The interval between the emergence of pharmacologically synchronized ovarian follicular waves and ovum pickup does not significantly affect in vitro embryo production in Bos indicus, Bos taurus, and Bubalus bubalis

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

Different true-protein sources do not modify the metabolism of crossbred Bos taurus x Bos indicus growing heifers

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

Synaptomenal complex analysis of four breeds of Bos taurus taurus x B. taurus indicus hybrids

The synaptonemal complex (SC) was analyzed in four F1 hybrids of Bos taurus taurus and B. taurus indicus including Gyr-Simmental (G-S), Nelore Simmental (N-S), Gyr-Holstein-Friesian (G-H) and Nelore-Piemontese (N-P). We analysed the frequency of various types of SC abnormalities and the frequency of cells with SC abnormalities. The results were compared with similar observations made on purebred animals. All the animals studied possessed 29 autosomal and one sex bivalent. The frequency of cells with abnormalities in the hybrids were 28.0% in the N-P, 29.1% in the G-S, 33.3% in the N-S and 40.0% in the G-H. The frequency of cells with abnormalities in the four hybrids was 31.5%; 57.9% of these abnormalities occurred in zygotene and 42.0% occurred in pachytene. The comparisons among the hybrids and among the hybrids and their parental breeds showed that the only significant difference was between Gyr and Gyr-Holstein-Friesian animals. Some aspects of the relationship between the frequency of cells with anomalies and the fertility of hybrids are discussed.

Imputation of microsatellite alleles from dense SNP genotypes for parentage verification across multiple Bos taurus and Bos indicus breeds

To assist cattle producers transition from microsatellite (MS) to single nucleotide polymorphism (SNP) genotyping for parental verification we previously devised an effective and inexpensive method to impute MS alleles from SNP haplotypes. While the reported method was verified with only a limited data set (N = 479) from Brown Swiss, Guernsey, Holstein, and Jersey cattle, some of the MS-SNP haplotype associations were concordant across these phylogenetically diverse breeds. This implied that some haplotypes predate modern breed formation and remain in strong linkage disequilibrium. To expand the utility of MS allele imputation across breeds, MS and SNP data from more than 8000 animals representing 39 breeds (Bos taurus and B. indicus) were used to predict 9410 SNP haplotypes, incorporating an average of 73 SNPs per haplotype, for which alleles from 12 MS markers could be accurately be imputed. Approximately 25% of the MS-SNP haplotypes were present in multiple breeds (N = 2 to 36 breeds). These shared haplotypes allowed for MS imputation in breeds that were not represented in the reference population with only a small increase in Mendelian inheritance inconsistancies. Our reported reference haplotypes can be used for any cattle breed and the reported methods can be applied to any species to aid the transition from MS to SNP genetic markers. While ~91% of the animals with imputed alleles for 12 MS markers had ≤1 Mendelian inheritance conflicts with their parents' reported MS genotypes, this figure was 96% for our reference animals, indicating potential errors in the reported MS genotypes. The workflow we suggest autocorrects for genotyping errors and rare haplotypes, by MS genotyping animals whose imputed MS alleles fail parentage verification, and then incorporating those animals into the reference dataset.

Termografia digital por infravermelho do escroto e qualidade do sêmen em touros nelore (bos taurus indicus)

Aimed to study the relationship between scrotal temperature on semen quality by use of infrared thermography in Nelore bulls bred extensively. There were scrotal thermography and semen collection. Climatic data were collected by means of globe thermometer. The thermograms of the scrotum were obtained by digital infrared thermography. We carried out ejaculates through electroejaculation for analysis of quantitative and qualitative characteristics. For the data, we used analysis of variance and subsequently applied the Tukey test at 5%. (p <0.05) between animals for sperm vigor, sperm motility between animals and between crops, sperm concentration, total sperm and total viable sperm between groups, between harvests for the scrotum temperatures and climate data. It was concluded that the surface temperatures of the scrotum and climatic factors influence the quality of semen. Thermography is recommended as a complementary test in the evaluation of reproductive bulls.

Detecção imunoistoquímica de receptores de estrógeno e progesterona no endométrio de vacas Nelore (Bos taurus indicus) durante o anestro pós-parto

Twenty-four postpartum anestrous Nelore purebred cows were used in the study. Anestrous was determined based on the reproductive history which was confirmed in each cow based on plasma progesterone concentration and by transrectal ultrasonography. Endometrial biopsies were collected. The animals were separated into two groups according to maximum follicular diameter- Group 1: follicles <6mm (n=12) and Group 2: follicles >-6mm follicles (n=12). The immunohistochemistry technique was employed to evaluate the presence of estrogen and progesterone receptors in the uterine glandular epithelium and stroma. High counts of positive nuclei and high intensity of immunostain for estrogen and progesterone receptors in the glandular epithelium and stroma were observed in the two groups. However, the immunostain intensity of progesterone receptors in the glandular epithelium was higher in Group 2 compared to Group 1. When glandular epithelium and stroma were compared within each group, the relative number of estrogen receptors in the Group 1 was higher in the glandular epithelium compared to stroma and the immunostain intensity for the progesterone receptor in Group 2 was higher in the glandular epithelium compared to stroma. The results suggest that the mechanisms that control the expression of endomerial receptors in the anestrus are similar to the ones observed during the estrus cycle.

Genome wide scan for quantitative trait loci affecting tick resistance in cattle (Bos taurus × Bos indicus)

Abstract Background In tropical countries, losses caused by bovine tick Rhipicephalus (Boophilus) microplus infestation have a tremendous economic impact on cattle production systems. Genetic variation between Bos taurus and Bos indicus to tick resistance and molecular biology tools might allow for the identification of molecular markers linked to resistance traits that could be used as an auxiliary tool in selection programs. The objective of this work was to identify QTL associated with tick resistance/susceptibility in a bovine F2 population derived from the Gyr (Bos indicus) × Holstein (Bos taurus) cross. Results Through a whole genome scan with microsatellite markers, we were able to map six genomic regions associated with bovine tick resistance. For most QTL, we have found that depending on the tick evaluation season (dry and rainy) different sets of genes could be involved in the resistance mechanism. We identified dry season specific QTL on BTA 2 and 10, rainy season specific QTL on BTA 5, 11 and 27. We also found a highly significant genome wide QTL for both dry and rainy seasons in the central region of BTA 23. Conclusions The experimental F2 population derived from Gyr × Holstein cross successfully allowed the identification of six highly significant QTL associated with tick resistance in cattle. QTL located on BTA 23 might be related with the bovine histocompatibility complex. Further investigation of these QTL will help to isolate candidate genes involved with tick resistance in cattle.

Patterns of cell proliferation and apoptosis by topographic region in normal Bos taurus vs. Bos indicus crossbreeds bovine placentae during pregnancy

Background: Placental and fetal growth requires high rates of cellular turnover and differentiation, which contributes to conceptus development. The trophoblast has unique properties and a wide range of metabolic, endocrine and angiogenic functions, but the proliferative profile of the bovine placenta characterized by flow cytometry analysis and its role in fetal development are currently uncharacterized. Complete understanding of placental apoptotic and proliferative rates may be relevant to development, especially if related to the pathogenesis of pregnancy losses and placental abnormalities. Methods: In this study, the proliferation activity and apoptosis in different regions of normal bovine placenta (central and boundary regions of placentomes, placentomal fusion, microplacentomes, and interplacentomal regions), from distinct gestation periods (Days 70 to 290 of pregnancy), were analyzed by flow cytometry. Results: Our results indicated that microplacentomes presented a lower number of apoptotic cells throughout pregnancy, with a higher proliferative activity by the end of gestation, suggesting that such structures do not contribute significantly to normal of placental functions and conceptus development during pregnancy. The placentome edges revealed a higher number of apoptotic cells from Day 170 on, which suggests that placentome detachment may well initiate in this region. Conclusion: Variations involving proliferation and apoptotic rates may influence placental maturation and detachment, compromising placental functions and leading to fetal stress, abnormalities in development and abortion, as frequently seen in bovine pregnancies from in vitro fertilization and cloning procedures. Our findings describing the pattern of cell proliferation and apoptosis in normal bovine pregnancies may be useful for unraveling some of the developmental deviations seen in nature and after in vitro embryo manipulations.

Novel Features of the Prenatal Horn Bud Development in Cattle (Bos taurus).

Whereas the genetic background of horn growth in cattle has been studied extensively, little is known about the morphological changes in the developing fetal horn bud. In this study we histologically analyzed the development of horn buds of bovine fetuses between ~70 and ~268 days of pregnancy and compared them with biopsies taken from the frontal skin of the same fetuses. In addition we compared the samples from the wild type (horned) fetuses with samples taken from the horn bud region of age-matched genetically hornless (polled) fetuses. In summary, the horn bud with multiple layers of vacuolated keratinocytes is histologically visible early in fetal life already at around day 70 of gestation and can be easily differentiated from the much thinner epidermis of the frontal skin. However, at the gestation day (gd) 212 the epidermis above the horn bud shows a similar morphology to the epidermis of the frontal skin and the outstanding layers of vacuolated keratinocytes have disappeared. Immature hair follicles are seen in the frontal skin at gd 115 whereas hair follicles below the horn bud are not present until gd 155. Interestingly, thick nerve bundles appear in the dermis below the horn bud at gd 115. These nerve fibers grow in size over time and are prominent shortly before birth. Prominent nerve bundles are not present in the frontal skin of wild type or in polled fetuses at any time, indicating that the horn bud is a very sensitive area. The samples from the horn bud region from polled fetuses are histologically equivalent to samples taken from the frontal skin in horned species. This is the first study that presents unique histological data on bovine prenatal horn bud differentiation at different developmental stages which creates knowledge for a better understanding of recent molecular findings.

The Cell Agglutination Agent, Phytohemagglutinin-L, Improves the Efficiency of Somatic Nuclear Transfer Cloning in Cattle (Bos taurus)

One of the several factors that contribute to the low efficiency of mammalian somatic cloning is poor fusion between the small somatic donor cell and the large recipient oocyte. This study was designed to test phytohemagglutinin (PHA) agglutination activity on fusion rate, and subsequent developmental potential of cloned bovine embryos. The toxicity of PHA was established by examining its effects on the development of parthenogenetic bovine oocytes treated with different doses (Experiment 1), and for different durations (Experiment 2). The effective dose and duration of PHA treatment (150 microg/mL, 20 min incubation) was selected and used to compare membrane fusion efficiency and embryo development following somatic cell nuclear transfer (Experiment 3). Cloning with somatic donor fibroblasts versus cumulus cells was also compared, both with and without PHA treatment (150 microg/mL, 20 min). Fusion rate of nuclear donor fibroblasts, after phytohemagglutinin treatment, was increased from 33 to 61% (P < 0.05), and from 59 to 88% (P < 0.05) with cumulus cell nuclear donors. The nuclear transfer (NT) efficiency per oocyte used was improved following PHA treatment, for both fibroblast (13% versus 22%) as well as cumulus cells (17% versus 34%; P < 0.05). The cloned embryos, both with and without PHA treatment, were subjected to vitrification and embryo transfer testing, and resulted in similar survival (approximately 90% hatching) and pregnancy rates (17-25%). Three calves were born following vitrification and embryo transfer of these embryos; two from the PHA-treated group, and one from non-PHA control group. We concluded that PHA treatment significantly improved the fusion efficiency of somatic NT in cattle, and therefore, increased the development of cloned blastocysts. Furthermore, within a determined range of dose and duration, PHA had no detrimental effect on embryo survival post-vitrification, nor on pregnancy or calving rates following embryo transfer.

A comparison of the excretion rate of endogenous purine derivatives in the urine of Bos indicus and Bos taurus steers

Estimates of microbial crude protein (MCP) production by ruminants, using a method based on the excretion of purine derivatives in urine, require an estimate of the excretion of endogenous purine derivatives (PD) by the animal. Current methods allocate a single value to all cattle. An experiment was carried out to compare the endogenous PD excretion in Bos taurus and high-content B. indicus ( hereafter, B. indicus) cattle. Five Holstein - Friesian ( B. taurus) and 5 Brahman (> 75% B. indicus) steers ( mean liveweight 326 +/- 3.0 kg) were used in a fasting study. Steers were fed a low-quality buffel grass (Cenchrus ciliaris; 59.4 g crude protein/kg dry matter) hay at estimated maintenance requirements for 19 days, after which hay intake was incrementally reduced for 2 days and the steers were fasted for 7 days. The excretion of PD in urine was measured daily for the last 6 days of the fasting period and the mean represented the daily endogenous PD excretion. Excretion of endogenous PD in the urine of B. indicus steers was less than half that of the B. taurus steers ( 190 mu mol/kg W-0.75. day v. 414 mu mol/kg W-0.75. day; combined s.e. 37.2 mu mol/kg W-0.75. day; P< 0.001). It was concluded that the use of a single value for endogenous PD excretion is inappropriate for use in MCP estimations and that subspecies-specific values would improve precision.

The tolerance of steers (Bos taurus) to sorghum ergot (Claviceps africana) in a feedlot during the cooler months in sub-tropical Queensland

Two experiments tested the tolerance of steers (Bos taurus) to sorghum ergot (Claviceps africana) during cooler months in south-east Queensland. Sorghum grain containing 2.8% ergot and 28 mg/kg ergot alkaloids (84% dihydroergosine, 10% dihydroelymoclavine, 6% festuclavine) was incorporated into feedlot rations. In a previous study in summer–autumn, ergot (1.1–4.4 mg alkaloids/kg ration) severely reduced performance in steers when the temperature–humidity index (THI; dry bulb temperature °C + 0.36 dew-point temperature °C + 41.2) was ~70, whereas a THI of ~79 was tolerated by steers fed ergot-free rations. Experiment 1 was conducted in winter–spring, with rations containing 0, 2.8, 5.6, 8.2 or 11.2 mg ergot alkaloids/kg ration. All ergot inclusions depressed feed intake (14% average reduction) and growth rate (34% average reduction), even when the weekly average daily THI was less than 65. Rectal temperatures were occasionally elevated in ergot-fed steers (P < 0.05), primarily when the THI exceeded ~65. All ergot inclusions depressed plasma prolactin concentrations in steers. Experiment 2 was predominantly carried out in winter, with weekly average daily THI <65 throughout the experiment. Rations containing 0, 0.28, 0.55 or 1.1 mg ergot alkaloids/kg were fed for 4 weeks but produced no significant effect on feed intakes and growth rates of steers. Alkaloid concentrations were then changed to 0, 2.1, 4.3 and 1.1 mg/kg, respectively. Subsequently, feed intakes declined by 17.5% (P < 0.05), and growth rates by 28% (P > 0.05) in the group receiving 4.3 mg/kg alkaloid, compared with Controls. Plasma prolactin concentrations were depressed, relative to the Controls, by dietary alkaloid inclusion greater than 1.1 mg/kg, with alkaloid intake of 4.3 mg/kg causing the greatest reduction (P < 0.05). Cattle performance in these studies shows steers can tolerate up to ~2 mg ergot alkaloid/kg (0.2% ergot) in feedlot rations under low THI conditions (< ~60–65), but previous findings indicate a much lower threshold will apply at higher THI (>65).

The tolerance of steers (Bos taurus) to sorghum ergot (Claviceps africana) in a feedlot during the cooler months in sub-tropical Queensland

Two experiments tested the tolerance of steers (Bos taurus) to sorghum ergot (Claviceps africana) during cooler months in south-east Queensland. Sorghum grain containing 2.8% ergot and 28 mg/kg ergot alkaloids (84% dihydroergosine, 10% dihydroelymoclavine, 6% festuclavine) was incorporated into feedlot rations. In a previous study in summer–autumn, ergot (1.1–4.4 mg alkaloids/kg ration) severely reduced performance in steers when the temperature–humidity index (THI; dry bulb temperature °C + 0.36 dew-point temperature °C + 41.2) was ~70, whereas a THI of ~79 was tolerated by steers fed ergot-free rations. Experiment 1 was conducted in winter–spring, with rations containing 0, 2.8, 5.6, 8.2 or 11.2 mg ergot alkaloids/kg ration. All ergot inclusions depressed feed intake (14% average reduction) and growth rate (34% average reduction), even when the weekly average daily THI was less than 65. Rectal temperatures were occasionally elevated in ergot-fed steers (P < 0.05), primarily when the THI exceeded ~65. All ergot inclusions depressed plasma prolactin concentrations in steers. Experiment 2 was predominantly carried out in winter, with weekly average daily THI <65 throughout the experiment. Rations containing 0, 0.28, 0.55 or 1.1 mg ergot alkaloids/kg were fed for 4 weeks but produced no significant effect on feed intakes and growth rates of steers. Alkaloid concentrations were then changed to 0, 2.1, 4.3 and 1.1 mg/kg, respectively. Subsequently, feed intakes declined by 17.5% (P < 0.05), and growth rates by 28% (P > 0.05) in the group receiving 4.3 mg/kg alkaloid, compared with Controls. Plasma prolactin concentrations were depressed, relative to the Controls, by dietary alkaloid inclusion greater than 1.1 mg/kg, with alkaloid intake of 4.3 mg/kg causing the greatest reduction (P < 0.05). Cattle performance in these studies shows steers can tolerate up to ~2 mg ergot alkaloid/kg (0.2% ergot) in feedlot rations under low THI conditions (< ~60–65), but previous findings indicate a much lower threshold will apply at higher THI (>65).

Estudo sobre a migração transuterina e a razão sexual de conceptos em bovinos da raça Nelore (Bos taurus indicus)

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, 2016.

Importance of sperm genotype (indicus versus taurus) for fertility and embryonic development at elevated temperatures

Heat stress has negative effects on bovine reproduction, particularly for European breeds (Bos taurus taurus) that are less thermotolerant than zebu cattle (Bos taurus indicus). Here, the evidence that spermatozoa and oocyte both contribute to early embryonic resistance to heat shock is demonstrated. In addition, the use of reproductive biotechnologies to improve bovine thermotolerance, are outlined by comparing data from taurus, indicus and crossbred genotypes. (c) 2005 Published by Elsevier B.V.