Análise morfológica dos folículos primordiais de ovários de ovino criopreservados em EG, DMSO e sua associaçã o

Objective: Compare the cryoprotectants Dimethyl Sulphoxide (DMSO), Ethylene Glycol (EG) and their association for cryopreservation of sheep ovarian cortex. Methodology: Fragments collected from ovaries were divided into 3 parts. 1. One part from sample was destined for analysis of fresh material. 2. The second part was incubated with solution of freezing having 1,5M EG or 1,5M DMSO or 1,5MEG + 1,5M DMSO and washed for dilution of the cryoprotectants. 3. The third part was submitted to cryopreservation using the same cryoprotectans (EG 1,5M; DMSO 1,5M and EG + DMSO 1,5M) and cryopreserved. In all groups, one part of sample was submitted to pre-antral follicles isolation and the remainder was destined to ultra-structural analysis. Results: After isolation of fresh primordial follicles (control), the percentage of viable follicles was 78,9%. The percentage of viable follicles only exposed to cryoprotectants 1,5M EG, 1,5M DMSO and 1,5M EG + 1,5M DMSO was 77,1%, 68,4% and 60,7% respectively. After cryopreservation were 75%, 60% and 55,6% respectively. Ultra-structural analysis of the primordial follicles derived from fresh ovarian fragments or from fragments just exposed to the cryoprotectants showed similar morphology. However, in frozen samples, alterations of mitochondria were observed in all groups. Despite this, the integrity of the remained organelles was preserved in follicles cryopreserved with EG, while that in others groups (DMSO and association) an excess of vacuolizaton in cytoplasm of oocytes and swelling of nuclear membrane was observed indicating degeneration. Conclusion: The Ehilene Glycol seems to be the cryoprotector more adequated for cryopreservation of sheep ovarian tissue.

Detecting Loci under Recent Positive Selection in Dairy and Beef Cattle by Combining Different Genome-Wide Scan Methods

As the methodologies available for the detection of positive selection from genomic data vary in terms of assumptions and execution, weak correlations are expected among them. However, if there is any given signal that is consistently supported across different methodologies, it is strong evidence that the locus has been under past selection. In this paper, a straightforward frequentist approach based on the Stouffer Method to combine P-values across different tests for evidence of recent positive selection in common variations, as well as strategies for extracting biological information from the detected signals, were described and applied to high density single nucleotide polymorphism (SNP) data generated from dairy and beef cattle (taurine and indicine). The ancestral Bovinae allele state of over 440,000 SNP is also reported. Using this combination of methods, highly significant (P<3.17×10-7) population-specific sweeps pointing out to candidate genes and pathways that may be involved in beef and dairy production were identified. The most significant signal was found in the Cornichon homolog 3 gene (CNIH3) in Brown Swiss (P = 3.82×10-12), and may be involved in the regulation of pre-ovulatory luteinizing hormone surge. Other putative pathways under selection are the glucolysis/gluconeogenesis, transcription machinery and chemokine/cytokine activity in Angus; calpain-calpastatin system and ribosome biogenesis in Brown Swiss; and gangliosides deposition in milk fat globules in Gyr. The composite method, combined with the strategies applied to retrieve functional information, may be a useful tool for surveying genome-wide selective sweeps and providing insights in to the source of selection.

Genome-wide scans to uncover loci underlying bovine reproductive biology

Pós-graduação em Medicina Veterinária - FCAV

Perfil gênico no oviduto bovino de fêmeas Nelore e Aberdeen angus

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

Bupropion treatment increases epididymal contractility and impairs sperm quality with no effects on the epididymal sperm transit time of male rats

Bupropion is a dopamine (DA) and norepinephrine (NE) reuptake inhibitor used as smoking cessation and antidepressant drug with a lower incidence of male sexual dysfunction. We showed previously that sibutramine, a norepinephrine/serotonine reuptake inhibitor, reduced male rat fertility. As there are no studies evaluating the impact of bupropion treatment on spermatic parameters and male fertility, we evaluated the effects of bupropion treatment (15 and 30 mg kg(-1), 30 days) on sexual behavior, spermatic parameters and fertility of male Wistar rats and on the epididymal duct in vitro contractility. Bupropion 15 mg kg(-1) increased the serum luteinizing hormone level and the epididymal duct contractility, but the sperm quality was not affected. At 30 mg kg(-1) bupropion impaired sperm quality increasing the incidence of non-progressive sperm. The male sexual behavior and fertility were not modified at both bupropion doses. These results, in rats, suggest the importance of studies evaluating the effects of bupropion on the human male sperm quality.

Oogenesis in Laetacara araguaiae (Ottoni and Costa, 2009) (Labriformes: Cichlidae)

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

Criptorquidia e exposição in utero ao di(n-butil)-ftalato e à acrilamida: avaliação morfológica do dano testicular

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Desempenho reprodutivo e diferenciação gonadal em piau-três-pintas, Leporinus friderici, Bloch 1794

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ovarian activity and plasma concentrations of progesterone and estradiol during pregnancy in jennies

The objectives of this study were to determine ovarian activity (with ultrasound) and plasma concentrations of progesterone and estradiol during pregnancy in jennies. There was considerable ovarian activity during the second month of pregnancy. Secondary corpora lutea (total of 2 to 7 per jenny) were formed (mainly by luteinization) starting on Day 41.8 +/- 1.0 (range Days 38 to 46; ovulation = Day 0). The echogenicity of the primary and secondary corpora lutea gradually decreased during pregnancy. Plasma progesterone concentrations increased between Days 0 and 10 (0.9 and 19.9 ng/mL, respectively), gradually decreased to Day 30 (12.1 ng/mL), increased between Days 30 and 40 (plateau, at approximately 17 ng/mL), gradually declined from Days 110 to 160 (nadir of approximately 6 ng/mL), and remained nearly constant until increasing again just before parturition. Plasma estradiol concentrations increased gradually from Day 65, peaked (1.2 ng/mL) on Day 165 (greater than or equal to 1 ng/mL on Days 150 to 210), and decreased thereafter, with very low concentrations during the last 20 d before parturition. Ovarian function and hormone profiles were generally similar to those previously reported during pregnancy in mares. (C) 1998 by Elsevier B.V.

Effect of timing of estradiol benzoate administration upon synchronization of ovulation in suckling Nelore cows (Bos indicus) treated with a progesterone-releasing intravaginal device

The present study investigated how the timing of the administration of estradiol benzoate (EB) impacted the synchronization of ovulation in fixed-time artificial insemination protocols of cattle. To accomplish this, two experiments were conducted, with EB injection occurring at different times: at withdrawal of the progesterone-releasing (N) intravaginal device or 24 h later. The effectiveness of these times was compared by examining ovarian follicular dynamics (Experiment 1, n = 30) and conception rates (Experiment 2, n = 504). In Experiment 1, follicular dynamics was performed in 30 Nelore cows (Bos indicus) allocated into two groups. on a random day of the estrous cycle (Day 0), both groups received 2 mg of EB i.m. and a P4-releasing intravaginal device, which was removed on Day 8, when 400 IU of eCG and 150 mu g of PGF were administered. The control group (G-EB9; n = 15) received 1 mg of EB on Day 9, while Group EB8 (G-EB8; n = 15) received the same dose a day earlier. Ovarian ultrasonographic evaluations were performed every 8 h after device removal until ovulation. The timing of EB administration (Day 8 compared with Day 9) did affect the interval between P4 device removal to ovulation (59.4 +/- 2.0 h compared with 69.3 +/- 1.7 h) and maximum diameter of dominant (1.54 +/- 0.06 a cm compared with 1.71 +/- 0.05 b cm, P = 0.03) and ovulatory (1.46 +/- 0.05 a cm compared with 1.58 +/- 0.04 b cm, P < 0.01) follicles. In Experiment 2,504 suckling cows received the same treatment described in Experiment 1, but insemination was performed as follows: Group EB8-AI48h (G-EB8-AI48h; n = 119) and Group EB8-AI54h (G-EB8-AI54h; n = 134) received 1 mg of EB on Day 8 and FrAI was performed, respectively, 48 or 54 h after P4 device removal. Group EB9-AI48h (G-EB9-AI48h; n = 126) and Group EB9-AI54h (G-EB9-AI54h n = 125) received the same treatments and underwent the same FTAI protocols as G-EB8-AI48h and G-EB8-AI54h, respectively; however, EB was administered on Day 9. Conception rates were greater (P < 0.05) in G-EB9-AI54h 163.2% (79/125) a], G-EB9-AI48h [58.7% (74/126) a] and G-EB8-AI48h [58.8% (70/119) a] than in G-EB8-AI54h [34.3% (46/134) b]. We concluded that when EB administration occurred at device withdrawal (D8), the interval to ovulation shortened and dominant and ovulatory follicle diameters decreased. Furthermore, when EB treatment was performed 24 h after device removal, FTAI conducted at either 48 or 54 h resulted in similar conception rates. However, EB treatment on the same day as device withdrawal resulted in a lesser conception rate when FTAI was conducted 54 h after device removal. (C) 2007 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.

Comparison of progesterone-based protocols with gonadotropin-releasing hormone or estradiol benzoate for timed artificial insemination or embryo transfer in lactating dairy cows

The objective was to compare two protocols for synchronizing ovulation in lactating Holstein cows submitted to timed AI (TAI) or timed ET (TET). Within each farm (n = 8), cows (n = 883; mean +/- SEM 166.24 +/- 3.27 d postpartum, yielding 36.8 +/- 0.34 kg of milk/d) were randomly assigned to receive either: 1) an intravaginal progesterone insert (CIDR (R)) with 1.9 g of progesterone + GnRH on Day -10, CIDR (R) withdrawal + PGF2 alpha on Day -3, and 1 mg estradiol cypionate on Day -2 (treatment GP-P-E; n(TAI) = 180; n(TET) = 260); or 2) a CIDR (R) insert + 2 mg estradiol benzoate on Day -10, PGF2 alpha on Day -3, CIDR (R) withdrawal + 1 mg estradiol cypionate on Day -2 (treatment EP-P-E; n(TAI) = 174; n(TET) = 269). Cows were subsequently randomly assigned to receive either TAT on Day 0 or TET on Day 7. Serum progesterone concentration on Day -3 was greater in GP-P-E than in EP-P-E (2.89 +/- 0.15 vs 2.29 +/- 0.15 ng/mL; P < 0.01), with no significant effect of group on serum progesterone on Day 7. Compared to cows submitted to TAI, those submitted to TET had greater pregnancy rates on Day 28 (44.0% [233/5291 vs 29.7% [105/354]; p < 0.001) and on Day 60 (37.6% [199/529] vs 26.5 [94/354]; P < 0.001). However, there were no effects of treatments (GP-P-E vs EP-P-E; P > 0.10) on synchronization (87.0% [383/440] vs 85.3% [378/443]), conception (TAI: 35.3% [55/156] vs 33.8% [50/148]; TET: 50.7% [115/227] vs 51.3% [118/230]) and pregnancy rates on Days 28 (TAT: 30.5% [55/180] vs 28.7% 150/174]; TET: 44.2% [115/260] vs 43.9% [118/2691) and 60 (TAI: 27.2% [49/80] vs 25.9% [45/174]; TET: 38.8% [101/260] vs 36.4% [98/269]). In conclusion, GP-P-E increased serum progesterone concentrations on Day -3, but rates of synchronization, conception, and pregnancy were not significantly different between cows submitted to GP-P-E and EP-P-E protocols, regardless of whether they were inseminated or received an embryo. (c) 2011 Elsevier B.V. All rights reserved.

Relative expression of insulin like growth factor I (IGFI) and follicle stimulating hormone receptor (FSHR) in follicles and ovarian tissue from Bos primigenius indicus (Nelore)

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

Synchronization of ovulation in crossbred dairy heifers using gonadotrophin-releasing hormone agonist, prostaglandin F2a and human chorionic gonadotrophin or estradiol benzoate

Girolando (Gir x Holstein) is a very common dairy breed in Brazil because it combines the rusticity of Gir (Bos indicus) with the high milk yield of Holstein (Bos taurus). The ovarian follicular dynamics and hormonal treatments for synchronization of ovulation and timed artificial insemination were studied in Girolando heifers. The injection of a gonadotrophin-releasing hormone (GnRH) agonist was followed 6 or 7 days (d) later by prostaglandin F2a (PGF2a). Twenty-four hours after PGF2a injection either human chorionic gonadotropin (hCG, GPh-d6 and GPh-d7 groups) or estradiol benzoate (EB, GPE-d6 and GPE-d7 groups) was administered to synchronize ovulation and consequently allow timed artificial insemination (AI) 24 and 30 h after hCG and EB injection, respectively. Follicular dynamics in Girolando heifers was characterized by the predominance of three follicular waves (71.4%) with sizes of dominant follicles (10-13 mm) and corpus luteum (approximately 20 mm) similar to those for Bos indicus cattle. In the GnRH-PGF-hCG protocol, hCG administration induced earlier ovulation (67.4 h, P<0.01) compared to the control group (GnRH-PGF) and a better synchronization of ovulation, since most of it occurred within a period of 12 to 17 h. Pregnancy rate after timed AI was 42.8 (3/7, GPh-d6) to 50% (7/14, GPh-d7). In contrast, estradiol benzoate (GnRH-PGF-EB protocol) synchronized ovulation of only 5 of 11 heifers from the GPE-d7 group and of none (0/7) from the GPE-d6 group, which led to low pregnancy rates after timed AI (27.3 and 0%, respectively). However, since a small number of Girolando heifers was used to determine pregnancy rates in the present study, pregnancy rates should be confirmed with a larger number of animals.