Ovulation in the tree shrew (Tupaia belangeri) induced by gonadotrophins

The induction of ovulation by exogenous gonadotrophins is an important approach for recovering oocytes used for studies on the reproductive biology of some mammals. In the present study, pregnant mare serum gonadotrophin (PMSG) and human chorionic gonadot

Unpredictable Cases of Complicated Ovarian Hyperstimulation in IVF

Objective-To characterize a subpopulation of complicated cases of ovarian hyperstimulation syndrome (OHSS). Method-Descriptive retrospective study. Results-0.75% of our IVF-ET population suffered from OHSS. Among this group, 33% did not exhibit any recognized risk criteria of OHSS in terms of infertility characteristics and ovarian response to exogenous gonadotrophins. Only severe (ascites) OHSS cases were considered (n = 5) in this study. Previous IVF-ET attempts had been uneventful and during the complicated trial, estradiol peak levels and numbers of oocytes retrieved remained below 2,500 pg/mL (conversion factor to SI unit, 3.671) and 10, respectively. In all cases, the luteal phase was supplemented by hCG and all patients became pregnant. A threshold level of exogenous and/or endogenous hCG seems to be responsible for the occurrence of OHSS. Conclusion-One-third of the patients developing an ovarian hyperstimulation syndrome after IVF-ET had not previously shown risk criteria. A causal role of exogenous and/or endogenous hCG is suggested.

Potential biomarkers on gametogenesis process and in sperm quality of european EEL (Anguilla anguilla)

Dissertação de mestrado, Aquacultura, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Strategies to improve fertility in Bos indicus postpubertal heifers and nonlactating cows submitted to fixed-time artificial insemination

Two experiments were designed to evaluate strategies to increase fertility of Bos indicus postpubertal heifers and nonlactating cows submitted to a fixed-time artificial insemination (TAI) protocol consisting of an intravaginal device containing 1.9 g of progesterone (CIDR) insertion + estradiol benzoate on Day 0, CIDR withdrawal + estradiol cypionate on Day 9, and TAI on Day 11. In Experiment 1, heifers (n = 1153) received a new or an 18-d previously used CIDR and, on Day 9, prostaglandin F(2 alpha) (PGF(2 alpha)) + 0, 200, or 300 IU equine chorionic gonadotropin (eCG). Heifers treated with a new CIDR had greater (least squares means +/- SEM) serum concentration of progesterone on Day 9 (3.06 +/- 0.09 ng/mL vs. 2.53 +/- 0.09 ng/mL; P < 0.05) and a smaller follicle at TAI (11.61 +/- 0.11 nim vs. 12.05 +/- 0.12 mm; P < 0.05). Heifers with smaller follicles at TAI had lesser serum progesterone, concentrations on Day 18 and reduced rates of ovulation, conception, and pregnancy (P < 0.05). Treatment with eCG improved (P < 0.05) follicle diameter at TAI (11.50 +/- 0.10 mm, 11.90 +/- 0.11 mm, and 12.00 +/- 0.10 mm, for 0, 100, and 200 IU, respectively), serum progesterone concentration on Day 18 (2.77 +/- 0.11 ng/mL, 3.81 +/- 0.11 ng/mL, and 4.87 +/- 0.11 ng/mL), and rates of ovulation (83.8%, 88.5%, and 94.3%) and pregnancy (41.3%, 47.0%, and 46.7%). In Experiment 2, nonlactating Nelore cows (n = 702) received PGF(2 alpha) treatment on Days 7 or 9 and, on Day 9, 0 or 300 IU cCG. Cows receiving PGF(2 alpha) on Day 7 had lesser serum progesterone concentrations on Day 9 (3.05 +/- 0.21 ng/mL vs. 4.58 +/- 0.21 ng/mL; P < 0.05), a larger follicle at TAI (11.54 +/- 0.21 mm vs. 10.84 +/- 0.21 mm; P < 0.05), and improved (P < 0.05) rates of ovulation (85.4% vs. 77.0%), conception (60.9% vs. 47.2%), and pregnancy (52.0% vs. 36.4%). Treatment with eCG improved (P < 0.05) serum progesterone concentration on Day 18 (3.24 +/- 0.14 ng/mL vs. 4.55 +/- 0.14 ng/mL) and the rates of ovulation (72.4% vs. 90.0%) and pregnancy (37.5% vs. 50.8%). In conclusion, giving PGF(2 alpha) earlier in the protocol in nonlactating cows and eCG treatment in postpubertal heifers and nonlactating cows improved fertility in response to a TAI (progesterone + estradiol) protocol. (C) 2009 Elsevier B.V. All rights reserved.

Progesterone-based estrous synchronization protocols in non-suckled and suckled primiparous Bos indicus beef cows

In Experiment 1, effects of an intravaginal progesterone insert (CIDR) for synchronization of estrus in non-suckled cycling primiparous Bos indicus beef cows were evaluated. Cows received either no treatment (Control), prostaglandin F(2 alpha) (PGF), or CIDR (6 d)+prostaglandin F(2 alpha) (CIDR+PGF). There were greater (P<0.05) percentages of cows inseminated and pregnant in 4 d of breeding season in CIDR+PGF (81.6%; 48.7%) than in PGF (53.8%; 32.5%) and Control cows (14.6%; 8.5%). In Experiment 2, efficacy of a treatment with CIDR (6 d)+prostaglandin F(2 alpha)+48 h temporary weaning (CIDR+PGF+TW) associated or not with equine chorionic gonadotropin (eCG) for synchronization of estrus in suckled primiparous B. indicus beef cows was evaluated. Cows were assigned to receive either no treatment (Control), CIDR+PGF+TW, CIDR+PGF+TW+200 IU of eCG, or CIDR+PGF+IW+400 IU of eCG. There were greater (P<0.05) percentages of insemination and pregnancy in 4 d of breeding season in treatment groups (47.0%; 27.5%) than in Control cows(4.5%; 3.0%) and in cows treated with eCG (50.9%; 29.4%) than in cows treated only with CIDR+PGF+TW (39.4%: 23.7%). No effects of eCG dosage were detected. The percentage of cows inseminated and pregnant in a 30 d breeding season were greater (P<0.05) in cows assigned to synchronization treatments (53.3%; 37.2%) than in Control cows (35.3%; 21.4%). In the beginning of the BS, non-suckled primiparous cows treated with CIDR+PGF with estrous cycles having been initiated following calving had greater estrous detection and pregnancy rates than PGF and Control cows, and suckled primiparous cows had improved estrous detection and pregnancy rates when eCG was associated with CIDR+PGF+TW. (C) 2009 Elsevier B.V. All rights reserved.

Fixed-time artificial insemination with estradiol and progesterone for Bos indicus cows II: Strategies and factors affecting fertility

In Experiments 1, 2, and 3, we evaluated the effects of temporary weaning (TW), equine chorionic gonadotropin (eCG), and follicle-stimulating hormone (FSH) treatments on results of it fixed-time artificial insemination (TAI) protocol in postpartum Bos indicus cows. In Experiment 1, treatment with 400 IU eCG or with TW for 48 h consistently improved pregnancy rates (PRs) at TAI, but, in Experiment 2, FSH treatment was less effective than eCG or TW. In Experiment 3, the inclusion of eCG treatment in cows subjected to TW did not improve PRs. We concluded that TW or 400 IU eCG should be included in the TAI protocol in postpartum Bos indicus cows to enhance fertility. In Experiment 4, we used records front heifers and cows treated with the proposed protocol during the 2006-2007 (n = 27,195) and 2007-2008 (n = 36,838) breeding seasons from multiple locations in Brazil to evaluate factors potentially affecting PRs. Overall PR at TAI was 49.6% (31,786 of 64,033). Pregnancy rate differed (P < 0.01) among farm within location (results ranging between 26.8% and 68.0%; P < 0.01), cow group within farm, by breed (Bos indicus, 48.3% [26,123 of 54,145]; Bos taurus, 61.7% [3652 of 5922]; and crossbred Bos indicus x Bos taurus, 50.7% [2011 of 3966]), category (nulliparous, 39.6% [2095 of 52901] suckled primiparous, 45.2% [3924 of 8677]; suckled multiparous, 51.8% [24,245 of 46,767]; and nonsuckled multiparous, 46.1% [1522 of 3299]), body condition score at TAI (<= 2.5 43.0% [3409 of 7923]; 3.0, 49.6% [18,958 of 38,229]; and >3.5, 52.7% [9419 of 17,881]). Days postpartum at beginning, of protocol did not affect PR (30 to 60 d, 47.6% [4228 of 8881]; 61 to 90 d, 51.7% 116,325 to 31,5721; and 91 to 150 d, 50.8% [7616 to 14,991]; P > 0.1). Pregnancy rate was also consistently affected (P < 0.01) by sire (results ranging from 7.2% to 77.3%) and artificial insemination technician (results ranging from 15.1% to 81.8%). (C) 2009 Elsevier B.V. All rights reserved.

Time-dependent effects of lead on rat reproductive functions

The effects of exposure to lead on endocrine function and the reproductive parameters were studied in pubertal rats treated with 1.0 g l(-1) lead acetate in drinking water for 20 days (subacute group) or 9 months (chronic group) in addition to i.v. injections of lead acetate (0.1 mg 100 g(-1) body wt.) every 10 (subacute group) or 15 days (chronic group). Although basal levels of testosterone were higher both in plasma and in testes of acutely intoxicated animals, the circulating levels of luteinizing hormone (LH) were not affected in either group, nor was the LH-releasing hormone content of the median eminence. The density of [I-125]LH/human chorionic gonadotrophin (hCG) binding sites in testicular homogenates was reduced by saturnism in both groups, concomitant with a significantly increased apparent affinity constant of the hormone-receptor complex. These data can be viewed as the result of a mixture of specific lead toxicity (e.g. at the enzyme level) with other more general actions (e.g. at the level of the hypothalamus-pituitary-testicular axis).

Do high progesterone concentrations decrease pregnancy rates in embryo recipients synchronized with PGF2 alpha and eCG?

The objective of this study was to evaluate the effects of equine chorionic gonadotropin (eCG) treatment on the number of induced accessory corpora lutea (CL), plasma progesterone concentrations and pregnancy rate in cross-bred heifers after transfer of frozen-thawed (1.5 M ethylene glycol) embryos. All recipients received 500 mug PGF2alpha (dl-cloprostenol, i.m.) at random stages of the estrous cycle (Day 0) and were observed for estrus for 7 days. on Day 14, heifers detected in estrus between 2 and 7 days after PGF2alpha treatment were randomly allocated to four groups (n = 83 per group) and given 0 (control), 200, 400, or 600 IU of eCG. Two days later (Day 16), these recipients were given PGF2a and observed for estrus. Six to eight days after detection of estrus, plasma samples were collected to determine progesterone concentration and ultrasonography was performed to observe ovarian structures. Heifers with multiple CL or a single CL >15 mm in diameter received an embryo by direct transfer. Embryos of excellent and good quality were thawed and transferred to the recipients by the same veterinarian. Pregnancy was diagnosed by ultrasonography and confirmed by transrectal palpation 21 and 83 days after embryo transfer (ET), respectively. Plasma progesterone concentrations on the day of transfer (Day 7 of the estrous cycle) were 3.9 +/- 0.7, 4.2 +/- 0.4, 6.0 +/- 0.4, and 7.8 +/- 0.6 ng/ml for groups Control, 200, 400, and 600, respectively (Control versus treated groups P = 0.009; 200 versus 400 and 600 groups P = 0.0001; and 400 versus 600 P = 0.012). Conception rates 83 days after ET were 41.9, 50.0, 25.0, and 20.9% for groups Control, 200, 400, and 600, respectively (200 versus 400 and 600 groups P = 0.0036). In conclusion, an increase in progesterone concentration, induced by eCG treatment, did not improve pregnancy rates in ET recipients. Conversely, there was a decline in conception rates in the animals with the highest plasma progesterone concentrations. (C) 2003 Elsevier B.V. All rights reserved.

Ovarian follicle diameter at timed insemination and estrous response influence likelihood of ovulation and pregnancy after estrous synchronization with progesterone or progestin-based protocols in suckled Bos indicus cows

The objectives of the present study were to evaluate factors associated with estrous synchronization responses and pregnancy per insemination (P/AI) in Bos indicus beef cows submitted to progesterone-based fixed-time artificial insemination (FTAI) protocols. A total of 2388 cows (1869 Nellore and 519 crossbred Nellore x Angus) from 10 commercial farms were evaluated to determine the relationships among breed, body condition score (BCS) on the first day of the FTAI protocol, the occurrence of estrus between progesterone device removal and FTAI and diameter of largest ovarian follicle (LF) at FTAI on estrous synchronization responses and P/AI. Cows (n=412 primiparous; 1976 multiparous) received an intravaginal device containing progesterone or an ear implant containing norgestomet (a progestin), and an injection of estradiol at the beginning of the estrous synchronization protocol. Body condition was scored using a 1-5 scale on the first day of the FTAI protocol and at 30-60 days postpartum. Females received 300IU of equine chorionic gonadotropin (eCG) and PGF(2 alpha) on the day the progesterone device/implant was removed and were inseminated 48-60h later. At insemination, cows (n=2388) were submitted to an ultrasonographic exam to determine the diameter of the LF. Follicles were classified into four categories based on mean and standard deviation (SD) of the LF (LF1 = two SD below the mean; LF2 = mean minus one SD; LF3 = mean plus one SD; LF4 = two SD above the mean). Ovulation rate was determined in a subset of cows (n=813) by three consecutive ultrasonographic exams: (1) at time of progesterone device/implant removal, (2) at time of FTAI and (3) 48 h after FTAI. Ovulation was defined as the disappearance of a large follicle (>= 8.0 mm) that was previously recorded. Estrus was determined in a subset of the cows (n = 445) by the activation of a detection of estrous patch placed on the tail head on the day of progesterone device/implant removal. Pregnancy was diagnosed 30 days after FTAI. Pregnancy was influenced (P = 0.001) by follicle diameter [LF1 = 27.5% (81/295), LF2 = 46.6% (328/705), LF3 = 57.9% (647/1118), LF4 = 63.3% (171/270)] and the occurrence of estrus [estrus = 67.7% (174/257) and no estrus = 36.2% (68/188)]. Follicle diameter at FTAI influenced ovulation rate [LF1 = 42.5% (34/80), LF2 = 73.9% (161/218), LF3 = 95.8% (407/425), LF4 = 97.8% (88/90)], the occurrence of estrus [LF1 = 54.8% (51/93), LF2 = 33.6% (43/128), LF3 = 68.9% (126/183), LF4 = 90.2% (37/41)] and P/AI among cows that had ovulations [LF1 =32.4% (11/34), LF2 = 50.3% (81/161), LF3 = 60.0% (244/407), LF4 = 68.2% (60/88)]. Improving estrous responses between progesterone device withdrawal and FTAI and increasing the diameter of the LF at FTAI may be important aspects to achieve improved estrous synchronization responses and P/AI following progesterone/progestin and estradiol based FTAI protocols in suckled Bos indicus cows. (C) 2010 Elsevier B.V. All rights reserved.

Effect of capacitation of stallion sperm with polyvinylalcohol or bovine serum albumin on penetration of bovine zona-free or partially zona-removed equine oocytes

Experiments were conducted to study effects of macromolecules on stallion sperm capacitation and fertilization as determined by penetration of bovine zona-free and equine partially zona-removed oocytes. Stallion sperm were capacitated in TYH medium (modified Krebs-Ringer bicarbonate) supplemented with either 1 mg/mL of polyvinylalcohol (PVA) or 4 mg/ mL of BSA. Capacitation was induced with 8 bromoadenosine cyclic monophosphate (8BrcAMP; 0.5 mM) alone or in combination with 0.1 μM of ionomycin. Intraspecies gametes were co-incubated in TYH/PVA or TYH/ BSA for 18 to 20 h. For zona-free bovine oocytes, penetration rate (35%) with the combination of 8BrcAMP and ionomycin in PVA-containing medium was higher (P < 0.05) than any treatment in BSA-containing medium (5 to 6%). A similar study was conducted using equine oocytes with partially removed zonae. Sperm capacitated and used for in vitro fertilization (IVF) in PVA-containing medium had higher penetration rates (P < 0.01) than sperm in BSA-containing medium (54 vs. 11%). The effect of equine preovulatory follicular fluid on bovine oocyte penetration was assessed. Bovine oocytes were matured in tissue culture medium-199 with 0, 20, 50, or 100% equine preovulatory follicular fluid, and 1 IU/mL of equine chorionic gonadotropin. Stallion sperm were treated with 8BrcAMP + ionomycin in PVA- or BSA-containing media. The penetration rates of bovine zona-free oocytes by stallion sperm were again higher with PVA (47%) than BSA (18%; P < 0.01). Penetration rates of oocytes matured in 100% follicular fluid were higher (P < 0.05) than for oocytes matured with 0% follicular fluid. The effects of equine follicular fluid and PVA/BSA during sperm capacitation on standard bovine IVF were examined. Culture of bovine oocytes with equine follicular fluid did not affect oocyte maturation or penetration rates after IVF. Bovine sperm capacitated with heparin in PVA-containing medium yielded lower (P < 0.05) fertilization rates than those capacitated in BSA-containing medium when incubated with both zona-intact and zona-free bovine oocytes. In summary, PVA was superior to BSA for ionophore-induced capacitation of equine sperm for penetration of zona-free bovine oocytes or partially zona-removed equine oocytes, but not for standard bovine IVF with bovine sperm. Zona-free bovine oocytes may be useful for assaying in vitro capacitation and fertilization of stallion sperm. © 2003 American Society of Animal Science. All rights reserved.

Neither plasma progesterone concentrations nor exogenous eCG affects rates of ovulation or pregnancy in fixed-time artificial insemination (FTAI) protocols for puberal Nellore heifers

The objective was to evaluate the effects of plasma progesterone (P4) concentrations and exogenous eCG on ovulation and pregnancy rates of pubertal Nellore heifers in fixed-time artificial insemination (FTAI) protocols. In Experiment 1 (Exp. 1), on Day 0 (7 d after ovulation), heifers (n = 15) were given 2 mg of estradiol benzoate (EB) im and randomly allocated to receive: an intravaginal progesterone-releasing device containing 0.558 g of P4 (group 0.5G, n = 4); an intravaginal device containing 1 g of P4 (group 1G, n = 4); 0.558 g of P4 and PGF2α (PGF; 150 μg d-cloprostenol, group 0.5G/PGF, n = 4); or 1 g of P4 and PGF (group 1G/PGF, n = 3). On Day 8, PGF was given to all heifers and intravaginal devices removed; 24 h later (Day 9), all heifers were given 1 mg EB im. In Exp. 2, pubertal Nellore heifers (n = 292) were treated as in Exp. 1, with FTAI on Day 10 (30 to 36 h after EB). In Exp. 3, pubertal heifers (n = 459) received the treatments described for groups 0.5G/PGF and 1G/PGF and were also given 300 IU of eCG im (groups 0.5G/PGF/eCG and 1G/PGF/eCG) at device removal (Day 8). In Exp. 1, plasma P4 concentrations were significantly higher in heifers that received 1.0 vs 0.588 g P4, and were significantly lower in heifers that received PGF on Day 0. In Exp. 2 and 3, there were no significant differences among groups in rates of ovulation (65-77%) or pregnancy (Exp. 2: 26-33%; Exp. 3: 39-43%). In Exp. 3, diameter of the dominant ovarian follicle on Day 9 was larger in heifers given 0.558 g vs 1.0 g P4 (10.3 ± 0.2 vs 9.3 ± 0.2 mm; P < 0.01). In conclusion, lesser amounts of P4 in the intravaginal device or PGF on Day 0 decreased plasma P4 from Days 1 to 8 and increased diameter of the dominant follicle on Day 9. However, neither of these nor 300 IU of eCG on Day 8 significantly increased rates of ovulation or pregnancy. © 2011.

Progesterone-based strategies to induce ovulation in prepubertal Nellore heifers

Four experiments were conducted to evaluate hormonal strategies to induce ovulation in Nellore heifers. In experiment 1, heifers (N = 1039) received a controlled internal drug release (CIDR) of fourth use (CIDR-4) on Day -12 or no CIDR (CIDR-0). The CIDR was removed on Day 0 in the CIDR-4 treatment, and estrus detection and AI were performed from Days 1 to 7. On Day 8, heifers not detected in estrus were evaluated for CL presence and received the same treatment again, followed by estrus detection and AI from Days 21 to 27. All heifers in experiments 2 (N = 896), 3 (N = 839), and 4 (N = 948) received the CIDR-4 treatment on Day -12. In experiment 2, heifers were randomly assigned to a control group (no additional treatment) or to receive equine chorionic gonadotropin (eCG; 200 IU eCG im) on Day 0. In experiment 3, heifers received the same treatments as in experiment 2, or a treatment that included eCG and estradiol cypionate (ECP) (eCG+ECP; 200 IU im eCG plus 0.5 mg ECP im) on Day 0. In experiment 4, heifers received the treatments described in experiment 3 or only ECP (0.5 mg) on Day 0. In experiments 2 and 3, estrus detection and AI was performed from Days 1 to 7 and on Day 8, heifers not detected in estrus were evaluated for CL presence. In experiment 4, heifers were evaluated for presence of a CL between Days 10 and 14. In experiment 1 heifers treated with CIDR-4 had greater estrus detection, ovulation induction, and pregnancy rates than in the CIDR-0 group. In experiment 2, heifers treated with eCG had greater estrus detection, ovulation induction, and pregnancy rates in 7 days than heifers in the control group. In experiment 3, heifers treated with eCG+ECP had greater estrus detection, ovulation induction, and pregnancy rates than the control and eCG treatments. In experiment 4, ovulation induction was greater for heifers treated with eCG and eCG+ECP relative to control, but did not differ from the ECP treatment. In conclusion, the use of a CIDR of fourth use for 12 days and the addition of eCG and/or ECP at CIDR removal efficiently induced ovulation and increased pregnancy rates in prepubertal Nellore heifers. © 2013 Elsevier Inc.

Estrus synchronization and fixed time artificial insemination (FTAI) in dairy buffaloes during seasonal anestrus

The aim of this work was to study estrus synchronization and fixed time artificial insemination (FTAI) in dairy buffaloes during season anestrus. One hundred thirty-nine dairy buffaloes in seasonal anestrus were divided in two groups as G1(n=66) and G2(n=73). The protocols for both the groups were the same until day (D)14:D0 administration of 2.0 mg estradiol benzoate and implantation of progesterone device (P4) for 14 days; D14 removal of P4 plus 150 mg of cloprostenol and 400 IU of equine chorionic gonadotropin. On D16, G1 received 10 mg of buserelin and G2 100 mg deslorelin acetate. On D17, both the groups were submitted to FTAI. Ultrasonographic examinations of ovaries were performed on D0, D14, D16 and D17. Results showed that pregnancy rates in G1 and G2 were 20 and 41% (p<0.05) and the ovulation rates were 16.6 and 37%, respectively (p<0.05). The dominant follicle (DF) diameter on D16 was 7.9 mm in G1 and 8.9 mm in G2 (p>0.05). Thirty-five percent of the animals in G1 and 54.1% in G2 showed a diameter DF greater than 8.0 mm on D16 (p>0.05). Thus, it could be concluded that the protocols synchronized the estrus, leading the concentration of the parturitions in the period of low milk production. Deslorelin was more efficient than buserelin due the higher percentage of DF ovulation and higher pregnancy rates.

Suplementação do meio de maturação com gonadotrofinas placentárias na produção in vitro de blastocistos bovinos

Pós-graduação em Ciências Biológicas (Farmacologia) - IBB

Identificação precoce de neoplasia trofoblástica pós-molar pela curva de regressão normal da gonadotrofina coriônica humana

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB