Influence of the breed of bull (Bos taurus indicus vs. Bos taurus taurus) and the breed of cow (Bos taurus indicus, Bos taurus taurus and crossbred) on the resistance of bovine embryos to heat

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

Luteal Function and Follicular Growth Following Follicular Aspiration During the Peri-Luteolysis Period in Bos indicus and Crossbred Cattle

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

Synaptonemal complex analysis of Nellore and Gyr breeds of Bos taurus indicus

A synaptonemal complex (SC) study of specimens of Nellore and Gyr breeds of Bos taurus indicus was performed with the main objective to identify and determinate the frequency of abnormalities of SC and the frequency of cells with abnormalities. All animals analyzed had 29 autosomal bivalents and one sexual bivalent. The Nellore breed had 30.00% of cells with SC abnormalities while the Gyr breed had only 11.11%. Statistical analyses showed that there were not significant differences for the number of cells with abnormalities among the breeds studied. The subspecies Bos taurus indicus had 16.92% of cells showing abnormalities, being 62.82% of these abnormalities in zygotene and 37.18% in pachytene. Some aspects regarding the frequency of cells with abnormalities and the fertility of Nellore and Gyr breeds are discussed.

Complete replacement of the mitochondrial genotype in a Bos indicus calf reconstructed by nuclear transfer to a bos taurus oocyte

Due to the exclusively maternal inheritance of mitochondria, mitochondrial genotypes can be coupled to a particular nuclear genotype by continuous mating of founder females and their female offspring to males of the desired nuclear genotype. However, backcrossing is a gradual procedure that, apart from being lengthy, cannot ascertain that genetic and epigenetic changes will modify the original nuclear genotype. Animal cloning by nuclear transfer using host ooplasm carrying polymorphic mitochondrial genomes allows, among other biotechnology applications, the coupling of nuclear and mitochondrial genotypes of diverse origin within a single generation. Previous attempts to use Bos taurus oocytes as hosts to transfer nuclei from unrelated species led to the development to the blastocyst stage but none supported gestation to term. Our aim in this study was to determine whether B. taurus oocytes support development of nuclei from the closely related B. indicus cattle and to examine the fate of their mitochondrial genotypes throughout development. We show that indicus:taurus reconstructed oocytes develop to the blastocyst stage and produce live offspring after transfer to surrogate cows. We also demonstrate that, in reconstructed embryos, donor cell-derived mitochondria undergo a stringent genetic drift during early development leading, in most cases, to a reduction or complete elimination of B. indicus mtDNA. These results demonstrate that cross-subspecies animal cloning is a viable approach both for matching diverse nuclear and cytoplasmic genes to create novel breeds of cattle and for rescuing closely related endangered cattle.

Growth hormone mRNA expression in the pituitary of Bos indicus and Bos taurus x Bos indicus crossbred young bulls treated with recombinant bovine somatotropin

The effects of breed and of recombinant bovine somatotropin (rbST) treatment on growth hormone gene expression were studied in young bulls. The experiment was completely randomized in a [2 × 2]-factorial arrangement, using two levels of rbst (0 or 250 mg/animal/14 days), and two breed groups (Nelore and Simmental x Nelore crossbred). A CDNA encoding Bos indicus growth hormone was cloned and sequenced for use as a probe in Northern and dot blot analyses. Compared to the Bos taurus structural gene, the Bos indicus CDNA was found to begin 21 bases downstream from the transcription initiation site and had only two discrepancies (C to T at position 144-His and T to C at position 354-Phe), without changes in the polypeptide sequence. However, two amino acid substitutions were found for Bubalus spp., which belong to the same tribe. The rbst treatment did not change any of the characteristics evaluated (body and pituitary gland weights, growth hormone MRNA expression level). Crossbred animals had significantly higher body weight and heavier pituitaries than Nelore cattle. Pituitary weight was proportional to body weight in both breed groups. Growth hormone MRNA expression in the pituitary was similar (P>0.075) for both breed and hormonal treatment groups, but was 31.9% higher in the pure Nelore group, suggesting that growth hormone gene transcription regulation differs among these breeds.

Follicular dynamics and plasma FSH and progesterone concentrations during follicular deviation in the first post-ovulatory wave in Nelore (Bos indicus) heifers

The objective of the present study was to characterize ovarian follicular dynamics and hormone concentrations during follicular deviation in the first wave after ovulation in Nelore (Bos indicus) heifers. Ultrasonographic exams were performed and blood samples were collected every 12 h from the day of estrus until 120-144 h after ovulation in seven females. Deviation was defined as the point at which the growth rate of the dominant follicle became greater than the growth rate of the largest subordinate follicle. Deviation occurred approximately 65 h after ovulation. Growth rate of the dominant follicle increased (P < 0.05) after deviation, while growth rate of the subordinate follicle decreased (P < 0.05). Diameter of the dominant follicle did not differ from the subordinate follicle at deviation (approximately 5.4 mm). The dominant follicle (7.6 mm) was larger (P < 0.05) than the subordinate follicle (5.3 mm) 96 h after ovulation or 24 h after deviation. Plasma FSH concentrations did not change significantly during the post-ovulatory period. The first significant increase in mean plasma progesterone concentration occurred on the day of follicular deviation. In conclusion, the interval from ovulation to follicular deviation (2.7 days) was similar to that previously reported in B. taurus females, but follicles were smaller. Diameters of the dominant follicle and subordinate follicle did not differ before deviation and deviation was characterized by an increase in dominant follicle and decrease in subordinate follicle growth rate. Variations in FSH concentrations within 12-h intervals were not involved in follicular deviation in Nelore heifers. © 2006 Elsevier B.V. All rights reserved.

Morfogênese do testículo de embriões e fetos de vacas da raça nelore (Bos taurus indicus)

The aim of this study was to accompany the process of testicular development from the non-differentiable phase to its complete formation. Embryos and fetuses of Nelore breed cows (Bos Taurus indicus) were obtained in slaughterhouses near the Uberlandia city, Minas Gerais. The gonads and the embryos were fixed in Bouin's fixative and afterwards processed for conventional optical microscopy. The gonadal was observed firstly in a 1.0 cm long embryo. In 2.5 cm long embryos the presence of the albuginea allows the sex identification. The mean thickness of the albuginea ranged from 29.08 to 558.45 mm. Gradually increase of vascularization of the albuginea and parenchyma is observed. The mediastinum is located centrally. There was a decrease in the space occupied by the testicular cords, from 63.71 to 41.99% of the total testes volume. Its diameter ranged from 31.68 to 48.80 mm. The diameter of germinal cells (and their nuclei) was from 12.27 (6.65) to 16.95 914.21) mm. The quantity of germinal cells by cross section of cord decreased from a maximum of 2.80 to 0.76. The total number of germinal cells was from 16 at the beginning of colonization of the gonad to 18.32 x 106 at the end of the study. The number of Sertoli's cells by cross section of cord ranged from 10.00 to 16.25. The results obtained show that the origin and formation of testes in embryos and fetuses from Nelore breed cows (Bos taurus indicus) does occur in a very similar way to what is described for Bos taurus taurus.

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.