Search

To ensure that animal welfare requirements and phenotypic characteristics of the newly produced transgenic lines are not compromised, an evaluation of all individuals is necessary. This can be inferred by the analysis of the growth and reproduction parameters. The present study was designed to determine the impact of the insertion of human granulocyte-colony stimulating factor (hG-CSF) transgene on growth and reproductive characteristics in first-generation (F1) goats from two transgenic lines. Bodyweight (BW) development (BW at birth, mean BW gain before weaning, BW at weaning, mean BW gain after weaning, BW at puberty), as well as reproductive parameters (age at puberty, ejaculate volume, concentration, total sperm per ejaculate, massal motility, progressive individual motility, major and minor defects) were similar (P > 0.05) between transgenic (T) and non-transgenic (NT) goats. Significant (P < 0.05) differences in mean (±s.d.) white blood cell count were observed between T and NT in first day of life (174.6 ± 14.7 x 10³ and 15.0 ± 4.0 x 10³ cells/µL), and during (66.8 ± 21.1 X 10³ and 17.0 ± 4.6 x 10³ cells/µL) and after (36.6 ± 4.0 x 10³ and 15.5 ± 2.2 x 10³ cells/µL) suckling, even though hG-CSF has not been detected in blood serum in any analysis. Although other cell counts were occasionally higher in T animals, differential counts showed that this difference was mainly due to an increased number of neutrophils, which represents 84.6%, 67.2% and 56.8% of total white blood cell count respectively, in the three time periods. Kidney and liver biochemical analyses indicated that all goats were healthy. Thus, it is possible to assume that all animals are normal and had no deleterious effects on either growth or reproductive parameters by the presence of transgene or as a consequence of leukocyte profile alteration. [ABSTRACT FROM AUTHOR]

Contents Seventeen adult and cyclic Moxoto goats were synchronized using 60 mg MPA vaginal sponge for 11 days and 50 μg cloprostenol, 48 h before sponge removal, and superovulated with 120 mg pFSH i.m. in decreasing doses at 12 h intervals for three consecutive days. In seven goats, 0.2 IU/kg BW/day of long acting insulin was subcutaneously injected at same time as pFSH, and in the other five goats, the same dose of insulin was injected for three consecutive days starting 24 h after mating. Finally, five goats were supplemented with an oral dose of 80 ml/goat/day of propylene glycol continuously during the experiment. The animals were flushed at 7 days after mating and the embryos were classified based on International Embryo Transfer Society criteria. Blood samples were collected every 3 days for insulin assay. Administration of insulin raised the insulin levels of the goats (p < 0.05), whereas in the group treated with propylene glycol, insulin rate was different only between FSH treatment and after mating (p < 0.05). Similar rates of recovery for total (80.05 ± 9.78%) or transferable structures (61.03 ± 15.13%) were obtained. Treatment was not influenced (p > 0.05) by responsiveness to superovulation, which averaged 64%. By contrast, insulin treatments were shown to increase the number of embryos considered excellent with respect to goats supplemented with propylene glycol (p < 0.05). When insulin was given before mating, a strong relationship (r = 0. 90) (p < 0.05) between number of transferable embryo and ovulations was observed in the animals. In conclusion, superovulated goats treated with low doses of exogenous insulin resulted in an enhancement in embryo quality, which was related to changes in circulating insulin concentrations. [ABSTRACT FROM AUTHOR]

We investigated the influence of the phase of the estrous cycle on mechanical responses elicited in sheep cervix by potassium chloride (KCl), acetylcholine chloride (ACh), prostaglandin F2α (PGF2α) and prostaglandin E1 (PGE1). The cervix of adult ewes ( n = 48) were classified according to the presence or absence of corpora lutea (luteal or follicular phase, respectively). Muscle strips of the circular and longitudinal layers were prepared in an organ bath and coupled to an isometric force transducer. Concentration–response curves were obtained noncumulatively. KCl and ACh produced concentration-dependent contractions in all preparations in both phases of the estrous cycle. However, maximum effect, EC50 and slope values of KCl and ACh were not significantly different between muscle layers, as well as between the phases of the estrous cycle. The prostanoid, PGF2α, produced a significant reduction in the amplitude of spontaneous contractions for all preparations. The depressant effect of PGF2α on spontaneous contractions of circular smooth muscle was significantly greater during the follicular than the luteal phase, whilst the depressant effect of PGF2α on the longitudinal layer did not differ between phases of the estrous cycle. PGE1 significantly reduced the amplitude of spontaneous contractions on circular but not on longitudinal preparations. In conclusion, we have characterized with in vitro preparations of circular and longitudinal muscle layers of ewes during the follicular and luteal phases of the estrous cycle, the parameters of the K- and ACh-induced contractions on cervix and the efficacy of PGF2α and PGE1 on inhibition spontaneous contractile activity. [ABSTRACT FROM AUTHOR]

Contents The aim of this study was to determine the period of genital tubercle (GT) migration using ultrasonography in Morada Nova sheep foetuses (n = 117) from natural mating (NM) and frozen embryo transfer (ET) to determine the window when foetal sexing can be determined. The examinations were performed using transrectal ultrasonography with a dual-frequency linear transducer (6.0 and 8.0 MHz) from day 30–54 of pregnancy at 48-h intervals. The period of GT migration of foetuses produced by NM varied from 36 to 46 days of pregnancy, resulting in an average of 39.5 ± 2.9 days. For foetuses derived from ET, GT migration varied from 42 to 52 days of pregnancy with an average of 48.5 ± 3.3 days, being possible the GT of foetuses from ET start to migrate 96 h later even if they are of the same gender. Migration of the GT occurred earlier (p < 0.05) in foetuses produced by NM and sexing accuracy for triplet pregnancies (77.8%) was significantly inferior (p < 0.05) to single (100%) and twin (92.9%) pregnancies for foetuses derived by NM. The results allow one to conclude that foetal sexing can be done from the 50th day onwards in foetuses produced by NM and from the 55th day onwards in foetuses derived from ET, and that multiple pregnancies compromise the sexing accuracy by ultrasonography. [ABSTRACT FROM AUTHOR]

Contents Sixteen local adult goats were submitted for 9 weeks to 2.09 (high group) and 0.54 (low group) × dietary maintenance respectively. During the experimental period, goats were weighed, oestrus was detected and plasma insulin, urea, non-esterified fatty acids and progesterone concentrations were assessed. At the end of the experiment, ovarian small follicles population was studied by histological analysis. Final weight loss in low group was 18.37 ± 2.02%, whereas weight gain of high group was 13.84 ± 2.70%. Insulin and urea were lower in low group, while non-esterified fatty acids were significantly higher. A lower number of fasted goats was in oestrus or ovulated and an extended length of oestrus (p < 0.05) and a higher frequency of short or long cycles (p < 0.05) were also observed. Fed animals showed heavier ovaries (p < 0.01) and a lower number of primordial follicles (p < 0.05). In restricted goats a significant qualitative alteration of follicle classes involved in the initiation process of primordial pool was found. In this phase, granulosa thickness and oocyte size were the most affected (p < 0.01). However in small follicles beyond the primary stage no differences were found between the groups in either number or qualitative characteristics (p > 0.05). Collectively, these results indicate that opposite dietary intakes for a medium period induce a composite reproductive response in goats and can regulate the early onset of follicle growth. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the relation of gestational age with embryonic/fetal measurements in goats through the real-time ultrasound method. Ultrasonographic examinations began on the 25th day after first mating in eight Anglo-Nubian nulliparous goats, using a Falco 100 ultrasound scanner fitted to 6/8 MHz linear-array and 3.5 MHz convex probe. On recorded scanning images, the embryonic vesicle diameter (EVD) and crown-rump length (CRL), head length (HL), bi-parietal diameter (BPD), thoracic diameter (TD), umbilical cord diameter (UCD), long axis of heart (LAH), and short axis of heart (SAH) and placentomes of embryo/fetus were measured using computer software. The relationships between gestational age and each parameter of the ultrasonic measurements were expressed as straight-line equations and plotted as linear regression using Statistica 7.0 software, being statistically significant at a level of P< 0.05. Mean values were correlated to fetal age, and inverse regression equations were created for each parameter. Results obtained showed the following values: EVD (r = 0.94), CRL (r = 0.94), HL (r = 0.74), BPD (r = 0.70), TD (r = 0.88), UCD (r = 0.73), LAH (r = 0.77), SAH (r = 0.80), and placentome (r = 0.71). Additionally, embryonic size increased from the first detection of the embryo to the end of the study. In conclusion, the ultrasonographic technique is a reliable method to estimate gestational age through the embryonic/fetal measurement once its increase follows a linear regression standard. CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico - Brasil). [ABSTRACT FROM AUTHOR]

Laparoscopic oocyte recovery (LOR) is a valuable tool for obtaining oocytes for in vitroembryo production. When preceded by a treatment of ovarian stimulation, this technique produces an increase in the amount of oocytes recovered. However, a little information has been found to respect the effect of successive hormonal treatments on both oocyte quantity and quality. Therefore, the objectives of this study were to evaluate the ovarian response and quantitative and qualitative COC production. Five adult crossbred goats were hormonally treated with intravaginal sponges containing 60 mg of medroxyprogesterone acetate (MAP, Progespon, Syntex, Buenos Aires, Argentina) for 11 days. In the 8th day of progestagen treatment, 50 μg of prostaglandin F2α analogue (Ciosin, Coopers, São Paulo, Brazil) was administered by i.m. injection. At this time, ovarian stimulation was initiated by the administration of 120 mg pFSH (Folltropin-V, Vetrepharm, Canada) distributed in five decreasing doses (30/30, 20/20, 20 mg), at 12-h intervals. The animals were fasted for 24 h before the laparoscopic procedure, which was performed just after the sponge removal. A laparoscopic system connected to a 22-gauge needle (WTA, Watanabe, Brazil) and a vacuum pump (Biovacuum, Biocom, Brazil) providing 30 mm Hg was used. All follicles with a size larger than 2 mm present in both ovaries were counted and aspirated. The collection medium was TCM-199 supplemented with HEPES (10 mM), heparin (20 IU mL-1), and gentamicin (40 μg mL-1). The COCs were graded based on presence of cumulus cells and cytoplasm homogeneity (I to IV). The hormonal treatment and LOR procedures were repeated three times at 14-day intervals. Data were expressed in percentage or mean ± SEM. The differences were analyzed using ANOVA and Tukey''s or Fischer''s exact test when appropriate, with P 0.05) for the number of follicles obtained in each LOR session: 17.0 ± 3.91, 18.75 ± 2.59, and 18.0 ± 4.73, respectively. Repeated LOR procedures also did not affect (P> 0.05) the number of aspirated follicle (15.0 ± 3.92, 15.5 ± 2.33, and 16.0 ± 4.36), resulting from the three sessions, respectively. Average recovery rates were not statistically different (P> 0.05), resulting in 74.7%, 62.9%, and 64.6% between sessions. With respect to the percentage of viable COCs (GI and GII) were not observed statistical differences (P> 0.05), as verified the follow values at 1st to 3rd sessions: 76.79%, 84.62%, and 74.19%. In conclusion, three successive hormonal stimulation LOR procedures did not cause detrimental effects on quantitative and qualitative oocyte production, suggesting that this protocol can be used for programs of in vitrogoat embryo production. This study was supported the following Brazilian agencies: FINEP, CNPq, FUNCAP, and CAPES. [ABSTRACT FROM AUTHOR]

Reusing intravaginal devices represents an important alternative to reduce costs; however, this practice may increase disease transmission. The aim of this study was to evaluate the efficacy of reusing autoclaved intravaginal progesterone devices for oestrous synchronization in Toggenburg goats in the breeding season was studied. This study was done in March and April of 2009, in Piau, MG, Brazil (latitude 21°35′ and longitude 43°15′). Sixty-seven Toggenburg nulliparous (n=17; 35.3±5.4kg and 3.3±0.2) and pluriparous (n=50; 52.9±9.8kg and 3.4±0.3) goats were assigned according to weight and body condition score (BCS, 1 to 5 scale) into 3 treatments. Animals received new devices (n=25; 48.2±11.5kg and BCS 3.4±0.3) containing 0.33g of progesterone (Eazi-Breed CIDR®, Pfizer Animal Health, São Paulo, Brazil) or autoclaved (121°C, 1 atm, 15min) devices previously used for 6 days (n=23; 48.3±13.0kg and 3.5±0.3) or 12 days (n=22; 48.2±11.0kg and 3.4±0.3). All goats received 5mg dinoprost (Lutalyse®, Pfizer Animal Health) in the vulvar submucosa on the day of CIDR insertion (Day 0) and 200 IU eCG (Novormon 5000®, Sintex Indústria Bioquímica, Buenos Aires, Argentina) 1 day before CIDR removal, also in the vulvar submucosa. The CIDR were removed on day 6, and goats were bred twice daily with fertile bucks at oestrous onset and 24h later if they were still in oestrus. Parametric variables were analysed by ANOVA and SNK tests by the SAEG®program. Nonparametric variables were analysed using the chi-square test by the BioEstat®program. The results are described as mean±SD. Oestrous response and conception rates did not differ (P>0.05) among goats treated with the new devices (75.0; 54.2%) or those previously used for 6 (81.8; 50.0%) or 12 days (71.4; 47.6%), respectively. No differences were detected between nulliparous (82.3; 52.9%) and pluriparous (72.0; 50.0%) goats. The interval from device removal to oestrus and duration of oestrus were not different (P>0.05) among animals receiving a new device (39.3±15.8; 30.7±16.6h) or previously used devices for 6 (32.7±11.5; 31.8±7.3h) or 12 day (40.8±20.7; 32.8±13.2h) treatments, respectively, or between nulliparous (41.6±14.9; 30.9±13.9h) and pluriparous (35.6±16.8; 32.1±12.8h) goats. Since no differences were detected in the evaluated variables among goats receiving reused autoclaved devices or new ones, it can be suggested that the autoclaving process did not affect the efficiency of reusing intravaginal progesterone devices for oestrous synchronization in Toggenburg goats in the breeding season. Probably, P4 concentrations in goats receiving reused autoclaved devices reached at least minimum concentrations to promote oestrous response, since a non-treated group would not show oestrus in this level of synchronization as in goats in this study. This technique can be a simple and valuable tool to reduce sanitary risks of disease transmission without altering fertility in goats. CNPq, Pfizer Animal Health, Embrapa Goats, and Sheep Research, CAPES. [ABSTRACT FROM AUTHOR]

Oocyte IVM is a long process during which oocytes acquire their ability to support the stages of development in a stepwise manner, ultimately reaching activation of the embryonic genome. The overall success of this process can be affected by factors such as hormonal treatment for ovarian stimulation. Thus, the current study aims to evaluate the possible effects of the ovarian stimulatory protocols on the goat oocyte quality and IVM rate. Adult and cyclic Canindé goats were heat-synchronized by means of intravaginal sponges impregnated with 60 mg medroxyprogesterone acetate (MAP, Progespon, Syntex, Buenos Aires, Argentina) inserted for 11 days coupled with a luteolytic injection of 50 μg cloprostenol (Ciosin, Coopers, São Paulo, Brazil) in the 8th day of treatment. The ovarian stimulation was carried out using one of the following protocols: a) standard multi-doses (MD) with 120 mg pFSH (Folltropin-V, Vetrepharm, Canada) distributed in five injections (30/30; 20/20; 20 mg) at 12 h intervals (n= 18); b) three- doses (TD) with 120 mg pFSH administered in three injections (60; 40; 20 mg) at 24 h intervals (n= 17); c) one shot (OD) of 70 mg pFSH plus 200 IU of eCG (Novormon, Syntex) administered 36 h before sponge removal (n= 17). In MD andTD groups, the pFSH injections started in Day 8 of progestagen treatment. The follicles were aspirated just after the sponge removal using laparoscopic oocyte recovery (LOR). This procedure was performed with a 22-gauge needle and a vacuum pump at 30 mmHg. The collection medium was TCM-199 supplemented with HEPES (10 mM), heparin (20 IU mL-1), and gentamicin sulfate (40 μg mL-1). COCs were classified as grade I, II, III, or IV based on visual criteria (Baldassarre H et al.2003 Theriogenology 56, 831-839). Good quality oocytes (grade I and II) were incubated in TCM-199 supplemented with cysteamine (100 μM), EGF (10 ng mL-1) and gentamicin sulfate (40 μgm L-1) at 38.5°C in a humidified atmosphere with 5% CO2in air for 24 h. Oocyte maturation was assessed by the visualization of first polar body under inverted microscope. Data were expressed as percentages and analyzed using the Fischer''s exact test. No statistical differences among hormonal treatments (P> 0.05) were observed for the percentage of the good quality oocytes, with 70.4 ± 3.0% of COCs graded in I and II. The IVM rate inTD (31.4%) was statistically lower than MD (31.4% v.46.5%, P= 0.04) group. However, no significant differences (P= 0.89) were observed between OD (45.2%) and MD groups. Thus, current results indicate that oocyte production for IVM can be facilitated using ovarian stimulation with the one shot FSH/eCG regime without affecting meiotic competence. In summary, OD and MD treatments can be used for oocyte IVM in an embryo production programme in Canindé goats. This study was supported by the following Brazilian agencies: FINEP, CNPq, FUNCAP, and CAPES. [ABSTRACT FROM AUTHOR]