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Effect of Oil Overlay on Inhibition Potential of Roscovitine in Sheep Cumulus‐Oocyte Complexes
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LF Crocomo WC Marques Filho CMV Ulian NS Branchini DT Silva CL Ackermann FC Landim‐Alvarenga SD Bicudo 《Reproduction in domestic animals》2015,50(3):410-416
Inhibitors of cyclin‐dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus‐oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 μm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor‐free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (?) at 38.5°C and 5% CO2. At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/?) exhibited total expansion while in both Rosco groups (+/?) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/?). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression. 相似文献
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AM Batista WA Gomes CCD Carvalho PLJ Monteiro Jr FLM Silva FC Almeida PC Soares GF Carneiro MMP Guerra 《Reproduction in domestic animals》2014,49(3):476-480
The aim of this study was to evaluate the effect of leptin administration during superovulation on in vivo goat embryo production. Ten mature does were superovulated with 133 mg follicle‐stimulating hormone (FSH) i.m. in six descending doses at 12‐h intervals. The goats received 4.8 μg/kg human recombinant leptin s.c. (leptin group, n = 5) or phosphate‐buffered saline (PBS) (control group, n = 5) with the first and second FSH doses. The does were mated and subjected to embryo collection by transcervical technique 6 days later. The total number of cells per embryo and the number of cells with fragmented DNA were assessed in selected blastocysts by combining Hoechst 33342 and terminal dUTP nick‐end labelling (TUNEL) staining. Plasma concentrations of oestradiol (E2) and progesterone (P4) were determined by electrochemiluminescence from the day of FSH treatment, on the day of superovulatory oestrus and on the day before embryo collection. Compared with the control group, the does that received leptin had a higher number of transferable embryos (p < 0.005), fewer embryos classified as degenerated (p < 0.001) and fewer TUNEL‐positive cells/blastocyst (p < 0.001). The number of transferable embryos was positively correlated with E2 concentrations on day of oestrus (r = 0.562; p < 0.01) and P4 concentrations on the day of embryo collection (r = 0.912; p < 0.001). We concluded that in vivo leptin administration during FSH treatment improved embryo quality and affected ovarian steroidogenesis in superovulated goats. 相似文献
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OHO Eckhardt SMMK Martins ME Pinese FC Horta AC Rosseto MA Torres AFC De Andrade BBD Muro CT Marino PHM Rodrigues AS Moretti 《Reproduction in domestic animals》2014,49(6):964-969
The aim of this study was to evaluate the reproductive performance of three parities of gilts treated or not treated with gonadotropin to induce puberty. Sixty gilts received 600 IU of equine chorionic gonadotropin (eCG) followed by 2.5 mg of porcine luteinizing hormone (LH) 72 h later. Fifty‐nine other gilts were exposed only to a mature boar for 15 min twice daily. Artificial insemination (AI) was performed at 0, 12 and 24 h after the detection of oestrus, and gestation was confirmed by ultrasound after 35 days. Sows were inseminated at the first post‐weaning oestrus. The total numbers of piglets born, piglets born alive, stillborn, mummified foetuses, as well as pregnancy and farrowing rates were evaluated for each of the three parities. Culling rates, farrowing intervals and weaning‐to‐oestrous intervals (WEI) were also analysed. Mean age at puberty and oestrous manifestation were not significantly different between treatments (p = 0.0639; 179.20 ± 17.52 compared with 173.96 ± 16.94, 91.66% compared with 94.92%) across the experimental period. However, females that underwent puberty induction showed modest increases both in the number of total pigs born and in the number of piglets born alive. In conclusion, puberty induction through exogenous gonadotropin administration in field conditions did not induce a more concentrated first oestrous manifestation, but trended to a modest increase in the number of pigs born alive in the first parity and a reduced culling rate during the first gestation. 相似文献
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ARB HIGGS RT NORRIS FC BALDOCK NJ CAMPBELL S. KOH RB RICHARDS 《Australian veterinary journal》1996,74(3):215-220
Objective: To investigate control options for contagious ecthyma (scabby mouth) in Australian sheep exported live to the Middle East.
Design: Prevalence, vaccination and modelling studies.
Procedure: One hundred and forty weaner sheep (less than 1 year old) on each of 106 farms in Western Australia (WA) and 18 farm groups of adult wethers received at a WA commercial feedlot were examined for lesions of scabby mouth. Sheep on a total of 26 farms in 3 States were divided into treatment and control groups for the vaccination study. A simple deterministic compartmental model was developed to establish which parameters had the greater effect on disease prevalence.
Results: The proportion of farms with evidence of scabby mouth in weaner sheep was 23.6% and, on those farms with the disease, the overall prevalence was 6.1%. At the feedlot, 4 out of 18 farm groups had 5 or more sheep with lesions on arrival. The overall prevalence in the 4 diseased groups was 5.2%. Sheep vaccinated on farm before trucking to the feed-lot had a lower prevalence of scabby mouth at the end of simulated shipping than controls. The main determinant of scabby mouth prevalence was the proportion of sheep immune to the disease.
Conclusion: A program of vaccination for scabby mouth will reduce the prevalence of disease during live export. However, using current technology it is not possible to deliver shipments of sheep to the Middle East that are guaranteed completely free of scabby mouth. 相似文献
Design: Prevalence, vaccination and modelling studies.
Procedure: One hundred and forty weaner sheep (less than 1 year old) on each of 106 farms in Western Australia (WA) and 18 farm groups of adult wethers received at a WA commercial feedlot were examined for lesions of scabby mouth. Sheep on a total of 26 farms in 3 States were divided into treatment and control groups for the vaccination study. A simple deterministic compartmental model was developed to establish which parameters had the greater effect on disease prevalence.
Results: The proportion of farms with evidence of scabby mouth in weaner sheep was 23.6% and, on those farms with the disease, the overall prevalence was 6.1%. At the feedlot, 4 out of 18 farm groups had 5 or more sheep with lesions on arrival. The overall prevalence in the 4 diseased groups was 5.2%. Sheep vaccinated on farm before trucking to the feed-lot had a lower prevalence of scabby mouth at the end of simulated shipping than controls. The main determinant of scabby mouth prevalence was the proportion of sheep immune to the disease.
Conclusion: A program of vaccination for scabby mouth will reduce the prevalence of disease during live export. However, using current technology it is not possible to deliver shipments of sheep to the Middle East that are guaranteed completely free of scabby mouth. 相似文献
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Simões-Mattos L Mattos MR Teixeira MJ Oliveira-Lima JW Bevilaqua CM Prata-Júnior RC Holanda CM Rondon FC Bastos KM Coêlho ZC Coêlho IC Barral A Pompeu MM 《Veterinary parasitology》2005,127(3-4):199-208
Over the last few years, several cases of feline leishmaniasis (FL) with cutaneous and visceral forms have been reported around the world. Nonetheless, the real susceptibility of cats to infection with Leishmania spp. and the outcome of leishmaniasis in these animals are poorly understood. Experimental studies on feline models will contribute to the knowledge of natural FL. Thus, in order to determine the susceptibility of domestic cats (Felis catus) to experimental infection with Leishmania braziliensis, 13 stray cats were infected with 10(7) promastigotes by the intradermal route in the ear and nose simultaneously and followed up for 72 weeks. Soon after infection, the earliest indication of a lesion was a papule on the ear at 2 weeks post-infection (w.p.i.). The emergence of satellite papules around the primary lesion was observed about 4 w.p.i. Two weeks later these papules coalesced and formed a huge and irregular nodule. Thereafter, there was lesion dissemination to the external and marginal surface of the ipsilateral ear, and later to the contralateral ear. At 10 w.p.i., some nodules became ulcerated. Nose lesions presented a similar evolution. At both sites, the largest lesion sizes occurred at 10 w.p.i. and started to decrease 15 days later. Ear and nose nodules healed at 32 and 40 w.p.i., respectively. Specific L. braziliensis IgG antibody titers (optical density> or = 0.01 as positive result) were detected as early as 2 w.p.i. (0.09 +/- 0.02) in only three animals (23%), and all cats had positive titers at 20 w.p.i. (0.34 +/- 0.06). Only three animals (38%) continued to show positive serology at 72 w.p.i. (0.08 +/- 0.02). Up to that time, none of the cats had lesion recurrence. In a feline model of cutaneous leishmaniasis, it seems that there is no correlation between active lesions and positive serology. The implications of these data are discussed. 相似文献
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AISB Villaverde EG Fioratti FC Landim-Alvarenga JC de Carvalho Balieiro CM Melo FO Papa MD Lopes 《Reproduction in domestic animals》2009,44(S2):309-312
This study was aimed at assessing the changes in sperm motion patterns and the percentage of acrosome reaction (AR) in domestic cat semen after treatment with either ionomycin or progesterone (P4 ). Ten ejaculates were collected from five tomcats using an artificial vagina, and were diluted, centrifuged and resuspended in a capacitation medium. Samples were evaluated and divided into seven equal aliquots and, after 2 h at 25°C, were incubated for 30 min at 38°C in 5% CO2 and then analyzed. Computer-assisted sperm analysis and a combination of three fluorescent probes were used to assess sperm plasma, acrosomal membrane integrity and mitochondrial transmembrane potential. Thirty minutes after the start of incubation, P4 was added (10 μg/ml) to the P1 group. Groups P2 and P3 were supplemented with P4 (10 and 20 μg/ml, respectively) only after 2 h of incubation, and groups I1 and I2 were supplemented with ionomycin (4 and 8 μ m , respectively) 2 h after incubation. Group E was supplemented with ethanol (0.6%) at 2 h after incubation and group C received no supplementation. Ionomycin and P4 treatments led to a hyperactivation-like sperm motion and an increase (p < 0.05) in the percentage of AR. Although a higher (p < 0.05) percentage of AR was obtained in group I2 when compared with all P4 groups, a decrease (p < 0.05) in total and progressive motility was observed in I2 group. As I1 group was similar to I2 to induce AR without diminishing sperm motility, we can conclude that ionomycin at 4 μ m seems to be more suitable to trigger AR in domestic cat sperm. 相似文献
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Effects of concanavalin A on the progesterone production by bovine steroidogenic luteal cells in vitro
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FC Destro I Martin FDC Landim‐Alvarenga JCP Ferreira JL Pate 《Reproduction in domestic animals》2016,51(5):848-852
The aim of this study was to evaluate the effects of concanavalin A (CONA) on the progesterone (P4) production by bovine steroidogenic luteal cells (LCs) in vitro. Luteal cells were collected during the mid‐luteal stage (at 10–12 days following ovulation) and processed in the laboratory. Luteal cells were grown for 7 days in a humid atmosphere with 5% CO2, with or without 10% foetal bovine serum, and were subjected to the following treatments: control: no treatment; CONA (10 μg/ml); LH (100 μg/ml); CONA + LH; LH (100 μg/ml) + prostaglandin F2α (PGF2α) (10 ng/ml); CONA + LH + PGF2α. Samples of the culture media were collected on days 1 (D1) and 7 (D7) for P4 quantification. The cells were counted on D7 of culture. Differences between treatments were considered statistically significant at p < .05. Culture in the presence of CONA decreased the P4‐secreting capacity of LCs on D7 of culture, particularly in the absence of serum. The cell numbers did not change between treatments. 相似文献
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