the results of two experiments developed to test possible benefitial effects of the presence of corpus luteum (CL) on in vivo and in vitro sheep embryo production; using two different breeds treated with two different protocols by two different teams at two different centres. In the first trial, estrus was synchronized in 11 ewes with two doses of cloprostenol, 10 days apart. On day 1 after estimated ovulation, sheep were treated with progestagen sponges and superovulated with eight decreasing doses (26.4 units NIH-FSH-S1 x 3, 22.0 units x 2, and 17.6 units x 3) of ovine FSH injected twice daily. Ovulation rate and number of embryos obtained in vivo were compared to those from 12 control ewes without cloprostenol treatment. Presence of a CL improves the number of transferable embryos (7.4+/-0.6 versus 4.1+/-0.6 in control ewes, P < 0.05). The second trial investigated the effects of the presence of CL on embryos produced in vitro from six ewes bearing CL and six ewes without CL at start a superovulatory treatment consisting of 96 units of ovine FSH administered in four equal doses given every 12 h. There were not detected effects of the CL on the number and size of follicles or in the number, morphology and ability to resume meiosis of their oocytes. However, oocytes from ewes with CL showed higher rates of fertilization (73.5 versus 45.5%, P < 0.005), higher development to blastocyst (35.8 versus 19.3%, P < 0.01) and higher hatching rates after vitrification (80.0 versus 25.0%, P < 0.05).

Animals↗

Pregnancy and bovine somatotropin in nonlactating dairy cows: I. Ovarian, conceptus, and insulin-like growth factor system responses.

Nonlactating dairy cows were used to examine effects of bovine somatotropin (bST) on components of the insulin-like growth factor (IGF) system. Estrus was synchronized in cows with a Presynch + Ovsynch protocol and timed AI (TAI; n = 55) or not TAI (cycling, C; n = 23) on d 0 (time of synchronized ovulation). On d 0 and 11, cows received bST (500 mg) or no bST, and were sacrificed on d 17. Pregnancy rates were less in bST cows (27.2%, 9 of 33) than in controls (63.6%; 14 of 22). In contrast, conceptuses were larger in bST-treated cows (39.2 +/- 4.8 cm) than in controls (20 +/- 4.3 cm). Total interferon-tau in uterine luminal flushings (ULF) was greater in bST-treated cows (7.15 > 2.36 microg). Number of class 2 follicles (6 to 9 mm) was less in bST-C cows on d 7 and 16. On d 17, corpus luteum (CL) weight tended to be greater in bST-treated cows. Concentrations of progesterone were greater after d 10 in C than in pregnant (P) cows. In the ULF, IGF-binding protein-3 was greater in bST-P cows than in pregnant cows. A tendency for an increase in IGF-I hormone concentrations in the ULF was detected on d 17 in bST-treated and cyclic cows. Endometrial mRNA for IGF-I, IGF-II, IGFBP-2, and IGFBP-3 increased in bST-C, but not in bST-P cows. Treatment with bST increased plasma concentrations of insulin, IGF-I, and growth hormone (GH). In conclusion, bST may have hyperstimulated plasma IGF-I and insulin to cause asynchrony between conceptus and uterus that was detrimental to pregnancy.

Animals↗

Direct effects of luteal regression on anterior pituitary response to GNRH.

The preovulatory period of the ewe is marked by a dramatic decrease in concentrations of progesterone in serum during the late luteal phase, followed by elevated luteinizing hormone (LH) secretion, final follicular maturation and ovulation. This experiment was designed to ascertain the extent to which removal of endogenous progesterone negative feedback at the anterior pituitary gland, independent of effects at the hypothalamus, promotes increased secretion of LH in the hours immediately after induction of luteolysis. Estrus was synchronized in ovary-intact ewes with two injections of prostaglandin F2 alpha (PGF2 alpha) analog given 10 d apart (Day 0 = second day after the second PGF2 alpha injection). Ewes were subjected to hypothalamic-pituitary disconnection (HPD; n = 6) on Day 3 and were pulsed with gonadotropin-releasing hormone (GnRH). Ewes were used during the estrous cycle or received approximately 400 IU pregnant mare serum gonadotropin (PMSG) on Day 2 to stimulate ovulation; there was no difference (P < 0.10) in ovulation rate or progesterone production between these two groups. Luteal regression was induced by injection of PGF2 alpha analog on approximately Day 10 of the estrous cycle. Blood samples were collected around exogenous GnRH pulses before and at 2- or 4-hr intervals after PGF2 alpha administration and concentrations of LH and progesterone determined. At 4, 12, and 24 hr after PGF2 alpha administration, mean serum progesterone levels in all ewes had decreased by 54.7%, 66.2% and 89.4%, respectively (P < 0.05) from pre-injection levels. The decrease in progesterone was associated with an increase (P < 0.01) in LH pulse amplitude with means at 4-hr post-PGF2 alpha ranging from 190% to 288% of pre PGF2 alpha values. Mean serum LH levels were also increased (P < 0.01) within 4 hr of PGF2 alpha administration and remained elevated at all but the 24-hr time point. The timing of this increase (within 4 hr) indicates that it is independent of changes in serum estradiol concentrations, which do not increase for at least 16 hr after induction of luteolysis. Thus, removal of endogenous progesterone negative feedback at the anterior pituitary gland in the hours immediately after induction of luteolysis seems to play a role in facilitating LH release independently of hypothalamic action.

Animals↗

Ovarian follicular responses to high doses of pulsatile luteinizing hormone in lactating dairy cattle.

Two experiments in lactating dairy cows examined ovarian follicular responses to high, frequent doses of exogenous LH pulses at levels associated with follicular cysts. In Experiment 1, estrus was synchronized in 12 cyclic lactating cows >40 d postpartum. Emergence of the second follicular wave (d 0) was determined by ultrasonography. Starting on d 1, cows received LH (40 microg/h; n = 7) or saline (2 mL/h; n = 5) in hourly pulses for up to 5 (n = 5) or 7 (n = 7) d. On d 2, all cows received two injections of PGF2alpha, 12 h apart. In experiment 2, 14 lactating cows (7 to 12 d postpartum) received LH (40 microg/h; n = 7) or saline (1 mL/h; n = 7) in hourly pulses for 7 d, beginning 24 h after start of the first follicular wave. Daily samples were used to determine serum concentrations of progesterone (P4), estradiol-17beta (E2), LH, and FSH. Profiles of LH were determined from blood samples collected at 12-min intervals for 8 h on d 3. During infusion of LH, serum P4 and FSH were similar across treatments in both experiments. Serum E2 concentrations were similar in experiment 1, but serum E2 was greater on d 2, 3, and 5 in LH-treated cows in experiment 2. Infusion increased LH pulse frequency and amplitude in both experiments. Formation of cysts did not differ between LH- and saline-treated cows in either experiment (1 of 7 vs. 0 of 5 and 1 of 6 vs. 0 of 7, respectively). Cows that ovulated had similar intervals to ovulation in experiment 1 [6.0 +/- 0.1 d (LH) vs. 6.4 +/- 0.2 d (saline)], but in experiment 2, ovulation was 14 d earlier in LH-treated cows (5.6 +/- 1.8 d vs 19.9 +/- 1.5 d). In conclusion, high concentrations of LH are not solely responsible for formation of cysts in lactating dairy cows. Pulsatile infusion of LH stimulated follicular growth and steroidogenesis and decreased time to first ovulation in anestrous postpartum cows.

Animals↗

Growth hormone priming as an adjunct treatment in superovulatory protocols in the ewe alters follicle development but has no effect on ovulation rate.

This study investigated the effect of FSH alone and rGH priming followed by FSH treatment on follicle populations, follicular fluid concentrations of components of the IGF system and steroids, and the ovulation rate in sheep. Estrus was synchronized with progestagen sponges. Ewes (n = 10/group) in Group 1 served as untreated controls, while those in Groups 2 to 5 received a standard superovulatory treatment of 1.1 mg i.m. oFSH twice daily for 4 d. In addition, ewes in Groups 3 and 5 were administered rGH (15 mg/d, i.m.) for the 7 d prior to FSH treatment. Groups 1, 2 and 3 were sacrificed just prior to the LH surge; Groups 4 and 5 were allowed to ovulate. Daily plasma samples were collected to monitor GH, IGF-1 and insulin levels. All follicles > or = 1.0 mm from Groups 1, 2 and 3 were counted, and follicular fluid from follicles > or = 2.5 mm was assayed for estradiol, testosterone, IGF-1 and IGFBPs. Compared with the control, treatment with rGH + FSH but not FSH alone increased (P < 0.001) plasma concentrations of GH, IGF-1 and insulin. The mean number of large-(> or = 4.5 mm) and medium-sized (2.5 to 4.0 mm) follicles was increased (P < 0.01), and the mean number of small (< or = 2.0 mm) follicles was decreased (P < 0.001) by FSH treatment. The mean number of medium-sized (2.5 to 4.0 mm) follicles was further increased (P < 0.05) by rGH priming. Estradiol concentration in medium but not in large estrogenic follicles was increased (P < 0.05) by rGH priming, whereas testosterone concentration in estrogenic follicles was not altered. Components of the IGF system in medium-sized estrogenic follicles were similar in all treatment groups; however, in large estrogenic follicles rGH increased IGF-1 concentrations (P < 0.05) and intensity of the 44-42 kDa IGFBP band (P < 0.01). Priming with rGH did not alter superovulatory responses. These results show that rGH priming, when used as an adjunct to FSH treatment in ewes, alters components of the IGF system in large estrogenic follicles and increases the number and physiological maturity of medium-sized follicles in the ovary; it does not however alter ovulation rate responses.

Animals↗

Sexual behavior and pregnancy rate of Bos indicus bulls.

To study sexual behavior and pregnancy rate of Zebu bulls, 9 Brahman and 6 Nelore bulls were first examined for libido and serving capacity, semen characteristics and scrotal circumference, a week before being placed, individually, into a group of 16 estrus-syncronized heifers to observe mating activity. Scores for libido (0 to 10) and for serving capacity (based on the number of services per period) were assigned for each bull over a 15-min period. Estrus was synchronized with Norgestomet (3 mg, s.c. implant and 3 mg, i.m.) and estradiol valerate (5 mg, i.m.). The implants remained in place for 10 d, then 40 h after their removal a bull was placed with the heifers. Four Brahman and 6 Nelore bulls were monitored for 2 h while the remaining 5 Brahman bulls were observed for 30 h. Number of services and mounts per bull, the hour at which each event took place, and the identification of the heifers serviced were recorded. Pregnancy was diagnosed by transrectal palpation 50 to 60 d after mating. Mean libido score (+/- SEM) of the 15 bulls was 6.4 +/- 0.5, and mean serving capacity was 0.53 +/- 0.19; both of these measures were correlated with age (r = 0.78, P < 0.01, and r = 0.56, P < 0.05, respectively). In the 2-h trials, the number of mounts was correlated with the libido score (r = 0.73, P < 0.05), and the number of services was correlated with age (r = 0.76, P < 0.05) and with libido and serving capacity score (r = 0.91 and 0.92, P < 0.01). In the 30-h trials, the mean (+/- SEM) number of mounts and of services per bull was 74.4 +/- 21.1 and 14.6 +/- 0.5, respectively. The mount-to-service ratio and percentage of serving efficiency were 5.1 +/- 1.4, and 19.8 +/- 4%, respectively. Libido score was correlated with the mount-to-service ratio (r = -0.86, P < 0.10) and with serving efficiency (r = 0.84, P < 0.10). The percentage of heifers serviced, of those serviced 2 or more times and of those pregnant was 55 +/- 5.3; 47.1 +/- 6.4, and 59.2 +/- 5.8, respectively. Pregnancy rate was not correlated with any trait (P > 0.10). It was concluded that libido and the serving capacity test seem to indicate the mating potential of Zebu bulls over a relatively short period of time (e.g. 30 hours), in which resultant pregnancy rates are comparable to those of European bulls.

Aging↗

Mitochondrial aggregation patterns and activity in porcine oocytes and apoptosis in surrounding cumulus cells depends on the stage of pre-ovulatory maturation.

In this study, we evaluated the distribution and oxidative activity of mitochondria in ex vivo pre-ovulatory porcine oocytes using the fluorescence probe MitoTracker CMTM Ros Orange. Cumulus-oocyte complexes (COCs) were classified according to cumulus morphology and time from hCG administration. The meiotic configuration of the oocytes and the degree of apoptosis in the surrounding cumulus cells were also evaluated. Estrus was synchronized in 45 crossbred Landrace gilts by feeding altrenogest for 15 days and administering 1000 IU PMSG on Day 16. The LH peak was simulated by treatment with 500 IU hCG, given 80 h after PMSG. Endoscopic oocyte recovery was carried out 2 h before or 10, 22, or 34 h after hCG administration. Altogether 454 COCs were aspirated from follicles with a diameter of more than 5 mm. Cumulus morphology in the majority of COCs recovered 2 h before and 10 h after hCG was compact (60.4 and 52.7%, respectively; P<0.05). At 22 h after hCG, COC morphology changed significantly from 10 h dramatically: 74% of COCs had an expanded cumulus (P<0.01). At 34 h after hCG, 100% of recovered COCs had an expanded cumulus. The percentage of oocytes with a mature meiotic configuration differed among COC morphologies and increased as the interval after hCG administration increased (P<0.05). The type of mitochondrial distribution in the oocytes (n=336) changed from homogeneous to heterogeneous as the interval after hCG administration increased (P<0.01) and was associated with the cumulus morphology. Representative mitochondrial distributions were found as follows: -2 h: fine homogeneous in compact and dispersed COCs; 10 h: granulated homogeneous in compact and dispersed COCs; 22 h: granulated homogeneous in expanded COCs; and 34 h: granulated heterogeneous and clustered heterogeneous in expanded COCs (P<0.01). The oxidative activity of mitochondria measured by fluorescence intensity (Em: 570 nm) per oocyte after Mitotracker CMTM Ros Orange labeling increased in the oocyte as the post-hCG interval increased (P<0.01) and depended on the type of mitochondrial distribution. Lowest oxidative activity of mitochondria was found in oocytes with fine homogeneous distribution (253.1+/-9.4 microA). The oxidative activity increased (334.4+/-10.3 microA) in oocytes with granulated homogeneous distribution of mitochondria, and reached highest level in oocytes with granulated heterogeneous (400.9+/-13.0 microA) and clustered heterogeneous distributions (492.8+/-13.9 microA) (P<0.01). Mitochondrial activity in oocytes coincided with apoptosis in surrounding cumulus cells which increased in a time-dependent manner during pre-ovulatory maturation in vivo (P<0.01). These results indicate that there is a relationship between meiotic progression, cumulus expansion and mitochondrial redistribution and their oxidative activity during final pre-ovulatory maturation in pig oocytes. It appears that increased levels of mitochondrial activities in oocytes are correlated to increased levels of apoptosis in surrounding cumulus cells, in which mitochondria may play a role.

Animals↗

Effects of short- or long-term infusions of acetate or propionate on luteinizing hormone, insulin, and metabolite concentrations in beef heifers.

Two trials were conducted to evaluate the effects of short- (Trial 1) or long-term (Trial 2) intraruminal isocaloric infusions of acetate or propionate on secretion of LH, insulin, and selected metabolites in short- or long-term energy-restricted beef heifers. In Trial 1, 16 Angus heifers were assigned on d 6 to 12 of a synchronized estrous cycle (estrus = d 0) to a body weight-maintenance (BWM; n = 4) or an energy-restricted, body weight-loss (BWL; n = 12) treatment. On d 12 of a synchronized estrous cycle, heifers received PGF2alpha to synchronize estrus, and 12 h later BWL heifers received intraruminal, isocaloric infusions of acetate, propionate, or vehicle for 6 h and BWM heifers received vehicle concurrently. Mean plasma LH and LH pulse frequencies and amplitudes were not affected by treatment (P > .05). In contrast, infusion of propionate increased plasma insulin (P < .05) and reduced plasma concentration of NEFA (P < .05). In Trial 2, six ovariectomized Angus heifers were energy-restricted for 30 d. On d 14 and 26 of restriction, heifers began receiving intraruminal isocaloric infusions of acetate or propionate for 96 h in a switchback approach. Intraruminal infusions of vehicle for 6 h preceded infusions of acetate or propionate. Jugular blood was collected at 12-min intervals during infusions of vehicle and during the last 6 h of infusion of acetate or propionate. Mean concentration of LH and amplitude of pulses of LH were lower during acetate vs propionate or vehicle infusion (P < .05). Infusion of propionate increased insulin relative to acetate or vehicle infusion (P < .05). Plasma NEFA were reduced by infusion of propionate (P < .05) and increased by infusion of acetate (P < .05).

3-Hydroxybutyric Acid↗

Trypanosoma cruzi: the development of estrus cycle and parasitemia in female mice maintained with or without male pheromones.

Female BALB/c or C57B1/6 mice, kept in small groups of three or five animals with or without male odor, all had a similar progesterone and corticosterone level, mean number of estrus and duration of estrus cycle. However, if males were kept in the same room, the mean duration of the estrus cycle was longer for both strains; and C57B1/6 females had a significantly higher number of estrus than BALB/c mice and showed a tendency to synchronize the estrus cycle within a group. After infection of females of both mouse strains with vector-derived metacyclic trypomastigotes of Trypanosoma cruzi, anestrus with intense phlegm production occurred during the acute phase of infection and this was positively correlated with higher parasitemia. Within individual groups of BALB/c mice, the female with the relatively highest corticosterone and progesterone level had the lowest parasitemia. In groups kept separate from male pheromones, one or two females in each group developed high parasitemias.

Anestrus↗

Caprine luteinizing hormone isoforms during the follicular phase and anestrus.

The relative proportion of the circulating luteinizing hormone isoforms in goats during follicular phase (pre-ovulatory peak; F) and anestrus (A) was investigated. Estrus was synchronized in six goats with a prostaglandin analogue. After estrus was detected, blood samples were taken at 1 h intervals for 24 h. Four anestrous goats received 100 microg i.v. of GnRH and blood samples were collected every 15 min for 5 h. Samples with the greatest LH concentration in follicular phase and after GnRH administration (anestrus) were analyzed by chromatofocusing and eluted with a pH gradient from 10.5 to 3.5. For quantification purposes eluted LH was grouped into basic (pH> or =7.5), neutral (pH 7.4-6.5) and acidic isoforms (pH< or =6.4) as well as by pH unit. In both physiological conditions (PC), basic and acidic isoforms were greater than the neutral. With this grouping criteria, there was an interaction between PC and pH group, with the proportion of neutral isoforms being greater (p<0.05) in A (12.0+/-0.8%) as compared with F (5+/-2%). Analysis by pH unit showed a very basic group of eluted isoforms (pH> or =10), which amounted to a percentage of 6.0+/-0.4% of the total observed during A, and 3+/-1% during F (p<0.05). Predominant isoforms in A eluted in the pH range 9.99-9.0 (42+/-3%) as compared to 7+/-3% (p<0.01) in that pH range in F. In contrast, the predominant isoforms in F eluted in the pH range 8.99-8.0, representing 55+/-8%, while in A the proportion was 11+/-2% (p<0.01). Isoforms eluted at the pH range 7.9-7 represented a significantly greater proportion during A (5.0+/-0.6%) as compared with F (3+/-1%). This is the first report on goat LH circulating isoforms. During A the LH isoforms secreted by the pituitary are more basic than during F.

Anestrus↗

Estrous behavior and endocrine status of dairy heifers with varied intakes of phosphorus.

We investigated whether dietary phosphorus affected estrous intensity of heifers. A total of 48 heifers (Holstein and Jersey) were allotted to diets containing approximately 73% (low), 138% (medium), or 246% (high) of the phosphorus recommended by the National Research Council. Heifers were observed continuously for 48 h beginning 36 h after an estrous-synchronizing treatment of prostaglandin F2 alpha. Heifers began standing estrus 55 +/- 12.6 h (standard deviation) after prostaglandin, with onset randomly throughout the day. Standing estrus lasted 15.4 +/- 4.1 h (standard deviation), and heifers stood to be mounted 3.68 +/- 2.38 (standard deviation) times per hour. Dietary phosphorus concentration altered inorganic phosphorus of blood serum but did not affect significantly overall estrous behavior or blood serum concentrations of progesterone, estradiol, or luteinizing hormone measured around the time of estrus. We conclude that a dietary phosphorus inadequacy as in this study does not alter reproductive endocrine function or estrous intensity in heifers.

Animals↗

Utilization of the growth phase of the first follicular wave for bovine oocyte collection improves blastocyst production.

Characteristics of the follicle population and oocyte developmental competence at selected stages of follicular development were studied in cows with the aim to increase embryo production derived from oocytes collected by transvaginal aspiration. In Experiment 1, the growth phase before dominant follicle selection and the low dominant phase during dominant follicle regression were compared. Twenty-four cyclic Holstein cows, 4 to 6 yr of age, were divided into 2 groups. Animals were synchronized using two injections of prostaglandin F2alpha at 11 d intervals, and onset of estrus was determined (Day 0). Using ultrasonography, all follicles were counted and classified. Oocytes were aspirated once on Days I through 3 (Group 1, n=5) or Days 15 and 16 (Group 2, n=3) of the estrus cycle. The experiment was carried out in 3 replicates. In Experiment 2, the growth phase of the first follicular wave before dominant follicle selection was characterized in detail. Twelve cows of the same breed and age were divided into 3 groups. Their first estrus was synchronized as in Experiment 1, and each following estrus was induced using one injection of prostaglandin F2alpha administered 4 to 6 d after each aspiration performed. The ovaries were examined, and oocytes were collected repeatedly (total of 5 times per cow) on Days 1 (Group 3, n=4), 2 (Group 4, n=4) or 3 (Group 5, n=4) after estrus at 10 d intervals during a 40 d period. Viable oocytes were matured, fertilized and cultured using the standard methods. In Experiment 1, the mean numbers (+/-SD) of all follicles and of recovered and viable oocytes per donor were higher in Group 1 than in Group 2, but only the mean numbers (+/-SD) of larger follicles and recovered oocytes were statistically significant (8.0 +/- 0.6 and 6.2 +/- 0.6.vs. 3.3 +/- 0.5 and 2.8 +/- 0.2; P< 0.05). In Experiment 2, the percentage of larger follicles out of all visible follicles and the mean numbers (+/-SD) of larger follicles per donor were significantly higher (P<0.05) in Groups 4 (75.7 and 9.1 +/- 2.7) and 5 (66.3 and 8.5 +/- 2.9) when compared to Group 3 (27.9 and 3.8 +/- 0.8). The development rate of fertilized oocytes was significantly higher (P<0.05) in Groups 4 (27.8) and 5 (27.5) than in Group 3 (12.8). It can be concluded that it is possible to improve the efficiency of transvaginal aspiration and in vitro embryo production by utilization of the growth phase of the first follicular wave before dominant follicle selection.

Animals↗

Chronic utero-ovarian vein catheterization with subsequent occlusion prolongs the estrous cycle and changes electromyographic activity in the myometrium of ewes.

The objective of our study was to determine the effect of chronic utero-ovarian vein catheterization in ewes on estrous cycle length, plasma progesterone (P) concentration, and myometrial electromyographic activity. Cyclic ewes with inferior vena cava catheters were used as controls. Estrus was synchronized in ten ewes and 10 to 12 d following estrus, the ewes were anesthetized, fitted with myometrial electromyograph leads and with utero-ovarian vein (n = 5) or inferior vena cava (n = 5) catheters. After surgery, ewes returned to estrus as expected (16 to 18 d interestrus interval). The second cycle of four of five ewes with utero-ovarian vein catheters were prolonged (40 to 58 d). The inferior vena cava catheterized ewes had normal length second cycles. Plasma P concentrations reflected the estrous cycles: low (</= 0.01 ng/ml) at estrus and 2 to 3 ng/ml at midcycle. The inferior vena cava catheterized ewes had decreased plasma P concentrations after luteolysis (Days 13 to 14) while four of five utero-ovarian vein catheterized ewes maintained elevated plasma P concentrations for 20 to 58 d. Catheterization affected the myometrial electromyograph; short events (16 to 180 sec) were increased on Days 5 to 13 in utero-ovarian vein as compared with inferior vena cava catheterized ewes (P < 0.05); long events (180 to 900 sec) tended to decrease from Days 1 to 15 in utero-ovarian vein ewes, but this was not statistically significant (P > 0.05).

Journal Article↗

Effect of timing of artificial insemination on gender ratio in beef cattle.

It was recently reported that cows inseminated at approximately 10 or 20 h before an expected ovulation deliver predominately a bull or heifer calf, respectively. The objective of this study was to further investigate the effect of timing of insemination on the gender of offspring in cattle. Angus heifers (n = 41) and cows (n = 98) were used in the study. Heifers were synchronized with a 16-d treatment of melengestrol acetate followed 17 d later with an injection of PGF2alpha. Cows were synchronized with GnRH followed 7 d later with PGF2alpha. A HeatWatch electronic estrus detection system was used to determine the onset of estrus. Based on previous studies, it was assumed that ovulation occurs approximately 32 h after the onset of estrus. Therefore, animals were artificially inseminated at either 8 to 10 h (early; > or = 20 h before expected ovulation) or 20 to 25 h (late; < or = 10 h before expected ovulation) after the onset of estrus. Sixty to 80 d after insemination, ultrasonography was used to confirm pregnancy status and to determine the gender of fetuses. Gender of calves was subsequently confirmed at calving. Data were analyzed for effects of time of insemination and sire or semen batch on gender ratio, as well as any effect of length and/or intensity of estrus on conception rate and gender ratio. Twenty-nine of 41 heifers and 69 of 98 cows were detected in estrus after synchronization and were inseminated; 20 of 29 heifers and 48 of 69 cows were subsequently confirmed pregnant. Neither the length of estrus nor its intensity (number of mounts) had an effect on pregnancy rate or gender ratio (P > or = 0.418). Timing of insemination (early versus late) had no effect on gender ratio (P = 0.887). Semen from 13 sires representing 17 lots was used to inseminate the cows and heifers. No differences (P = 0.494) were detected in the gender ratios resulting from different sires or semen batches. In contrast to previous findings, our results indicate that inseminating beef cattle at approximately 20 or 10 h before an expected ovulation does not alter the gender ratio of the resultant calves.

Animals↗