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31.
The purpose of our work was to establish an efficient-oriented enucleation method to produce transgenic embryos with handmade cloning (HMC). After 41–42 h oocytes maturation, the oocytes were further cultured with or without 0.4 μg/ml demecolcine for 45 min [chemically assisted handmade enucleation (CAHE) group vs polar body (PB) oriented handmade enucleation (OHE) group respectively]. After removal of the cumulus cells and partial digestion of the zona pellucida, oocytes with visible extrusion cones and/or polar bodies attached to the surface were subjected to oriented bisection. Putative cytoplasts without extrusion cones or PB were selected as recipients. Two cytoplasts were electrofused with one transgenic fibroblasts expressing green fluorescent protein (GFP), while non-transgenic fibroblasts were used as controls. Reconstructed embryos were cultured in Well of Wells (WOWs) with porcine zygote medium 3 (PZM-3) after activation. Cleavage and blastocyst rates were registered on day 2 and day 7 of in vitro culture respectively. Meanwhile, the total blastocyst cell number was counted on day 7. We found that the difference was only observed between blastocyst rates (38.6 ± 2% vs 48.1 ± 3%) of cloned embryos with GFP transgenic fibroblast cells after CAHE vs OHE. With adjusted time-lapse for zonae-free cloned embryos cultured in WOWs with PZM-3, it was obvious that in vitro developmental competence after CAHE was compromised when compared with the OHE method. OHE enucleation method seems to be a potential superior alternative method used for somatic cell nuclear transfer (SCNT) with transgenic fibroblast cells.  相似文献   
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The aim of this work was to evaluate the incidence of potential ochratoxigenic mycoflora and ochratoxin A (OA) in poultry, pig and rabbit feeds. Eighty poultry, pig and rabbit feed samples were taken at random from factories located from Córdoba province, Argentina, over a period of 8 months. Isolation and quantitative enumeration of fungal propagules were done on DRBC and DG18 media. The predominant species wereA. candidus, A. flavus,A. terreus,A. parasiticus,P. implicatum P. minioluteum,P. crustosum andP. citrionigrum. The distribution of sectionNigri species varied according to the feedstuffs analysed. The frequency ofA. niger var.niger was noticeably high in poultry feed samples on DRBC medium. TheNigri section species was present at moderate mean colony counts (CFU/g) from three feeds. Mycotoxin analysis of these samples showed that OA was detected in 15%, 10% and 12% of pig, poultry and rabbit feed samples, respectively. The mean levels detected ranged between 15 and 25 ng/g from three feeds. The presence of ochratoxigenic species ofNigri section and OA in feeds indicates the risk of potential exposure of poultry, pigs and rabbits through the ingestion of feeds.  相似文献   
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Contents Bovine oocytes and embryos produced in vitro were studied to determine the mitochondrial pattern of distribution, segregation and activity using DIOC 6 and Jc-1 fluorescence. The highest fluorescence level observed in mature oocytes was taken as 100% activity and six activity levels were estimated as follows: (1) 0%, (2) 1-15%, (3) 16-30%, (4) 31-50%, (5) 51-75% and (6) 76-100%. Three patterns of mitochondrial distribution were found: (1) diffused throughout the cytoplasm in oocytes and embryos, (2) pericytoplasmic in oocytes and embryos, and (3) perinuclear only in embryos. The segregation of mitochondria in blastomeres showed two distinct patterns: (1) symmetrical with an even mitochondrial population, and (2) asymmetrical with different numbers of mitochondria in each blastomere. In immature oocytes, mitochondrial activity was very low and the distribution was diffuse or negligible, while in mature oocytes the activity was high and the distribution was diffuse or pericytoplasmic. Competent embryos up to the 16-cell stage showed intermediate levels of activity (16-50%) but activity decreased thereafter up to the blastocyst stage. Non-competent embryos showed low levels of activity (1-15%) at all stages. These results suggest that mitochondria might play an important role during early development and that a minimum threshold of activity regulates the potential competence for reaching the blastocyst stage.  相似文献   
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This research was carried out to determine the effects of shearing of different genotypes of male lambs (one from pure Karayaka stock and two from its crossbreed) in winter on bodyweight gain, feed consumtion, feed conversion efficiency, rectal temperature and carcass yield. A total of 21 lambs aged 8 months were allocated randomly to experimental groups according to a 2 × 3 factorial arrangement for shorn and unshorn animals. The lambs were fed grass hay (100 g/lamb/day) and commercial concentrate feed ad libitum during a 54‐day of fattening period. The lambs were shorn in the middle of the experiment (27 days after beginning the trial). The interaction between the shearing treatment × genotype of the studied parameters was found to be insignificant. The shearing process increased the rectal temperature (P < 0.01), hot carcass yield (P < 0.05) and dressing percentage (P < 0.05) and decreased the weight of the edible inner organs (P < 0.01). Some of these parameters were affected by the genotypes. It was concluded that shearing male lambs in the winter can have a beneficial effect on the hot carcass weight and dressing percentage without affecting fattening performance, and the performance of Karayaka lambs and its crossbred male lambs were similar under the conditions of the present study.  相似文献   
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Objective To determine whether dogs and cats are potential reservoirs of Ross River (RR) and Barmah Forest (BF) viruses
Method Young seronegative female dogs and cats were experimentally exposed to the viruses using Ochlerotatus vigilax (Skuse) mosquitoes.
Results Only one of the 10 dogs and one of the 10 cats exposed to RR developed neutralising antibody. None of the animals developed detectable viraemia or clinical signs. One dog and three cats exposed to BF developed neutralising antibody. In addition, a serological survey of sera obtained from domestic dogs and cats residing in the Brisbane region indicated that 23.7% and 1.3% of dogs, and 14% and 2% of cats, had neutralising antibodies to RR and BF respectively.
Conclusions Although dogs and cats are exposed naturally to these viruses, and can become infected, they are unlikely to be important urban reservoirs of either virus.  相似文献   
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Piezoelectric materials have many applications at cryogenic temperatures. However, the piezoelectric response below 10 kelvin is diminished, making the use of these materials somewhat marginal. Results are presented on strontium titanate (SrTiO3), which exhibits a rapidly increasing piezoelectric response with decreasing temperature below 50 kelvin; the magnitude of its response around 1 kelvin is comparable to that of the best materials at room temperature. This "giant" piezoelectric response may open the way for a broad class of applications including use in ultralow-temperature scanning microscopies and in a magnetic field-insensitive thermometer. These observations, and the possible divergence of the mechanical response to electric fields at even lower temperatures, may arise from an apparent quantum critical point at absolute zero.  相似文献   
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