Relationships between growth of the preovulatory follicle and gestation success in lactating dairy cows

This report summarizes three studies conducted with lactating dairy cows aiming to increase pregnancy rates to fixed time artificial insemination (TAI) protocols. Experiment 1 was designed to determine if changing the timing of PGF2 alpha treatment during an E2/P4-based program would affect fertility to TAI or fixed-time embryo transfer (TET). In experiment 2, pregnancy rates to AI were compared following synchronized ovulation using two protocols that have been developed to reduce the period between follicular wave emergence and TAI. The Ovsynch-type protocol utilizes GnRH to synchronize the follicular wave by inducing ovulation of a dominant follicle at the beginning of the protocol, and to synchronize ovulation at the end of the protocol allowing TAI. In contrast, E2/P4-based protocols utilize E2 products in the presence of P4 to induce atresia of antral follicles and synchronize emergence of a new follicular wave. At the end of E2/P4-based protocol another E2 treatment in the absence of P4 is used to induce LH release and synchronize ovulation and allow TAI. Experiment 3 was designed to determine whether increasing the length time interval with reduced circulating P4 (proestrus) would increase fertility in a TAI program that utilized E2 and P4 to synchronize ovulation of cycling, lactating dairy cows. The overall conclusions are that circulating concentrations of progesterone and estradiol prior to and circulating concentrations of progesterone following ovulation can affect fertility in cattle. In addition, small increases in P4 concentrations near the time of AI, due to lack of complete CL regression, result in reductions in fertility. Earlier treatment with PGF2 alpha should allow greater time for CL regression, an increase in estradiol and subsequent reductions in circulating P4 that could be critical for fertility. Optimization of follicle size in TAI programs is clearly an intricate balance between oocyte quality, adequate circulating E2 near AI, and adequate circulating P4 after AI.

Improving postcryopreservation survival capacity: an embryo-focused approach

The major challenge for a greater dissemination of in vitro produced (IVP) bovine embryos is to improve embryonic survival after cryopreservation. The involvement of embryonic lipids on this issue is well documented. However, it has been recognized that not only the amount of lipids that affects embryo cryotolerance, but the embryo survival capacity after cryopreservation is a rather multifactorial event. In this review, some strategies to improve embryonic lipid composition and postcryopreservation survival by modifying the embryos themselves to make them more cryopreservable are overviewed. The use of semi-defined and defined serum-free culture media, the addition of some chemicals in the culture media to modify embryo lipid composition, and the modulation of embryo cell membrane fluidity by cholesterol or unsaturated fatty acids added to the culture media and oocyte/embryo donor nutritional management with a diet enriched in polyunsaturated fatty acids, were described as alternatives for the improvement of IVP embryo survival after cryopreservation.

A comparison of microbiological and molecular detection of vaginal Lactobacillus spp. between mares and women

A possible explanation for endometritis in mares is ascendant contamination from the vagina. The presence of Lactobacillus spp. is considered to be important in women for a healthy vaginal environment; however, there are few studies in mares related to the presence of Lactobacillus in the vaginal flora of healthy mares. The present work aims to determine the occurrence of Lactobacillus spp. in the vaginal micro-environment of mares. A total of 35 crossbred multiparous mares, aged between 4 and 12 years, with no history of reproductive problems and with healthy reproductive tracts, were used. Two vaginal swabs were obtained from the mares during estrus for Lactobacillus isolation and PCR evaluation. Ten human female volunteers, aged between 24 and 35 years, sexually active, with no history of gynecological diseases and treatments in the past two years were used. Lactobacillus spp. were isolated from 5.7% of the mares' vaginal samples and from 90% of the women's vaginal samples. Lactobacillus DNA was detected by PCR in 22.9% of the mares' vaginal samples and in all of the vaginal samples from the healthy women. The primers used here were demonstrated to have in silico specificity for the detection of L. equi (AB425924.1), L. pantheris (DQ471798.1) and L. mucosae (DQ471799.1), but they did not anneal on Enterococcus faecalis (EU887827.1) or E. faecium (EU887814.1). In conclusion, this study showed a low occurrence of Lactobacillus spp. in mares, suggesting that this bacterium may not play a fundamental role in the equilibrium of the vaginal micro- environment of normal mares.

The influence of insemination dose on pregnancy per fixed-time artificial insemination in beef cows is affected by semen extender

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

Da classificação biológica à classificação digital: perspectivas de renovação em classificação arquivística

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Processo judicial previdenciário: a necessária mudança de paradigma para a efetividade da política de previdencia social no Brasil

Pós-graduação em Direito - FCHS

Safety performance models for urban intersections in Brazil

This paper presents a modeling effort for developing safety performance models (SPM) for urban intersections for three major Brazilian cities. The proposed methodology for calibrating SPM has been divided into the following steps: defining the safety study objective, choosing predictive variables and sample size, data acquisition, defining model expression and model parameters and model evaluation. Among the predictive variables explored in the calibration phase were exposure variables (AADT), number of lanes, number of approaches and central median status. SPMs were obtained for three cities: Fortaleza, Belo Horizonte and Brasilia. The SPM developed for signalized intersections in Fortaleza and Belo Horizonte had the same structure and the most significant independent variables, which were AADT entering the intersection and number of lanes, and in addition, the coefficient of the best models were in the same range of values. For Brasilia, because of the sample size, the signalized and unsignalized intersections were grouped, and the AADT was split in minor and major approaches, which were the most significant variables. This paper also evaluated SPM transferability to other jurisdiction. The SPM for signalized intersections from Fortaleza and Belo Horizonte have been recalibrated (in terms of the COx) to the city of Porto Alegre. The models were adjusted following the Highway Safety Manual (HSM) calibration procedure and yielded C-x of 0.65 and 2.06 for Fortaleza and Belo Horizonte SPM respectively. This paper showed the experience and future challenges toward the initiatives on development of SPMs in Brazil, that can serve as a guide for other countries that are in the same stage in this subject. (C) 2014 Elsevier Ltd. All rights reserved.

Effect of hydrogen peroxide on thawed ovine sperm motility

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

Control of oocyte maturation

Oocyte maturation is a complex process involving nuclear and cytoplasmic maturation. The nuclear maturation is a chromosomal segregation and the cytoplasmic maturation involves the reorganization of the cytoplasmic organelles, mRNA transcription and storage of proteins to be used during fertilization and early embryo development. The mechanism of oocyte maturation in vivo and in vitro still are not totally understood. However it is generally accepted that the second messenger cyclic adenosine monophosphate (cAMP) plays a critical role in the maintenance of meiotic blockage of mammalian oocytes. A relative increase in the level of cAMP within the oocyte is essential for maintaining meiosis block, while a decrease in cAMP oocyte concentration allows the resumption of meiosis. The oocyte cAMP concentration is regulated by a balance of two types of enzymes: adenylate cyclase (AC) and phosphodiesterases (PDEs), which are responsible for the synthesis and degradation of cAMP, respectively. After being synthesized by AC in cumulus cells, cAMP are transferred to the oocyte through gap junctions. Thus, specific subtypes PDEs are able to inhibit or attenuate the spontaneous meiotic maturation of oocytes with PDE4 primarily involved in the metabolism of cAMP in granulosa cells and PDE3 in the oocyte. Although the immature oocytes can resume meiosis in vitro, after being removed from antral follicles, cytoplasmic maturation seems to occur asynchronously with nuclear maturation. Therefore, knowledge of the oocyte maturation process is fundamental for the development of methodologies to increase the success of in vitro embryo production and to develop treatments for various forms of infertility. This review will present current knowledge about the maintenance of the oocyte in prophase arrest, and the resumption of meiosis during oocyte maturation, focusing mainly on the changes that take place in the oocyte.

The physiology and impact on fertility of the period of proestrus in lactating dairy cows

In cattle, proestrus begins with the initiation of luteolysis and ends with initiation of estrus and the GnRH/LH surge. This period is marked by a dramatic decrease in circulating progesterone (P4) that reaches a nadir by about 36-48 h in cows undergoing natural or prostaglandin F2 alpha (PGF)-induced luteolysis. Inadequate luteolysis is a cause of reduced fertility particularly in timed AI programs with small elevations in circulating P4 reducing fertility. Increasing circulating estradiol (E2) during proestrus is dependent on presence, size, and function of the dominant follicle and this varies during natural proestrus, due to whether animals have two or three follicular waves, and during PGF-induced proestrus, according to stage of the follicular wave at time of PGF treatment. Inadequate circulating E2 can limit fertility and increase pregnancy loss in some specific circumstances such as in cows with low BCS and in cows during heat stress. Thus, studies to optimize the length of proestrus and the concentrations of E2 and P4 during proestrus could produce substantial improvements in fertility and reductions in pregnancy loss.

The role of proestrus on fertility and postovulatory uterine function in the cow

Modern protocols to synchronize ovulations for timed artificial insemination and timed embryo transfer that include manipulations in the proestrus period (i.e., between luteolysis and estrus) affect fertility in cattle. Specifically, stimulating pre-ovulatory follicle growth and exposure to estrogens after CL regression increase the proportion of cows pregnant and decrease late embryo mortality. Such effects may be due to both preovulatory actions of estrogens and post-ovulatory actions of progesterone, as concentrations of the later hormone may be changed in response to manipulations conducted during proestrus. In the first portion of this paper we describe strategies used recently to manipulate the proestrus period in protocols for synchronization of ovulation, and to present evidence of their effects on fertility. Manipulations of timing and prominence of sex steroids during the proestrus and early diestrus that affect fertility may act on targets such as the endometrium. This tissue expresses receptors for both estrogens and progesterone and these hormones change endometrial function to support conceptus growth and pregnancy maintenance. However, specific cellular and molecular mechanisms through which fertility is affected via manipulations of the proestrus are poorly understood. In the second portion of this paper we describe a well-defined animal model to study changes in endometrial function induced by manipulations conducted during the proestrus. Such manipulations induced endometrial changes on sex steroid receptors expression, cell proliferation, oxidative metabolism and eicosanoid synthesis in the uterus, but not on glucose transport to uterine lumen. In summary, evidence is accumulating to support a positive role of increasing duration and estrogen availability during the proestrus on fertility to synchronization protocols. Such positive effects may be through changes in endometrial function to stimulate conceptus growth and survival.

Lethal effect of high concentrations of Parecoxib and Flunixin meglumine on the in vitro culture of bovine embryos

Since cyclooxygenase (COX) inhibitors have been pointed out as potential treatments to increase pregnancy rates after embryo transfer, the present experiment aimed to evaluate the effects of flunixin meglumine (FM) and parecoxib (P), a COX-1 and 2 or COX-2 specific inhibitor, respectively, on the development of bovine embryos until the hatched blastocyst stage. In vitro produced bovine embryos were cultured in media with different concentrations of FM (0.14; 1.4; 14; 140 or 1400 mu g/ml) or P (0.09; 0.9; 9; 90 or 900 mu g/ml) and the production rates were evaluated. Concentrations of FM <= 14 mu g/ml and P <= 90 mu g/ml did not impair embryo development, although compiled data from non-lethal FM concentrations (<= 14 mu g/ml) indicated a toxic effect enough to decrease the hatching rate of blastocysts. Concentrations of FM at 140 and 1400 mu g/ml and P at 900 mu g/ml were lethal as no cleavage was detected on presumptive zygotes.

Developmental aspects of early pregnancy in mares

In mares, the embryo migrates to the uterus between days 5 and 6 postovulation beginning its mobility through all uterine segments, which is essential for the maternal recognition of pregnancy. During the mobility phase, the embryonic vesicle shows a linear growth rate until its fixation between days 15 and 17, when the orientation phenomenon occurs. From fixation to day 28 of pregnancy, the embryonic growth is less evident (plateau) by cross-section ultrasound examination. After this period the linear growth rate is reestablished until day 46. This plateau is attributed to the increased uterine tone that compresses the vesicle and to volume expansion, making it difficult to detect the conceptus growth only by the cross-section diameter. Around day 20, the embryo proper is visualized as an echogenic spot in the ventral aspect of the vesicle. Additionally, development of allantoic sac, embryonic heartbeat, yolk sac regression and posterior umbilical cord formation also can be visualized from days 20 to 40. An intimate interaction between uterus and conceptus is essential for the normal pregnancy development. Color-and spectral-Doppler ultrasonography can be useful for the evaluation of this interface. A gradual increase on uterine vascularity during the early pregnancy and transient changes in endometrial vascularity accompanying the vesicle location during the mobility phase have been described. Around day 38 of gestation, the formation of the endometrial cups begins and, consequently, the synthesis of the equine chorionic gonadotropin (eCG) induces the formation and development of supplementary corpora lutea, which are important to secrete progesterone and to maintain pregnancy until around day 120.