Molecular Evaluation of Developmental Competence of Oocytes Collected In Vivo from Buffalo and Bovine Heifers during Winter and Summer

Buffaloes and bovines are polyestrous and seasonal or annual livestock, respectively, that show reduced fertility during heat stress. To investigate whether reduced fertility is related to oocyte competence in both species, immature oocytes from buffalo and bovine heifers were collected during winter and summer and subjected to molecular analyses. In each season, heifers of both species had their follicular wave emergence synchronized with a standard protocol (Ferreira et al., 2011). Before being subjected to ovum pick up (OPU), cutaneous (CT; degrees C) and rectal (RT; degrees C) temperatures and respiratory rate (RR; breaths/min) were measured. Oocytes' RNA was extracted to evaluate the expression of target genes related to mtDNA replication/transcription (PPARGC1A, TFAM and MT-CO1), apoptosis (BAX and BCL2) and HS (HSP90AA1 and HSPA1AB). ACTB, HIST1H2AG and GAPDH were initially chosen as housekeeping genes. In buffaloes, CT (35.0 +/- 0.4 vs 23.8 +/- 0.5), RT (38.7 +/- 0.1 vs 38.0 +/- 0) and RR (21.3 +/- 1.2 vs 15.4 +/- 1.1) were higher during summer than winter. However, in bovine heifers, RT (38.7 +/- 0.1 vs 38.6 +/- 0.1) and RR (44.8 +/- 1.5 vs 40.6 +/- 1.5) were similar in both seasons, while CT (31.6 +/- 0.3 vs 30.2 +/- 0.3) was increased during summer. Reduced expression of ACTB, HIST1H2AG and GAPDH was evidenced during summer, disqualifying them as housekeeping genes. Similarly, the expression of all target genes was reduced during summer in oocytes of both species. In summary, physiological responses to heat stress seem to be more intense in buffalo than bovine heifers. However, in both species, negative effects of heat stress upon oocyte quality occur at the molecular level and affects genes related to several biological functions.

Sources of omega-6 fatty acids do not alter the rumen degradation and transit of fibre from dairy cow diets

The objective of this study was to evaluate the behaviour of fibre in the digestive tract on the basis of the passage kinetics of forage and concentrate particles in cows fed different omega-6 fatty-acid sources. The scientific hypothesis of this study was that omega-6 fatty acids do not interfere with the digestion of fibre in the diets of dairy cows. Five primiparous lactating Holstein cows were used in the experiment. The experimental diets were: control (C), ground soyabean (GS), cottonseed (CS), soyabean oil (SO), calcium salts of fatty acids (CSFA). The global mean estimates for the parameters of passage rate (gamma) were 0.038 and 0.055 h(-1) for forage and concentrate, respectively. The only significant effect with respect to the passage rate was a high negative correlation between the concentrate passage rate and dry matter intake. There was less undegradable neutral detergent fibre (NDF) in treatments without added lipid. Dietary supplementation with lipid sources does not alter the kinetic parameters of roughage and concentrate particle passage or in vitro NDF degradation. Sources of omega-6 fatty acids do not alter the rumen degradation and transit of fibre.

Inducing ovulation early postpartum influences uterine health and fertility in dairy cows

The objective of the current study was to evaluate the effect of GnRH early postpartum on induction of ovulation, uterine health, and fertility in dairy cows. Holstein cows without a corpus luteum (CL) at 17 +/- 3 DIM were assigned randomly to receive i.m. GnRH (n = 245) at 17 +/- 3 and 20 +/- 3 DIM or remain as controls (n = 245). Ovaries were scanned by ultrasonography twice weekly totaling 4 examinations. Ovulation was characterized by the appearance of a CL >= 20 mm at any ultrasound or CL <20 mm in 2 consecutive examinations. Clinical and cytological endometritis were diagnosed at 35 DIM. Compared with control, GnRH increased ovulation up to 3.5 d after the last treatment (78.7 vs. 45.0%) and did not affect the prevalence of clinical endometritis (23.9 vs. 18.6%) or cytological endometritis (30.9 vs. 32.8%). Prevalence of clinical endometritis increased in cows that had calving problems (32.6 vs. 15.9%) and metritis (40.6 vs. 15.8%). Metritis increased prevalence of cytological endometritis (50.7 vs. 23.5%). Treatment with GnRH did not affect pregnancy per artificial insemination at 32 (37.6 vs. 38.6%) or 74 d after artificial insemination (35.0 vs. 31.5%), but reduced pregnancy loss (6.8 vs. 18.1%). No overall effect of GnRH treatment on hazard of pregnancy was observed; however, an interaction between GnRH treatment and ovulation showed that GnRH-treated cows that ovulated had increased hazard of pregnancy by 300 DIM compared with GnRH-treated and control cows that did not ovulate (hazard ratio = 2.0 and 1.3, respectively), but similar to control cows that ovulated (hazard ratio = 1.1). Gonadotropin-releasing hormone early postpartiim induced ovulation without affecting uterine health, but failed to improve pregnancy per artificial insemination or time to pregnancy, although it reduced pregnancy loss.

Comportamento ingestivo de vacas leiteiras alimentadas com silagem de milheto ou milho

The objective of this study was to evaluate the ingestive behavior of dairy cows fed corn and millet silage with a 5 mm particle size without inoculant, millet silage with a 5 mm particle size with inoculant, millet silage with a 20 mm particle size without inoculant, millet silage with a 20 mm particle size with inoculant. Five cows Holstein x Gir, with approximately 100 days of lactation at the beginning of the experiment and a mean body weight of 550 kg, producing an average of 15 kg of milk per day were used. The animals were arranged in a randomized 5 x 5 Latin square. The total feeding time, rumination time, chewing time, number of alimentary bolus, rumination time for bolus and number of chews was determined. The variables that make up the feeding behavior were not affected when the cows were fed different millet silages compared with corn silage. Cows fed corn silage and millet silage of 5 mm particle size had a higher intake of dry matter and neutral detergent fiber. A particle size between 5 and 20 mm, the presence or absence of inoculum and the type of silage with DM at 28%, does not affect the total feeding time, ruminating and resting of cows with an average production of 15.2 kg per day. The presence of the inoculant does not affect the intake of dry matter and neutral detergent fiber. Cows fed diets containing corn silage or pearl millet silage with a 5 mm particle size fed more DM.

Effects of supplementation of calcium salts of polyunsaturated fatty acids on serum concentrations of progesterone and insulin of pregnant dairy cows

Forty-five non-lactating, pregnant Holstein animals (18 heifers and 27 multiparous cows; BW = 561 +/- 114 kg; BCS = 2.9 +/- 0.3; days pregnant = 110 +/- 56 d) were stratified by initial BW and BCS, and randomly assigned to receive daily (as-fed basis) 0.50 kg of ground corn plus 0.22 kg of kaolin (CON), calcium salts of saturated fatty acids (SFA) or polyunsaturated fatty acids (PF) for 14 d. Blood samples were collected on days 0, 7 and 14, immediately prior to (0 h) and 3, 6, 9 and 12 h after feeding, to determine the serum concentrations of P 4 and insulin. No treatment effects were detected for serum concentrations of P 4 (5.52, 6.13 and 5.63 +/- 0.41 ng/mL for CON, SFA and PF, respectively). No treatment effects were detected for serum concentrations of insulin (11.5, 10.5 and 10.1 +/- 1.43 mu IU/mL for CON, SFA and PF, respectively). Heifers had greater serum concentrations of P 4 than multiparous cows (6.35 vs. 5.16 +/- 0.42 ng/mL), but lower serum concentrations of insulin (7.0 vs. 14.4 +/- 1.49 mu IU/mL). Feeding 0.22 kg of calcium salts of polyunsaturated fatty acids is not sufficient to increase the serum concentrations of P 4 and insulin of non-lactating, pregnant dairy cows.

Effects of excessive energy intake and supplementation with chromium propionate on insulin resistance parameters in nonlactating dairy cows

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

Increasing length of an estradiol and progesterone timed artificial insemination protocol decreases pregnancy losses in lactating dairy cows

Our hypothesis was that increasing the length of an estradiol and progesterone (P4) timed artificial insemination (TAI) protocol would improve pregnancy per artificial insemination (P/AI). Lactating Holstein cows (n = 759) yielding 31 +/- 0.30 kg of milk/d with a detectable corpus luteum (CL) at d - 11 were randomly assigned to receive TAI (d 0) following 1 of 2 treatments: (8d) d - 10 controlled internal drug release (CIDR) and 2.0 mg of estradiol benzoate, d - 3 = PGF(2 alpha) (25 mg of dinoprost tromethamine), d - 2 = CIDR removal and 1.0 mg of estradiol cypionate, d 0 = TAI; or (9d) d - 11 = CIDR and estradiol benzoate, d -4 = PGF(2 alpha), d -2 CIDR removal and estradiol cypionate, d 0 TAI. Cows were considered to have their estrous cycle synchronized in response to the protocol by the absence of a CL at artificial insemination (d 0) and presence of a CL on d 7. Pregnancy diagnoses were performed on d 32 and 60. The ovulatory follicle diameter at TAI (d 0) did not differ between treatments (14.7 +/- 0.39 vs. 15.0 +/- 0.40 mm for 8 and 9 d, respectively). The 9d cows tended to have greater P4 concentrations on d 7 in synchronized cows (3.14 +/- 0.18 ng/mL) than the 8d cows (3.05 +/- 0.18 ng/mL). Although the P/AI at d 32 [45 (175/385) vs. 43.9% (166/374) for 8d and 9d, respectively] and 60 [38.1 (150/385) vs. 40.4% (154/374) for 8d and 9d, respectively] was not different, the 9d cows had lower pregnancy losses [7.6% (12/166)] than 8d cows [14.7% (25/175)]. The cows in the 9d program were more likely to be detected in estrus [72.0% (269/374)] compared with 8d cows [62% (240/385)]. Expression of estrus improved synchronization [97.4 (489/501) vs. 81% (202/248)], P4 concentrations at d 7 (3.22 +/- 0.16 vs. 2.77 +/- 0.17 ng/mL), P/AI at d 32 [51.2 (252/489) vs. 39.4% (81/202)], P/AI at d 60 [46.3 (230/489) vs. 31.1% (66/202)], and decreased pregnancy loss [9.3 (22/252) vs. 19.8% (15/81)] compared with cows that did not show estrus, respectively. Cows not detected in estrus with small (<11 mm) or large follicles (>17 mm) had greater pregnancy loss; however, in cows detected in estrus, no effect of follicle diameter on pregnancy loss was observed. In conclusion, increasing the length of the protocol for TAI increased the percentage of cows detected in estrus and decreased pregnancy loss.

Artificial activation of bovine and equine oocytes with cycloheximide, roscovitine, strontium, or 6-dimethylaminopurine in low or high calcium concentrations

Knowledge on parthenogenetic activation of oocytes is important to improve the efficiency of nuclear transfer (NT) and intracytoplasmic sperm injection (ICSI) because artificial activation of oocyte (AOA) is an essential step to achieve embryo production. Although different procedures for AOA have been established, the efficiency of in vitro production of embryos remains low, especially in equines and Bos taurus bovines. In an attempt to improve the techniques of NT and ICSI in bovine and equine species, we tested different combinations of drugs that had different mechanisms of action for the parthenogenetic activation of oocytes in these animals. The oocytes were collected, in vitro matured for 24 to 30 h and activated artificially, in the presence of low or high concentrations of calcium, with combinations of calcium ionophore (ionomycin) with cycloheximide, roscovitine, strontium, or 6-dimethylaminopurine (6-DMAP). For assessment of activation rates, oocytes were stained with Hoechst 33342 and observed under an inverted microscope. We showed that all combinations of drugs were equally efficient in activating bovine oocytes, with the best results obtained when high concentrations of calcium were adopted. For equine oocytes, high concentrations of calcium were not beneficial for the parthenogenetic activation and the combination of ionomycin with either 6-DMAP or roscovitine was effective in inducing artificial activation of oocytes. We believe that our preliminary findings provide some clues for the development of a better AOA protocol to be used with these species.

Cardiovascular effects of a continuous rate infusion of lidocaine in calves anesthetized with xylazine, midazolam, ketamine and isoflurane

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

Acúmulo e desaparecimento de forragem e variações na estrutura de pastos de capim-xaraés submetidos a intensidades de pastejo em lotação intermitente

The aim of this study was to evaluate the effects of residual leaf area index (rLAI), years of evaluation and grazing cycles on the accumulation and disappearance of forage, and changes in the pasture structure of xaraes palisadegrass submitted to grazing intensities and intermittent stocking in two summers. The experiment was carried out at the Faculdade de Ciencias Agrarias e Veterinaria of Unesp, Campus of Jaboticabal, SP and the intensities of grazing were defined by four rLAI: 0.8, 1.3, 1.8 and 2.3. When the canopy intercepted 95% of incident light, the animals were placed on the pasture for grazing and kept until the rLAI target has been reached. Pastures were grazed by non-lactating Holstein cows (Bos Taurus Taurus L.), using the technique of mob-stocking. It was evaluated the canopy height, accumulation, disappearance and morphological components of forage and rest period. Residual leaf area index from 1.3 to 1.8 are the best in the accumulation and disappearance of forage and pasture structure. It is observed variability of the accumulation and disappearance of forage and pasture structure for years of assessment and grazing cycles within each year.

Evaluation of clinical safety and anthelmintic efficacy of aurixazole administed orally at 24 mg/kg in cattle

The current study evaluated, in vivo, the clinical safety and the anthelmintic efficacy of 24% aurixazole (24 mg/kg), administered orally, in bovines. Two experiments were conducted: the first one evaluating the clinical safety of 24% aurixazole (24 mg/kg) in cattle, and a second one evaluating the anthelmintic efficacy of aurixazole (24 mg/kg) against gastrointestinal nematodes on naturally infected cattle. Based on the results of clinical safety, no alterations on clinical and haematological signs and on the biochemical values obtained in animals treated orally with aurixazole 24 mg/kg were observed. Regarding the results of reduction or efficacy, obtained by eggs per gram of faeces (EPG) counts, the formulation of aurixazole reached values superior to 99% (arithmetic means) in all post-treatment dates. In two occasions, this formulation reached maximum efficacy (100%). Comparing these results with the reduction percentages obtained by EPG counts, it is possible to verify that the values obtained by all three formulations were compatible with the efficacy results. Aurixazole reached maximum efficacy (100%) against Haemonchus placei, Cooperia spatulata and Oesophagostomum radiatum. Against Cooperia punctata, this formulation reached an efficacy index of 99.99%. Regarding aurixazole, no specific trials were conducted on the field in order to evaluate the behaviour of this molecule against helminths that are resistant to other molecules, specially isolated levamisole and disophenolat: Due to this fact, future studies will be necessary to assess the effectiveness of aurixazole against strains of nematodes that are resistant to levamisole and disophenolat, but the results of clinical safety and efficacy described in this study allow us to conclude that the aurixazole molecule, concomitantly with other measures and orally administered formulations, can be another important tool in the control of nematodes parasitizing bovines. (C) 2014 Published by Elsevier Ltd.

Historic of therapeutic efficacy of albendazol sulphoxide administered in different routes, dosages and treatment schemes, against Taenia saginata in cattle experimentally infected

The present study aimed to notify the history of albendazole sulphoxide (ALB-SO) and albendazole (ALBZ) efficacy against Taenia saginata cysticercus (Cysticercus bovis) parasitizing experimentally infected bovines. A total of 11 efficacy trials were performed between the years of 2002 and 2010. In order to perform these trials, animals were individually inoculated with 2 x 104 eggs of T. saginata in each study's day zero (DO). For every trial, a positive control group (untreated infected animals) and a negative control group (animals that were neither infected nor treated) were used. ALB-SO or ALB were administered in the different dosages, in different days of treatments. In a last study with this formulation, this active principle was administered orally, mixed with the mineral supplement, on the 60th DPI, in a dosage of 30 mg/kg. In all trials, on the 100th DPI, all animals were euthanized and submitted to the sequenced slicing of 26 anatomical segments (fragments of approximately five millimeters) for the survey of T. saginata cysticercus. With the obtained results it is possible to verify that in the first trials, conducted in 2002, ALB-SO reached, independently of dosage and treatment scheme, efficacies superior to 98% (arithmetic means). The trials conducted in 2005 (2.5 mg/kg on the 30th, 60th, and 90th DPI) obtained values of efficacy all inferior to 60%. In 2008, the trials with 2.5 and 7.7 mg/kg demonstrated efficacy values inferior to 40%, for both dosages and treatment schemes (30th/60th/90th DPI and 60th DPI). When this formulation was administered orally on the dosage of 30 mg/kg on the 60th DPI, the efficacy against T. saginata cysticercus reached 88.28%. ALB administered orally showed efficacy values of 0.0%, 29.88% and 28.64% in the dosages of 5, 10 and 15 mg/kg, respectively, using the treatment schemes described above for each dosage. Based on the results of these trials, conducted in an eight year period (2002-2010) using the sequenced slicing method for evaluating the efficacy of the aforementioned formulations against T. saginata cysticercus, it is possible to observe that, amongst the few molecules used in the chemotherapic treatment against T. saginata larvae, ALB-SO, administered in varied routes, dosages and treatment schemes, the studies conducted in 2008, 2009, and 2010, have a low therapeutic efficacy against C bovis in Brazil, while ALBZ had insignificant efficacy values against T. saginata larvae parasitizing experimentally infected bovines. However, future studies using molecular biology will be necessary to assess whether the difference on the efficacy of the ALB-SO can be related to strain or another specific factor. (C) 2013 Elsevier Inc. All rights reserved.

Cana-de-açúcar hidrolisada para vacas em lactação

Pós-graduação em Zootecnia - FCAV

Diurnal behavior of dairy cows on alfalfa pastures supplemented with corn silage and concentrate

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

Precocidade sexual em bovinos Nelore avaliada por ultrassonografia testicular

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