Evaluation of von willebrand factor during pregnancy, lactation and oestrous cycle in bitches affected and unaffected by von willebrand disease

Plasmatic concentrations of von Willebrand Factor (vWF) increase during pregnancy in humans and dogs; however the mechanism of such increase is still not well defined. The aims of this study were: (i) to evaluate changes in vWF concentration during pregnancy and during the subsequent oestrous cycle in bitches affected and unaffected by von Willebrand Disease (vWD); (ii) to correlate the vWF levels and cortisol levels in both groups. Seven vWD affected (GI) and nine unaffected (GII) bitches were used. The animals were assessed during pregnancy, parturition, lactation and non-gestational oestrous cycle in 11 moments (Pregnancy 1, Pregnancy 2, Parturition, Lactation 1, Lactation 2, Lactation 3, Anestrus, Proestrus, Oestrus, Diestrus 1, and Diestrus 2). The following tests were performed; measurement of von Willebrand factor antigen (vWF:Ag), albumin and cortisol. In both groups, vWF concentration remained stable during the non-gestational oestrous cycle, but increased during pregnancy, with the highest value observed at parturition. Increases of 70% and 124% in vWF were seen in GI and GII, respectively, compared to anestrus. No correlation was found between vWF and cortisol. Values of vWF:Ag changed during pregnancy, with a peak at parturition, both in vWD affected and unaffected animals. Values of vWF were not altered in the different phases of the oestrous cycle following pregnancy in both groups. Evaluation of vWF during pregnancy can cause false negative results for vWD, but assessment can be performed at any point in the oestrous cycle of non-pregnant bitches. © 2012 Blackwell Verlag GmbH.

Maternidade para mães trabalhadoras: depressão pós-parto, stress, rede de apoio e conjugalidade

Pós-graduação em Psicologia do Desenvolvimento e Aprendizagem - FC

Gestational Trophoblastic Neoplasia Following Molar Ectopic Pregnancy A Case Report

BACKGROUND: Ectopic molar pregnancy is a gestational trophoblastic disease (GTD) of rare occurrence and therefore not always remembered as a diagnostic possibility.CASE: We describe a case of molar ectopic pregnancy in a primiparous woman who developed gestational trophoblastic neoplasia and required chemotherapy to achieve remission.CONCLUSION This case stresses the important role of histopathologic examination in establishing a diagnosis of ectopic molar pregnancy. Moreover, close follow-up of human chorionic gonadotropin levels is required when a GTD is suspected. (J Reprod Med 2008;53:579-582)

Fertility Rates Following Fixed-Time Artificial Insemination in Dairy Heifers in a Practical Progesterone-Based Protocol

Background: In bovines, more efficient management practices are important for maximizing profitability. In order to increase the pregnancy rates in artificial insemination (AI) programs, several hormonal protocols were developed to synchronize the follicular wave and the moment of ovulation in beef and dairy cattle. In dairy cattle, detection of estrus can be difficult due to a number of factors including the incidence of silent estrus. Hormonal treatments designed to control both luteal and follicular function has permitting efficient synchronizations of time of ovulation. Thus, the AI can be performed in a large number of animals on a fixed schedule without the need for detection of estrus. Using these management techniques, the fixed-time artificial insemination (TAI) can overcome the problem of accurate estrus detection and help in reducing the incidence of repeat breeding. In addition, with TAI in cattle operations, it is possible to facilitate management practices and commercialization, and to reduce the time and semen wasting with animals inseminated at incorrect times. The investigation of practical and efficient TAI protocols is important for reducing the labor and animal handling of TAI in dairy cattle, as well as for increasing the profitability of the cattle management system. This study was carried out in order to investigate the effectiveness of TAI in dairy heifers treated with a practical progesterone-based protocol.Materials, Methods & Results: This experiment was conducted at the university farm located in southwestern Brazil, during May 2009. Thirty-nine cycling crossbred dairy heifers were employed in this study. All animals received a single intramuscular injection of estradiol benzoate and intravaginal progesterone releasing device in a random stage of the estrous cycle (Day 0). on day 7 the animals were treated with PGF2a analogue and on day 9 the device was removed. Forty-eight hours after the device removal (day 11) a synthetic analogue of GnRH was administered and the animals were fixed-time artificially inseminated at the time of GnRH injection. The inseminations were performed using four different batches from the same Holstein bull. Among the heifers that were synchronized (87.2%), 30.8% ovulated until 24 h after TAI and 56.4% ovulated between 24 and 32 h after TAI. The conception rate was 61.5%. No effects of ovulation time in conception rates were detected. The conception rate from heifers that ovulated until 24 h after TAI was 58.3% and from heifers that ovulated between 24 and 32 h after TAI was 77.3%. The mean of ovulatory follicle in heifers that ovulated until 24 h was 14.3 mm and in heifers that ovulated between 24 and 32 h was 11.9 mm.Discussion: Taking together, the findings of the present study, along with those of others, emphasize the concept that development of practical methods for TAI offers significant advantages to dairy producers if conception rates are close or greater to those obtained after breeding at detected estrus. Thus, the results of the present study reinforce the possibility of making dairy cattle production more cost-effective using TAI. In conclusion, with the progesterone-based TAI protocol of the present experiment all synchronized animals ovulated up to 32 h after GnRH+TAI and no effects of ovulation time related to conception rate was detected. The exogenous control of luteal and follicular development facilitated the reproductive management and animal handling. Also, inseminating the heifers at the moment of GnRH injection in a progesterone-based TAI protocol is a practical strategy and provided satisfactory results regarding ovulation and conception rates in dairy heifers.

Assessment of field fertility and several in vitro sperm characteristics following the use of different Angus sires in a timed-AI program with suckled Nelore cows

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Primary Pulmonary Choriocarcinoma After Human Chorionic Gonadotropin Normalization Following Hydatidiform Mole A Report of Two Cases

BACKGROUND: Primary pulmonary choriocarcinoma (PPC) is rare and frequently leads to death.CASES: Two young patients presented with previous molar pregnancy and spontaneous serum human chorionic gonadotropin (hCG) normalization. Patient 1 was referred to our center after partial response to chemotherapy. Pulmonary lobectomy was performed, and hCG rapidly declined. During further chemotherapy, liver metastasis was detected by positron emission tomography. Right hepatectomy was performed, and hCG declined for 28 days, but increased again despite chemotherapy. This patient died from hepatic failure 3 years after diagnosis. Patient 2 presented with persistently high hCG, though the affected organ was not identified. Chemotherapy was unsuccessful. Patient reevaluation showed an isolated pulmonary mass. Pulmonary lobectomy was performed; 2 weeks later, hCG was normal and consolidation with 2 cycles of chemotherapy was administered. The patient has been in remission for 24 months. PPC was confirmed by histo pathology and immunohistochemistry in both cases. Gestational origin of the tumor was confirmed by molecular genetic analysis (polymorphic microsatellite markers).CONCLUSION: The possibility of choriocarcinoma cannot be overlooked in young women with an isolated pulmonary mass. Early diagnosis, prompt chemotherapy, and surgical resection in a specialized center improves the prognosis. (J Reprod Med 2010;55:311-316)

The possible role of selenium status in adverse pregnancy outcomes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Factors affecting conception rates following artificial insemination or embryo transfer in lactating holstein cows

The objective of this study was to evaluate the factors that may affect conception rates (CR) following artificial insemination (AI) or embryo transfer (ET) in lactating Holstein cows. Estrous cycling cows producing 33.1 +/- 7.2 kg of milk/d received PGF(2 alpha) injections and were assigned randomly to 1 of 2 groups (AI or ET). Cows detected in estrus (n = 387) between 48 and 96 h after the PGF2a injection received AI (n = 227) 12 h after detection of estrus or ET (n = 160) 6 to 8 d later (1 fresh embryo, grade 1 or 2, produced from nonlactating cows). Pregnancy was diagnosed at 28 and 42 d after estrus, and embryonic loss occurred when a cow was pregnant on d 28 but not pregnant on d 42. Ovulation, conception, and embryonic loss were analyzed by a logistic model to evaluate the effects of covariates [days in milk (DIM), milk yield, body temperature (BT) at d 7 and 14 post-AI, and serum concentration of progesterone (P4) at d 7 and 14 post-AI] on the probability of success. The first analysis included all cows that were detected in estrus. The CR of AI and ET were different on d 28 (AI, 32.6% vs. ET, 49.4%) and 42 (AI, 29.1% vs. ET, 38.8%) and were negatively influenced by high BT (d 7) and DIM. The second analysis included only cows with a corpus luteum on d 7. Ovulation rate was 84.8% and was only negatively affected by DIM. Conception rates of AI and ET were different on d 28 (AI, 37.9% vs. ET, 59.4%) and 42 (AI, 33.8% vs. ET, 46.6%) and were negatively influenced by high BT (d 7). The third analysis included only ovulating cows that were 7 d postestrus. Conception rates of AI and ET were different on d 28 (AI, 37.5% vs. ET, 63.2%) and 42 (AI, 31.7% vs. ET, 51.7%) and were negatively influenced by high BT (d 7). There was a positive effect of serum concentration of P4 and a negative effect of milk production on the probability of conception for the AI group but not for the ET group. The fourth analysis was embryonic loss (AI, 10.8% vs. ET, 21.5%). The transfer of fresh embryos is an important tool to increase the probability of conception of lactating Holstein cows because it can bypass the negative effects of milk production and low P4 on the early embryo. The superiority of ET vs. AI is more evident in high-producing cows. High BT measured on d 7 had a negative effect on CR and embryonic retention.

Ultrasonographic monitoring of early pregnancy development in Murrah buffalo heifers (Bubalus bubalis)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fertility-associated antigen on Nelore bull sperm and reproductive outcomes following first-service fixed-time AT of Nelore cows and heifers

The objective was to determine whether the presence of fertility-associated antigen (FAA) on sperm collected from Nelore (Bos indicus) bulls can be used to assess potential fertility of sperm for use at first-service fixed-time AI (TAI). Six Nelore bulls were selected based on FAA status (FAA-negative: N = 3; FAA-positive: N = 3) and the ability to produce neat semen with >= 70% morphologically normal sperm and 60% estimated progressive motility before cryopmservation. In Experiment 1, suckled multiparous Nelore cows (N = 835) were evaluated for body condition score (BCS) and received an intravaginal progesterone device (CIDR) and 2.0 mg of estradiol benzoate (Day 0). on Day 9 the CIDR was removed, 12.5 mg of PGF(2 alpha) and 0.5 mg of estradiol cypionate were administered, and calves were removed for 48 h. All cows received TAI on Day II (48 h after CIDR removal). Pregnancy per TAI (P/TAI) was not different between FAA-positive and FAA-negative bulls (41.5% vs. 39.3%, respectively). There was an effect of AI technician on P/TAI (36.0% vs. 43.9%; P < 0.05) and BCS tended to affect P/TAI (P = 0.09), as cows with BCS >= 2.75 were 1.4 times more likely to become pregnant compared with cows with BCS < 2.75. In Experiment 2, nulliparous Nelore heifers (N = 617) were evaluated for BCS and received a CIDR and estradiol benzoate (2.0 mg) on Day 0. on Day 7, all heifers received PGF(2 alpha) (12.5 mg). on Day 9, CIDR inserts were removed and all heifers received estradiol cypionate (0.6 mg) and 200 IU eCG. All heifers received TAI on Day 11 (48 h after CIDR removal). Pregnancy/TAI was different (P = 0.04) between FAA-positive and FAA-negative bulls (33.7% vs. 40.7%, respectively). Presence of FAA on sperm was unsuccessful in assessing the potential fertility of sperm for use in TAI. (C) 2012 Elsevier B.V. All rights reserved.

Amniocentesis and biochemical evaluation of amniotic fluid in ewes at 70, 100 and 145 days of pregnancy

The objectives of this study were to determine the technical viability of amniocentesis in sheep and to observe biochemical changes in the amniotic fluid components. Amniotic fluid samples were collected by puncture in the greatest curvature of the uterine hum at days 70, 100, and 145 of pregnancy. The surgical procedure for collection of amniotic fluid samples was safe and efficient. For three stages of pregnancy, the following results were obtained: pH values 8.36, 7.34 and 7.37; glucose concentrations, 16.06. 8.58, and 3.79 g/dl; urea values, 42.68, 33.53, and 25.49 mg/dl; creatinine, 0.85, 5.04, and 11.25 g/dl; Gama-GT enzyme, 12.58, 14.20, and 12.30 UI/l; sodium concentrations, 146.60, 129.42, and 103.8 mmol/l: potassium concentrations, 9.79, 6.15, and 8.65 mmol/l; chloride, 96.59, 85.28, and 65.35 mmol/l; total protein, 0.14, 0.23, and 0.24 g/dl, respectively. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Differential response of the luteal phase and fertility in cattle following ovulation of the first-wave follicle with human chorionic gonadotropin or an agonist of gonadotropin-releasing hormone

A series of experiments with Holstein heifers was conducted to develop the capability of inducing accessory corpus luteum (CL) with a GnRH agonist (Buserelin, 8 mu g; GnRHa) or hCG; (3,000 IU) to increase plasma progesterone concentrations (Exp. 1, 2, and 3) and to test whether induction of accessory CL with hCG will increase conception rates in heifers (Exp. 4) and lactating cows (Exp. 5). In Exp. 1, heifers were treated on d 5 after estrus with GnRHa (n = 8) or saline (n = 7); heifers in Exp. 2 received hCG (n = 5) or saline (n = 4) on d 5. Experiment 3 allowed a contemporary evaluation of heifers treated on d 5 with GnRHa (n = 6), hCG (n = 6), saline (n = 6), or GnRHa at d 5 and hCG at the time of the induced ovulation (n = 5). The GnRHa and hCG were equally effective in inducing an accessory CL (93% induction rate), but the subsequent increase in progesterone concentrations was greater in hCG-treated heifers. A greater half life of hCG may provide longer LH-like stimulation of the first-wave follicle and subsequent developing accessory CL or a greater luteotropic effect on the original CL. Induction of an accessory CL with hCG on d 5 or 6 after insemination did not increase pregnancy rates in fertile heifers (Exp. 4: hCG = 64.8% vs control = 62.9%; n = 243) or lactating dairy cows during summer heat stress (Exp. 5: hCG = 24.2% vs control = 23.5%; n = 201).

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.

Comparison of embryo yield and pregnancy rate between in vivo and in vitro methods in the same Nelore (Bos indicus) donor cows

To investigate why the preferred means to produce bovine embryos in Brazil has changed from in vivo to in vitro, we compared these two approaches in the same Nelore cows (n = 30) and assessed total embryo production and pregnancy rates. Without a specific schedule, all cows were subjected to ultrasound-guided ovum pick up (OPU)/in vitro production (IVP) and MOET, with intervals ranging from 15 to 45 d between procedures, respectively. To produce in vivo embryos, cows were superovulated and embryos were recovered nonsurgically from 1 to 3 times (1.4 +/- 0.6). whereas OPU/IVP was repeated from 1 to 5 times (3.2 +/- 1.2) in each donor cow during a 12-mo interval. Embryos obtained from both methods were transferred to crossbred heifers. on average. 25.6 +/- 15.3 immature oocytes were collected per OPU attempt. The average number of embryos produced by OPU/IVP (9.4 +/- 5.3) was higher (P < 0.05) than the MOET method (6.7 +/- 3.7). However, pregnancy rates were lower (P < 0.05) following transfer of IVP (33.5%) versus in vivo-derived embryos (41.5%) embryos. Embryonic losses between Days 30 and 60 and fetal sex ratio were similar (P > 0.05) between in vivo and in vitro-derived embryos. We concluded that in Nelore cows, with an interval of 15 d between OPU procedures, it was possible to produce more embryos and pregnancies compared to conventional MOET. (C) 2009 Elsevier B.V. All rights reserved.

Effects of temporary calf removal and eCG on pregnancy rates to timed-insemination in progesterone-treated postpartum Nellore cows

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)