Incidence and transplacental transmission of Neospora caninum in primiparous females from Bos indicus slaughtered in Presidente Prudente, São Paulo, Brazil

To produce an epidemiological map of neosporosis in Brazil and identify the types of transmission of this disease, the present study evaluated the occurrence of Neospora caninum in Nelore cattle (Bos indicus) in Presidente Prudent, west region of Sao Paulo state; its vertical transmission; and the early stage in which fetuses are infected. To achieve this, serum samples from 518 slaughtered pregnant heifers and their fetuses were tested by ELISA technique and fetal brain tissues subjected to PCR. One hundred and three heifers (19.88%) had antibodies to N. caninum, as well as 38 (36.8%) of fetuses from 4 months of gestation. The conventional PCR failed to detect N. caninum DNA. These findings show that neosporosis occurs in the area studied and that it may be transmitted the transplacental route, althought N. caninum had not detected in brain tissue from non-aborted fetuses. The use of nested PCR it would be applied to increase the sensitivy of test.

Perfil de FSH e LH na divergência folicular em novilhas Nelore (Bos indicus)

Present study aimed to evaluate gonadotropins profiles in 12 Nelore heifers, in order to test the hypothesis that FSH concentrations decrease and LH presents a transient increase during follicle selection. Blood samples from jugular vein were harvested twice daily starting at the time of ovulation (D0) until D5. Plasma samples were assayed for FSH and LH by double antibody radioimmunoassay method. LH and FSH assay sensitivity was 0,02ng/ml and 0,005ng/ml, respectively. The intraassay coefficient of variation was 13,6% and 18,8%, respectively. Data (mean±SEM) were normalized to follicle deviation and analyzed by ANOVA and by linear, cubic, and quadratic regressions. Comparisons between higher and lower FSH values were also performed by T-test. There was no effect of time in plasmatic FSH and LH circulating levels when variance analysis or regression analysis were performed. However, by T-test, FSH concentrationsreached the lowest plasmatic levels 36 (0,40±0,05ng/ml) and 60 hours (0,42±0,04ng/ml) after follicular deviation, comparatively to 36 hours before deviation, when the concentrations were maximal (0,63±0,08ng/ml). In conclusion, there is a FSH decrease, although a transient LH elevation has not been confirmed encompassing follicle deviation in Nelore females.

Quantificação do interferon-tau durante o reconhecimento materno da gestação em fêmeas bovinas Bos taurus indicus

During the critical period of the maternal recognition, which occurs between days 15 and 19 of pregnancy, the conceptus must competently synthesize molecules capable of blocking the synthesis of prostaglandin F2α (PGF2α) and luteolysis. In cattle, the major macromolecule involved in suck blockage is the protein interferontau (IFN-τ). During the critical period, failures in the recognition of pregnancy determine embryonic mortality on up to 40% of inseminated cows. Data about IFN-τ in Bos taurus indicus are still scarce. Objective of this study was to quantitatively evaluate the presence of IFN-τ during the critical period for maternal recognition of pregnancy in uterine flushings obtained in vivo by Foley catheter (Days 14, 16 and 18 post estrus) or post-mortem (Day 18 post estrus). Multiparous, cyclic or pregnant zebu cows (Bos taurus indicus) on days 14, 16 and 18 post estrus were used for in vivo or post mortem uterine flushing collection. In both cases, a Ringer solution was used to wash the uterus of cows. Uterine flushings were concentrated by ultrafiltration and lyophilized. Proteins were separated by one-dimensional electrophoresis (SDS-PAGE) in a 15% polyacrilamide gel. Interferontau quantification in uterine flushings was performed by western blotting and densitometry. Non-specific protein bands were observed in both in vivo and post mortem uterine flushings. Interferon-tau was detected only in uterine flushings obtained from pregnant cows post-mortem (P<0.05). Optical density of protein bands was not affected by the day of the critical period, state (cyclic or pregnant) or interaction day x state. There was no effect of the conceptus weight or progesterone concentration on the day of uterine flushing collection in the optical density of the IFN-τ protein band. It was concluded that the detection and quantification of IFN-τ in the uterine environment of zebu cows, in these experimental?conditions, is only possible in uterine flushings obtained post-mortem.

LH surge in Nelore cows (Bos indicus), after induced estrus or after ovarian superestimulation

Considering that there is limited information about the preovulatory LH surge in Zebu cattle (Bos indicus), the purpose of the present work was to assess the LH surge in Nelore cows during the estrous cycle and after ovarian superestimulation of ovarian follicular development with FSH. This information is particularly important to improve superovulatory protocols associated with fixed-time artificial insemination. Nelore cows (n = 12) had their estrus synchronized with an intravaginal device containing progesterone (CIDR-B ®) associated with estradiol benzoate administration (EB, 2.5 mg, i.m., Day 0). Eight days later all animals were treated with PGF2α (Day 8) in the morning (8:00 h) and at night, when CIDR devices were removed (20:00 h). Starting 38 h after the first PGF2α injection, blood sampling and ovarian ultrasonography took place every 4 h, during 37 consecutive hours. Frequent handling may have resulted in a stress-induced suppression of LH secretion resulting in only 3 of 12 cows having ovulations at 46.7 ± 4.9 and 72.3 ± 3.8 h, respectively, after removal of CIDR-B. Thirty days later, the same animals received the described hormonal treatment associated with FSH (Folltropin ®, total dose = 200 mg) administered twice a day, during 4 consecutive days, starting on Day 5. Thirty-six hours after the first injection of PGF2α, to minimize stress, only seven blood samples were collected at 4 h interval each, and ultrasonography was performed every 12 h until ovulation. In 11 of 12 cows (92%) the LH surge and ovulation were observed 34.6 ± 1.6 and 59.5 ± 1.9 h, respectively, after removal of progesterone source. The maximum values for LH in those animals were 19.0 ± 2.6 ng/ml (mean ± S.E.M.). It is concluded that, in Nelore cows submitted to a ovarian superstimulation protocol, the LH surge occurs approximately 35 h after removal of intravaginal device containing progesterone, and approximately 12 h before the LH surge observed after an induced estrus without ovarian superstimulation. © 2008 Elsevier B.V. All rights reserved.

Physiological bases for understanding estrous cycle differences between bos taurus and bos indicus

Background: Although there is some information in the literature discussing differences of the estrous cycle of Bos taurus and Bos indicus cattle, most of the data derive from studies performed in temperate climate countries, under environmental and nutritional conditions very different than those found in tropical countries. Moreover, the physiological basis for understanding the differences between Bos taurus and Bos indicus estrous cycles are still unknown. This review explores the physiological and metabolic bases for understanding the key differences between the Bos taurus and Bos indicus estrous cycle. Moreover, it presents recent results of studies that have directly compared reproductive variables between Zebu and European cattle. Review: The knowledge of reproductive physiology, especially the differences between Bos taurus and Bos indicus, is important for the development and application of different techniques of reproductive management in cattle. In this regard, overall, Bos indicus have a greater number of small ovarian follicles and ovulatory follicles are smaller as compared to Bos taurus. Consequently, Zebu cattle also have smaller corpus luteum (CL). Nevertheless, circulating concentrations of steroid and metabolic hormones are not necessarily higher in European cattle. In fact, some studies have shown that despite ovulating smaller follicles and having smaller CL, Bos indicus cows or heifers have higher circulating concentrations of estradiol, progesterone, insulin and IGF-I compared to Bos taurus females. In addition, there are also substantial differences between Bos indicus and Bos taurus cattle in relation to follicle size at the time of selection of the dominant follicle. Conclusion: Data from very recent studies performed in Brazil have corroborated results from previous reports that have observed substantial differences in the estrous cycle variables of Bos indicus versus Bos taurus cattle. Those differences are probably related to distinct metabolism and metabolic hormone concentrations between Zebu and European cattle. This increased knowledge will allow for the establishment of more adequate reproductive management protocols in both breeds of cattle.

Influência da raça do touro (Bos indicus x Bos taurus) na tolerância ao estresse térmico calórico de embriões bovinos produzidos in vitro

To better understand the differences related to HS resistance between Bos indicus and Bos taurus, we aim to verify if the HS tolerance is due mostly to the genetic contribution from the oocyte, spermatozoa or both. Oocytes from Nelore and crossbreed Holstein cows (cHST) were collected, matured and fertilized with semen from Nelore (N), Angus (An), Brahman (Bra) and Gir (Gir) bulls. Nine six hours post insemination (hpi), ≥ 16 cells embryos were separated in two groups: control and HS. In control group, embryos were cultured at 39°C, whereas in the HS group, embryos were subjected to 41°C for 12 h, and then returned to 39°C. There was no effect of HS on blastocyst and hatched blastocyst rates in all breeds analyzed. The percentage of oocytes that cleaved and reached morula stage was significantly lower (p < 0.05) in cHST x Gir as compared to the other breeds. Additionally, blastocyst rates was higher in cHST x N than in cHST x An and cHST x Gir (p < 0.05). It was concluded that the oocyte is more important than the spermatozoa for the development of thermotolerance, since the breed of the bull did not influence embryo development after HS.

Diferenças sazonais no padrão endócrino e ovariano do ciclo estral de novilhas bos taurus indicus (Nelore)

Estrous cycle of eight Nelore heifers were evaluated during different seasons of the year (autumn n=11; winter n=8; spring n=9 and summer n=9) with daily count and measurement of follicles ≥3mm, blood was collected every 12h for LH and progesterone (P4), and after estrous every 3h for LH peak. Five ovariectomized heifers were injected with 17β-estradiol (2μg/kg) every season and blood samples collected every 3h (for 30h) thereafter for LH quantification. The monthly percent body weight difference (Δ%) did not vary among seasons. P4 concentration was higher (p<0.01) and follicle number lower during autumn and summer compared to winter and spring. During winter there were more estrous cycles with three and during summer only cycles with two follicular waves (p<0.01). As LH secretion did not vary despite P4 concentration and as there was negative correlation between higher P4 values and daily percentile variation of photoperiod (Δ%, p<0.01; r= -0.45) it is possible to suppose that there is seasonal variation on luteal cell sensitivity to LH. In the ovariectomized Nelore heifers, the LH basal concentration (without estradiol stimulus, p=0.02) and the LH response to estradiol (p<0.01) were lower during summer, leading to the hypothesis that there is seasonal variation of hypothalamic sensitivity to estradiol. According to the present experiment there are suggestions of seasonal reproduction in Nelore heifers.

Random regression models using Legendre polynomials or linear splines for test-day milk yield of dairy Gyr (Bos indicus) cattle

Studies investigating the use of random regression models for genetic evaluation of milk production in Zebu cattle are scarce. In this study, 59,744 test-day milk yield records from 7,810 first lactations of purebred dairy Gyr (Bos indicus) and crossbred (dairy Gyr × Holstein) cows were used to compare random regression models in which additive genetic and permanent environmental effects were modeled using orthogonal Legendre polynomials or linear spline functions. Residual variances were modeled considering 1, 5, or 10 classes of days in milk. Five classes fitted the changes in residual variances over the lactation adequately and were used for model comparison. The model that fitted linear spline functions with 6 knots provided the lowest sum of residual variances across lactation. On the other hand, according to the deviance information criterion (DIC) and Bayesian information criterion (BIC), a model using third-order and fourth-order Legendre polynomials for additive genetic and permanent environmental effects, respectively, provided the best fit. However, the high rank correlation (0.998) between this model and that applying third-order Legendre polynomials for additive genetic and permanent environmental effects, indicates that, in practice, the same bulls would be selected by both models. The last model, which is less parameterized, is a parsimonious option for fitting dairy Gyr breed test-day milk yield records. © 2013 American Dairy Science Association.

Effects of heat stress on development, quality and survival of Bos indicus and Bos taurus embryos produced in vitro

Heat stress is an important cause of poor development and low survival rates in bovine embryos. Experiments were conducted to test the hypothesis that Bos indicus embryos are more resistant to heat stress than are Bos taurus embryos. In experiment 1, Nelore and Jersey embryos from oocyte pick-up-derived oocytes were submitted to heat stress (96 hours post-insemination, 41 °C, 6 hours), developmental ratios were assessed at Day 7 (Day 0 = day of fertilization), and blastocysts were frozen for RNA extraction. Experiment 2 evaluated expression of COX2, CDX2, HSF1, and PLAC8 in previously frozen blastocysts. In experiment 3, Nellore and Angus embryos from oocyte pick-up-derived oocytes were submitted to heat stress (96 hours post-insemination, 41 °C, 12 hours) and transferred to recipients on Day 7. In experiment 4, embryos developed as in experiment 3 were fixed for Terminal deoxynucleotidyl transferase dUTP nick end labeling labeling and total cell counting. In experiment 1, heat stress decreased the percentage of Jersey oocytes that became blastocysts, but had no effect on Nellore embryos (34.6%, 25.0%, 39.5%, and 33.0% for Jersey control, Jersey heat-stressed, Nellore control, and Nellore heat-stressed oocytes, respectively; P < 0.05). In experiment 2, heat stress decreased (P < 0.05) expression of CDX2 and PLAC8, with higher expression of these genes in Nellore embryos than in Jersey embryos. Heat stress also decreased (P < 0.05) expression of COX2 in Jersey embryos, but had no effect on Nellore embryos. Expression of HSF1 was decreased (P < 0.05) by heat stress in both breeds, with a greater effect in Nellore embryos. In experiment 3, heat stress tended (P = 0.1) to decrease the percentage of pregnancies among cows (Day 30 to 35) that received Angus embryos. In experiment 4, heat stress increased (P < 0.05) the percentage of apoptotic blastomeres, but had no breed-specific effects. In addition, Nellore embryos had fewer (P < 0.05) Terminal deoxynucleotidyl transferase dUTP nick end labeling- positive blastomeres than did Angus embryos. We concluded that the detrimental effects of heat stress were dependent upon embryo breed and were more evident in Bos taurus embryos than in Bos indicus embryos. © 2013 Elsevier Inc.

Comparison of random regression models to estimate genetic parameters for milk production in Guzerat (Bos indicus) cows

Random regression models have been widely used to estimate genetic parameters that influence milk production in Bos taurus breeds, and more recently in B. indicus breeds. With the aim of finding appropriate random regression model to analyze milk yield, different parametric functions were compared, applied to 20,524 test-day milk yield records of 2816 first-lactation Guzerat (B. indicus) cows in Brazilian herds. The records were analyzed by random regression models whose random effects were additive genetic, permanent environmental and residual, and whose fixed effects were contemporary group, the covariable cow age at calving (linear and quadratic effects), and the herd lactation curve. The additive genetic and permanent environmental effects were modeled by the Wilmink function, a modified Wilmink function (with the second term divided by 100), a function that combined third-order Legendre polynomials with the last term of the Wilmink function, and the Ali and Schaeffer function. The residual variances were modeled by means of 1, 4, 6, or 10 heterogeneous classes, with the exception of the last term of the Wilmink function, for which there were 1, from 0.20 to 0.33. Genetic correlations between adjacent records were high values (0.83-0.99), but they declined when the interval between the test-day records increased, and were negative between the first and last records. The model employing the Ali and Schaeffer function with six residual variance classes was the most suitable for fitting the data. © FUNPEC-RP.

Physiology and endocrinology symposium: Influence of cattle genotype (Bos indicus vs. Bos taurus) on oocyte and preimplantation embryo resistance to increased temperature

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

Differential expression of members of the IGF system in OPU-derived oocytes from Nelore (Bos indicus) and Holstein (Bos taurus) cows

The insulin-like growth factor (IGF) system is related to quality of oocytes and embryos. The aim of this study was to investigate the mRNA levels of IGF1 and IGF2 and their receptors, IGFR1 and IGFR2, as well as IGFBP2, IGFBP4, and PAPP-A in oocytes from Nelore compared to Holstein cows. Pools of oocytes (20 oocytes/pool) from Nelore (n=8 pools) and Holstein (n=4 pools) were obtained via ovum pick-up (OPU, 10 sessions) and cumulus cells and zona pellucida were removed. The pools were submitted to total RNA extraction. Expression of members of the IGF system was assessed by real time RT-PCR. The mRNA expression of IGF1 and IGF2, IGFR1 and IGFR2, IGFBP2 and IGFBP4 was significantly higher (P<0.01) in oocytes from Holstein whereas the expression of PAPP-A was significantly higher (P<0.05) in oocytes from Nelore cows. The high PAPP-A expression and the low expression of IGFBP2 and IGFBP4 are associated with more efficient degradation of IGFBPs, which results in greater bioavailability of IGF in Nelore oocytes when compared to the Holstein. © 2013 Elsevier B.V.

Expression of genes related to quality of Longissimus dorsi muscle meat in Nellore (Bos indicus) and Canchim (5/8 Bos taurus×3/8 Bos indicus) cattle

This study was performed to compare CAPN1, CAPN2, CAST, TG, DGAT1 and LEP gene expressions and correlate them with meat quality traits in two genetic groups (Nellore and Canchim) in order to assess their expression profile and use their expression profile as genetic markers. We analyzed 30 young bulls (1. year old), 15 of each genetic group. Samples of the Longissimus dorsi muscle were collected for analysis of: total lipids (TL) and meat tenderness measured as Warner-Bratzler shear force (SF) and myofibrillar fragmentation (MFI) at day of slaughter and 7. days of aging. Gene expression profiles were obtained via RT-qPCR. TL and MFI showed differences between breeds, higher MFI in Canchim and higher TL in Nellore. Calpains showed no differential expression between groups, as did DGAT1, TG, and LEP. CAST was expressed more in the Nellore cattle. The only significant within-breed correlation (0.79) between gene expression and meat traits was found for DGAT1 and MFI in Canchim breed. Although the number of animals used in this study was small, the results indicate that the increased expression of CAST in Nellore may reflect tougher meat, but the lack of correlations with the meat traits indicates it is not a promising genetic marker. © 2013 Elsevier Ltd.

Localization patterns of steroid and luteinizing hormone receptors in the corpus luteum of Nelore (Bos taurus indicus) cows throughout the estrous cycle

The aim of the present study was to detect progesterone receptors (A and B isoforms), α and β estrogen receptors, luteinizing hormone receptors and aromatase cytochrome P450 enzymes in the corpus luteum of Nelore (Bos taurus indicus) cows using immunohistochemistry. The estrous cycles of 16 Nelore cows were synchronized, and luteal samples were collected via an incision into the vaginal vault. Samples were collected during specific days of the estrous cycle (days 6, 10, 15 and 18) and 24. h after circulating progesterone dropped, after luteolysis had occurred. After each biopsy was taken, all animals were resynchronized so that each biopsy was performed during a different estrous cycle. Our results showed that the concentration of studied proteins vary throughout the bovine estrous cycle. The highest concentration of α and β estrogen receptors and the highest concentration of plasma progesterone were both observed on days 10 and 15 of the estrous cycle. The highest concentration of progesterone receptors was observed on days 6 and 10 of the estrous cycle, and the most intense immunostaining for cytochrome P450 aromatase enzymes was observed on day 10 of the estrous cycle. The highest score of cells with plasma membrane immunostaining for LH receptors was observed on day 15 of the estrous cycle. In conclusion, this study demonstrates the varying concentrations of specific proteins within the corpus luteum of Nelore cows during the estrous cycle. This finding suggests that these receptors and enzymes, and their interactions, are important in regulating luteal viability. © 2013 Elsevier B.V.

Desenvolvimento do omaso de bovinos da raça Nelore (Bos taurus indicus) durante o período pré-natal

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