Microstructure of Holstein and Gir breed adult bovine hooves: histomorphometry, three-dimensional microtomography and microhardness test evaluation / Microestrutura de cascos de bovinos adultos das raças Holandesa e Gir: avaliação por histomorfometria, microtomografia tridimensional e teste de microdureza

This study aimed to characterize and compare the aspects involved in the microstructural formation of the Holstein and Gir breed hoof wall by histomorphometry, three-dimensional microtomography and microhardness test techniques. Seventy-two (18H/18G thoracic and 18H/18G pelvic) Holstein (H) and Gir (G) breed hooves were collected in slaughterhouses. The hooves were divided into six groups according to breed and age group: 24 to 36 months (C1H/C1G), 36 to 60 months (C2H/C2G) and over 60 months (C3H/C3G). The normality and homogeneity analysis of morphometric variables by the Kolmogrov-Sminov and Bartlett tests was conducted as a statistical model. Once the assumptions were met, the Sigmastat 3.5 software was used and the means were compared by T test. The 5% probability level is considered. When the assumptions were not met, the means were compared by the Mann-Whitney nonparametric test, considering the 5% probability level. When comparing the Holstein and Gir breeds, no differences were noticed between them as to the length of the dermal papillae; young animals showed thicker papillae than adult animals; the Holstein breed hooves showed higher amount of 7pores on the wall and on the sole compared to the Gir breed; Holstein cattle hooves showed greater microhardness than Gir cattle; there was no microhardness difference between pigmented and non-pigmented hooves of Holstein and Gir cattle.

O presente estudo objetivou caracterizar e comparar os aspectos envolvidos na formação microestrutural do estojo córneo de bovinos das raças Holandesa e Gir pelas técnicas de histomorfometria, microtomografia tridimensional e teste de microdureza. Foram coletados em frigoríficos, de forma igualitária entre as raças Holandesa (H) e Gir (G), 72 cascos (18H/18G torácicos e 18H/18G pélvicos). Os cascos foram divididos em seis grupos de acordo com a raça e a faixa etária: 24 a 36 meses, (C1H/C1G), 36 a 60 meses (C2H/C2G) e acima de 60 meses (C3H/C3G). Como modelo estatístico, realizou-se análise de normalidade e homogeneidade das variáveis morfométricas empregando testes Kolmogrov-Sminov e Bartlett. Atendidas as pressuposições, empregou-se o programa Sigmastat 3.5 e compararam-se as médias pelo teste T. Considerou-se o nível de 5% de probabilidade. Ao comparar as raças Holandesa e Gir, observou-se que não houve diferença entre elas quanto ao comprimento das papilas dérmicas. Os animais jovens apresentaram papilas mais espessas que os adultos. Os cascos da raça Holandesa apresentaram maior quantidade de poros na muralha e na sola em relação à raça Gir. Os cascos de bovinos Holandeses possuem microdureza maior que os de bovinos Gir. No estudo não se evidenciou diferença de microdureza entre cascos pigmentados e despigmentados de bovinos Holandeses e Gir.

Ovarian response to porcine FSH in association with ablation-induced or spontaneous follicular wave development during the estrous cycle in crossbred and Brazilian Warmblood mares.

The primary objective of this study was to examine the follicular and ovulatory responses following treatment with pFSH in association with ablation-induced or spontaneous follicular wave emergence or follicle deviation during diestrus in crossbred (Mangalarga×Arabian) and Brazilian Warmblood mares with a propensity for spontaneous multiple ovulations; secondary considerations were given to the collection of embryos. In Experiment 1, crossbred mares were administered (im) saline (control, n=7) or pFSH (25mg) when the largest follicle of the ablation-induced follicular wave reached ≥13mm (n=7) or ≥20mm (n=7) or, after pre-treatment ovulation (Day 0) on Day 6 (n=7). In Experiment 2, crossbred mares were administered (im) saline (control, n=10) or a larger dose of pFSH (50mg, n=7) when the largest follicle of the ablation-induced follicular wave reached ≥13mm. In Experiment 3, Brazilian Warmblood mares were administered (im) saline (control, n=7), pFSH (25mg, n=7 or 50mg, n=5) or EPE (12.5mg, n=7) as a positive control on Day 6. Ultrasonic technology was used to ablate all follicles ≥8mm and to monitor follicular development and detect ovulation. Treatment with pFSH or EPE was done twice daily until the largest follicle reached ≥32mm; thereafter, hCG (2500IU) was administered (iv) when the largest follicle reached ≥35mm. Artificial insemination was done 12h after hCG and embryo collections were done 8 d after post-treatment ovulations. In Experiments 1 and 2, treatment of crossbred mares with pFSH post-ablation in association with the expected time of wave emergence or follicle deviation did not (P>0.05) enhance the follicular or ovulatory responses or collection of embryos compared to controls. In Experiment 3, although the enhanced ovulatory response of mares to EPE at the expected time of spontaneous wave emergence was not different (P>0.05) from controls, it was greater (P<0.05) than the response to pFSH. In conclusion, the novelty of using follicle ablation prior to pFSH treatment at the time of wave emergence or follicle deviation did not enhance the follicular or ovulatory responses or collection of embryos to treatment in crossbred mares. In addition, the hypothesis that Brazilian Warmblood mares with a greater propensity for spontaneous multiple ovulations are as responsive to pFSH compared to EPE was not supported. Thus, the combined experimental results of the present study continue to support the general consensus that pFSH is relatively ineffective for follicular superstimulation/superovulation in mares.

Effects of timing of induced luteolysis in embryo donor mares on reproductive performance and pregnancy rate in recipient mares.

The objective was to evaluate the effects of giving prostaglandin F2(α) (PGF) to donor mares 48 h prior to embryo collection. Non-lactating donor mares (n = 20 estrous cycles in 10 mares), ranging from 2.5 to 10 y of age and 400 to 500 kg of body weight were used from September 2004 to February 2005 in the southern hemisphere (Brazil). Donor mares were randomly assigned in a cross-over design study. During a Treated cycle, 7.5 mg PGF was given 48 h prior to embryo collection, whereas in the Control cycle, 7.5 mg PGF was given at embryo collection. In Treated Cycles, serum progesterone concentrations decreased between the day of PGF treatment and the day of embryo collection (13.9 ± 5.4 and 0.5 ± 0.3 ng/mL, respectively; P < 0.05). In Treated versus Control cycles, the interovulatory interval was shorter (14.9 ± 0.9 vs 17.5 ± 1.1 d, P < 0.05). However, there was no significant difference between these groups for the interval from PGF to ovulation (average, 9.8 d), embryo recovery rate (average, 75%), embryo quality, uterine protein concentration, and pregnancy rate in recipient mares (average, 87% at 15 d after ovulation, with no pregnancy loss detected by 60 d). In conclusion, giving donor mares PGF 48 h prior to embryo collection reduced the average interovulatory interval by approximately 2.5 d, thereby potentially increasing the numbers of embryos that could be collected during a breeding season, with no deleterious effects on embryo recovery rate, embryo quality, or pregnancy rate in recipient mares.

Aspiration of equine oocytes from immature follicles after treatment with equine pituitary extract (EPE) alone or in combination with hCG.

This study examined the effect of treating mares with equine pituitary extract (EPE) alone or in combination with hCG on the recovery rate of immature follicles by transvaginal follicular aspiration (ovum pick-up; OPU). Ten normally cycling crossbred mares aged 3-15 years and weighing 350-400 kg were subjected to each of three treatments in a random sequence with each exposure to a new treatment separated by a rest cycle during which a spontaneous ovulation occurred. The treatments were (1) superovulated with 25mg EPE and treated with 2500 IU hCG, (2) superovulation with 25mg EPE, and (3) control (no exogenous treatment). Treatments 7 days after spontaneous ovulation; and all the follicles >10mm were aspirated 24h after the largest follicle achieved a diameter of 27-30 mm for control group, and most follicles reached 22-27 mm for the EPE alone treatment. To the group EPE+hCG, when the follicles reached 22-27 mm, hCG was administered, 24h before OPU. Superovulation increased the number of follicles available for aspiration. The total number of follicles available for aspiration was 61 in the EPE/hCG group, 63 in the EPE group and 42 in the control. The proportion of follicles aspirated varied from 63.5% to 73.8%. Oocyte recovery rate ranged from 15.0% to 16.7% and the proportion of mares that yielded at least one oocyte was 70% (7/10) in the EPE/hCG, 60% (6/10) in the EPE alone and 50% (5/10) in control group. The EPE/hCG treatment had a higher proportion of follicles with expanded granulose cells (64.4%) than the control (3.3%; p<0.05) and the EPE treatment (25.0%). The intervals from spontaneous ovulation to aspiration were similar for all treatments (11-12 days). However, superovulatory treatment significantly increased the aspiration to ovulation interval from 15+/-4 days for control to 27+/-15 days for EPE (p<0.05) and to 23+/-13 days for EPE/hCG treatment with commensurate increases in the time between spontaneous ovulations.

Use of a low dose of equine purified FSH to induce multiple ovulations in mares.

The effects of a low dose of equine purified FSH (eFSH) on incidence of multiple ovulations and embryo recovery rate in mares were studied. During the physiological breeding season in Brazil (19 degrees 45'45'S), 14 Mangalarga Marchador donor mares were used in a crossover study and another 25 mares of the same breed, between 3 years and 12 years of age were used as recipients for the embryo transfers. Donors were monitored during two consecutive oestrus cycles, an untreated control cycle followed by a treated cycle, when eFSH was administered. In both cycles, after an embryo collection attempt on day 8 post-ovulation all mares received 7.5 mg dinoprost and had their two largest follicles tracked daily by ultrasonography until the period of ovulation. Mares were inseminated every 48 h with extended fresh semen from a single stallion after the identification of a 35-mm follicle until the period of ovulation. Ovulations were induced by intravenous administration of 2.500 IU of human chorionic gonadotropin, upon detection of a 35- to 40-mm follicle. In the treated cycle, 5 mg eFSH was given intramuscularly once a day, from day 8 post previous ovulation until at least one follicle reached 35 mm in diameter. Embryo flushes were performed on day 8 of dioestrus (day 0 = ovulation). Treatment with eFSH resulted in higher (p < 0.05) ovulation rate and incidence of multiple ovulations compared to the control (1.6 vs 1.0 and 50% vs 0%, respectively--one mare had triple ovulation). However, embryo recovery rates in the control and treated cycles were similar (0.8 and 1.0, respectively; p > 0.05). Pregnancy rates in the recipient mares following embryo transfer were similar for the control and eFSH cycles (11/11 and 10/14, respectively). Additional studies are necessary in order to develop a low-dose protocol for the use of eFSH that brings a more consistent contribution to the efficiency of commercial equine embryo transfer programs.