Ricardo Macedo Gregory

Two-dimensional polyacrylamide gel electrophoresis of equine seminal plasma proteins and their relation with semen freezability

The objective was to evaluate protein profiles of equine seminal plasma using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and to determine whether any of these proteins were related to semen freezability. Seminal plasma was collected from 10 stallions, of high and low semen freezability, housed at the State Stud of Lower Saxony, and routinely used in AI programs. Twenty-five protein spots were identified from the two-dimensional gel (12%), seven of which were present in all samples (all proteins were identified by MALDI-MS). Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has been used to generate ion images of samples in one or more mass-to-charge (m/z) values, providing the capability of mapping specific molecules to two-dimensional coordinates of the original sample. Of the 25 proteins identified, two spots had greater relative content (P < 0.05) in seminal plasma samples collected from stallions with high semen freezability: spot 5 (80-85 kDa, isoelectric point [pI] 7.54), identified as CRISP-3; and spot 45 (18.2 kDa, pI 5.0-5.2), identified as HSP-2. Conversely, protein content was greater (P < 0.05) in seminal plasma samples from stallions with low semen freezability: spot 7 (75.4 kDa, pI 6.9-7.4), identified as lactoferrin; spot 15 (26.7 kDa, pI 5.51), identified as kallikrein; spot 25 (25 kDa, pI 7.54), identified as CRISP-3; and spot 35 (13.9 kDa, pI 3.8-4.2), identified as HSP-1. In conclusion, there were differences in the seminal plasma protein profile from stallions with high and low semen freezability. Furthermore, CRISP-3 and HSP-2 were potential seminal plasma markers of high semen freezability.

Relação entre a condição corporal e a idade das ovelhas no encarneiramento com a prenhez

In this study, the relationship between score of body condition (BC) at the beginning of mating and ewes age with the pregnancy rate (PR) of a Corriedale sheep flock, grazed in the State of Rio Grande do Sul- Brazil are presented and discussed. The BC and PR observed on the flock were 90.4% and 2.84(±0.57) respectively. The mean of BC of the pregnant (P) and non-pregnant (NP) ewes was P 2.86 (±0.56) and NP 2.64(±0.59). The values of P and NP have shown statistic significance (P< 0.001). The results showed, also, that the PR increase as the BC increases, reaching 92 and 98% on the group of ewes with BC 3.0 and 4.0 respectively. Finally, the regression logistic analysis from the data showed that the age of the ewes have no relation with the PR and a positive relationship between pregnancy and body score condition ( p=0.002).

Prenhez em rebanhos ovinos do Rio Grande do Sul-Brasil

In this paper, data from real-time ultrasound pregnancy diagnosis carried out in 45 sheep flock during six reproductive seasons are presented. A total of 27089 ewes from commercial flocks, mated during autumn, were scanned. The mean pregnancy percentage(PP) found was 81.6% showing a variation from 77.3 to 89.9%. Mutton breed flocks (Hampshire Down, Suffolk and Texel) showed the highest PP of 85.6%, followed by cross breeds, with a PP of 82.9%. The PP of wool breeds flocks (Australian Merino, Corriedale and Polwarth) was 80.8%. The causes of low PP founded on some flocks is presented and discussed.

Efeito da sincronização e da indução de estros em novilhas sobre a prenhez e o índice de repetição de crias na segunda estação reprodutiva

This research was aimed at verifying the pregnancy rate in beef heifers submitted to a protocol for estrus induction and synchronization on the parturition period and its effect on the repetition of pregnancy at the second reproductive season. The experimental animals where composed of 194 Hereford and Braford heifers divided in two groups. The treatment group included a protocol of estrus observation and synchronization and fixed-time insemination (FTAI). The control group was submitted to a conventional artificial insemination management, with estrus observation and insemination in the next turn. Both groups were submitted to a natural mating period with clean-up bulls. The heifers were also observed as primiparous on the next reproductive season. The pregnancy rate was 91.7% in the first season for both groups. In the treatment group, 82% of the treated cows calved on the first 40 days of the groups calving season, against 51.7% of the control animals, resulting in a better conception rate in treated heifers (73 vs. 55%) in the second breeding season. The estrus synchronization and FTAI management on the first breeding season of beef heifers produce better pregnancy results also on the second breeding season.

Indução da atividade cíclica ovariana pós-parto em vacas de corte submetidas à interrupção temporária do aleitamento associada ou não ao tratamento com norgestomet-estradiol

The present study aims to compare the effects of removing the calf for 72 hours associated or not with Norgestomet (NOR) implant and estradiol upon estrus induction and fertility in postpartum aciclic beef cows. The temporary weaning (DT) group (n=38) was separated from their calves for 72 hours The NOR group (n= 29) received on Day 0 a 3mg Norgestomet implant subcutaneously and 5 mg of estradiol valerate, and 3mg of Norgestomet intramuscular. On Day 9 the implants were removed and the calves were separated for 72 hours. The interval from weaning to estrous was significantly shorter (P<0.05) for cows treated with progestogens (50.1 hours) compared with calf removal group (86.4 hours). The estrous induction rate was significantly higher (P<0.05) for Norgestomet and estradiol treated cows, but ovulation rate, first pregnancy rate and at the end of the breeding season were similar for both groups. The Norgestomet and estradiol treatment of aciclic beef cows improved the estrous incidence after 72 hours tempory weaning. The results demonstrate that the treatment of aciclic beef cows with Norgestomet and estradiol increases the induction of estrous after 72 hours of temporary weaning, without interfering with the pregnancy rates of first estrous and in the reproductive season.

Resposta reprodutiva de vacas de corte associada a marcadores moleculares relacionados à fertilidade

The association between the reproductive performance, expressed by pregnancy rate at fixed timed artificial insemination and birth rate in the subsequent season in beef cows, and molecular markers linked to genes for IGF-1 receptor, LHβ, leptin, and FSH and LH receptors were evaluated. Data from 249 Aberdeen Angus adult cows were used in this study. One hundred and ninety-nine cows were subjected to four different protocols for FTAI, followed by clean-up bulls and 50 cows formed the control group, matted only with bulls for 90 days during the mating season. Body condition score (BCS) and ovarian condition score (OCE) were evaluated at the beginning of the breeding season. The birth rate in the following year was 75.5%, with no treatments influence. The BCS has influenced the birth rate, respectively 55.6%, 75.8% and 82.4% (P<0.05) for animals with BCS less than 2.5; 2.5 to 2.9; and greater than or equal to 3.0, at the beginning of the breeding season. The markers related to IGF-1 receptor gene (AFZ-1 and HEL5) were associated with the birth rate in beef cows. Cows homozygous for AFZ-1 marker showed 84.4% of birth rate, while heterozygous cows showed 71.5% (P <0.05). The presence of allele *161 to the HEL5 marker was negative on birth rate. Cows with this allele had only 33.3% of birth rate, while cows without this allele had 76.5% of birth rate (P <0.05). These results demonstrate a significant association between the markers involved with the IGF-1 receptor and reproductive performance of Aberdeen Angus beef cows.

Heterozigose individual e materna sobre o ganho de peso do nascimento ao desmame de terneiros Pampiano-Braford

Data on preweanning gain (GNDa) of Pampiano-Braford animals raised in Rio Grande do sul were analyzed to verify the influence of individual (HI) and maternal (HM) heterosis in GNDa, and to quantify it. Data were adjusted for dam age (IV), julian birth date (DJN)and calf age at weaning,(IT). However, exploratory analysis have suggested that the adjustment factors for IV, DJN, and IT are not doing this adjustment correctly (P<0.01). Because of the structure and connectedness of the major data set, the analysis was done in two different ways. In the first phase, HI and HM were estimated simultaneously, in file 2, compose of 4186 animals, and the values obtained were 31.91 and 33.53kg, respectively. In the second phase, HI was estimated in a subset of file 2 (subset 1) compose of 1507 animals with maternal heterozigosis 0.0000; HM was estimated in subset 2 of file 2 with 2777 animals with individual heterozigozis 0.5000. The values found were 13.79 and 34.08kg, respectively. All the values found for heterosis on GNDa were significant (P <0.01), and the relation between heterosis and heterozigosis for this characteristic was linear (P< 0.01). It was show that the Hereford animals, in this study, had greater genetic additive values than crossbreeds, but this difference was not sufficient to supress the effects of heterosis on GNDa. In comparison with the values used in GenSys Method, the HI and HM values, obtained in this study produced a change in the ranking of calves at preweaning, but the number of animals chosen in common by the two methods did not differ much. The cows with 1/2 breed composition had the best performance on GNDa, while the calves with maximum heterosis (breed composition 1/2) were excelled by those 3/8, when HM was included in the performance, emphasizing the importance of HM on GNDa.

Leite em pó desnatado não inativado e leite desnatado uht para preservação e fertilidade do sêmen eqüino resfriado

The present study was designed aiming to verify the need of inactivating reconstituted nonfat dry milk and the use of skim milk UHT as extender for cooling equine semen. Two experiments were performed. In the first experiment, four diluents were tested: reconstituted not-inactivated dry skim milk, inactivated reconstituted dry skim milk, UHT skim milk A and UHT skim milk B. Twenty semen collections were obtained. semen were cooled at +4°C and evaluated for progressive and total motility 0 , 24 and 48 h after collection. In the second experiment, one hundred and one cycles were studied. Mares were inseminated with semen diluted in inactivated and not-inactivated dry skim milk. Dilution was done in a 1:2 (semen:diluem) ratio and the minimal inseminating dosis used was 500x106 spermatozoa. It was concluded that it is not necessary to inactivate nonfat dry milk for using it to dilute equine semen and that skim milk UHT may be used to dilute and preserve chilled equine semen.

Ultrastructural and histological characteristics of the endometrium during early embryo development in mares

The aim of this study was to evaluate ultrastructural and histological changes in the endometrium on days 7, 10 and 13 post-ovulation in pregnant and cyclic mares. Mares were routinely examined by transrectal palpation and ultrasonographic examination of the reproductive tract until estrus was detected. In the first cycle, endometrial biopsies from 30 cyclic mares (Cyclic group) were collected on days 7, 10 and 13 post-ovulation. In the second cycle, the same mares were bred by a fertile stallion. At days 7, 10 and 13 post-ovulation intrauterine biopsies were collected. Immediately after sample collection, the mares uteri were flushed, and those mares with embryo recovery were assigned to the Pregnant group. From ovulation detection until day of uterine biopsy, blood samples to measure Progesterone concentrations were collected daily in cyclic and pregnant mares. A larger blood vessel caliber was observed in pregnant mares than in cyclic from day 7-13. On the 7th day of pregnancy a large loss of ciliated cells was evident in the group of pregnant mares in comparison with the Cyclic group and the superficial cells of the endometrium were more protruded, and a small amount of histotrophic material between the folds was observed. On the 10th day of pregnancy, the glandular histotrophic secretion and the secretion of luminal epithelium became more intense than the secretion of cyclic mares. On the 13th day of pregnancy, a very large amount of histotroph was observed within large glandular openings surrounded by ciliated cells. The concentrations of P4 were affected by day (P < 0.001), but were not affected by group. Changes occurred in the uterine environment thereupon the entry of the embryo into the uterus. In the stroma and in the lumen, these modifications may aid to provide the necessary nutrition for the initial development of the embryo and to promote changes at cellular structures that will interact in the embryonic signaling and future fixation, implantation and placentation.

Early Embryo Development in Mares: Proteomics of Uterine Fluid

The expression and regulation of endometrial proteins is crucial for conceptus implantation and development. However, little is known about site-specific proteomic profiles of the mammalian endometrium during the peri-implantation period [1]. The conceptus-derived signals used for pregnancy recognition have been identified in nearly all of the important domestic animal species; one notable exception is the horse [2]. Proteomic descriptions of the uterine fluid of early pregnant mares are scant. The aim of this study was to compare the proteomic profile of uterine fluid on days 7, 10 and 13 after ovulation in pregnant and cyclic mares.