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11.
Vesicles Cytoplasmic Injection: An Efficient Technique to Produce Porcine Transgene‐Expressing Embryos 
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下载免费PDF全文 CG Luchetti RJ Bevacqua MS Lorenzo MF Tello M Willis CP Buemo DM Lombardo DF Salamone 《Reproduction in domestic animals》2016,51(4):501-508
The use of vesicles co‐incubated with plasmids showed to improve the efficiency of cytoplasmic injection of transgenes in cattle. Here, this technique was tested as a simplified alternative for transgenes delivery in porcine zygotes. To this aim, cytoplasmic injection of the plasmid alone was compared to the injection with plasmids co‐incubated with vesicles both in diploid parthenogenic and IVF zygotes. The plasmid pcx‐egfp was injected circular (CP) at 3, 30 and 300 ng/μl and linear (LP) at 30 ng/μl. The experimental groups using parthenogenetic zygotes were as follows: CP naked at 3 ng/μl (N = 105), 30 ng/μl (N = 95) and 300 ng/μl (N = 65); Sham (N = 105); control not injected (N = 223); LP naked at 30 ng/μl (N = 78); LP vesicles (N = 115) and Sham vesicles (N = 59). For IVF zygotes: LP naked (N = 44) LP vesicles (N = 94), Sham (N = 59) and control (N = 79). Cleavage, blastocyst and GFP+ rates were analysed by Fisher's test (p < 0.05). The parthenogenic CP naked group showed lower cleavage respect to control (p < 0.05). The highest concentration of plasmids to allow development to blastocyst stage was 30 ng/μl. There were no differences in DNA fragmentation between groups. The parthenogenic LP naked group resulted in high GFP rates (46%) and also allowed the production of GFP blastocysts (33%). The cytoplasmic injection with LP vesicles into parthenogenic zygotes allowed 100% GFP blastocysts. Injected IVF showed higher cleavage rates than control (p < 0.05). In IVF zygotes, only the use of vesicles produced GFP blastocysts. The use of vesicles co‐incubated with plasmids improves the transgene expression efficiency for cytoplasmic injection in porcine zygotes and constitutes a simple technique for easy delivery of plasmids. 相似文献
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The ICSI procedure is potentially of great value for felids, and it has not been extensively studied in these species. The objectives of this work were to determine the best conditions for ICSI in the domestic cat (DC) to generate interspecific embryos by injecting cheetah (Ch) and leopard (Leo) spermatozoa. Firstly, DC oocytes were matured with insulin–transferrin–selenium (ITS) or without it (MM) and cultured using atmospheric (21%) or low (5%) oxygen tension after ICSI. The group ITS‐5%O2 showed the highest blastocyst rate (p < 0.05), 20.9% vs 8.7%, 7% and 6.5%, for MM‐21%O2, MM‐5%O2 and ITS‐21%O2, respectively. The best conditions were used to generate the interspecific embryos, together with ionomycin activation (Io) after ICSI. Interspecific embryos resulted in high rates of blastocysts that were not positively affected by Io activation: 32.6% vs 21% for Ch and Ch‐Io, 9.8% vs 21% for Leo and Leo‐Io, and 20% vs 17.4% for DC and DC‐Io. We also evaluated DNA‐fragmented nuclei of experiment 1 and 2 blastocysts, using TUNEL assay. The fragmented nucleus proportion was higher in the ITS‐5%O2 group, 67.6%. Surprisingly, interspecific blastocysts showed the lowest fragmented nucleus proportion: 27% and 29.9% for Ch and Leo, respectively. We concluded that ITS and 5%O2 improve blastocyst formation in DC, although with a concomitant increase in DNA fragmentation. Most importantly, cheetah and leopard spermatozoa were able to generate blastocysts without artificial activation, which suggests that developmental capacity of wild felid spermatozoa can be evaluated by interspecific ICSI. This technique should be used to assist wild felid reproduction. 相似文献
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2003年10月,我县某养猪专业户自繁自养的9头70日龄仔猪,突然发生一种以头部水肿、共济失调、惊厥和麻痹为特征的疫病,经临床检查、剖检变化、实验室检验确诊为仔猪水肿病. 相似文献
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DF COOK 《Australian veterinary journal》1990,67(8):310-311
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A significant increase in milk production, averaging 164 litres per cow per lactation (a 4.8% increase), was seen after cows infected with gastrointestinal nematodes, paramphistomes and Fasciola hepatica were treated with broad-spectrum anthelmintics. Three hundred and ninety pairs of cows from eight herds with year-round calving were studied. One cow in each matched pair was given 7.5 mg/kg fenbendazole, 7.5 mg/kg levamisole hydrochloride and 15 mg/kg oxyclozanide in March, May and August of one year; the other cow in the pair received no anthelmintic. The number of nematode and trematode eggs was significantly decreased in the faeces of treated cows. 相似文献
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ST Rolim Filho HFL Ribeiro GMF de Camargo DF Cardoso RR Aspilcueta‐Borquis H Tonhati KB Nunes WG Vale EM Barbosa KC de Sousa 《Reproduction in domestic animals》2013,48(5):705-709
The osteopontin gene may influence the fertility of water buffaloes because it is a protein present in sperm. The aim of this work was to identify polymorphisms in this gene and associate them with fertility parameters of animals kept under extensive grazing. A total of 306 male buffaloes older than 18 months, from two farms, one in the state of Amapá and the other in the state of Pará, Brazil were used in the study. Seven SNPs were identified in the regions studied. The polymorphisms were in gene positions 1478, 1513 and 1611 in the region 5′upstrem and positions 6690, 6737, 6925 and 6952 in the region amplified in intron 5. The SNPs were associated with the traits, namely scrotal circumference, scrotal volume, sperm motility, sperm concentration and sperm pathology. There were significant SNPs (p < 0.05) for all the traits. The SNP 6690 was significant for scrotal circumference, sperm concentration, sperm motility and sperm pathology and the SNP 6737 for scrotal volume. The genotype AA of SNP 6690 presented the highest averages for scrotal circumference, sperm concentration and motility and the lowest total number of sperm pathologies. For the scrotal volume trait, the animals with the largest volume were correlated with the presence of the genotype GG of SNP 6737. These results indicate a significance of the osteopontin gene as it seems to exert a substantial influence on the semen production traits of male buffaloes. 相似文献
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Clones of aspen (Populus tremuloides Michx.) were identified that differ in biomass production in response to O(3) exposure. (14)Carbon tracer studies were used to determine if the differences in biomass response were linked to shifts in carbon allocation and carbon partitioning patterns. Rooted cuttings from three aspen Clones (216, O(3) tolerant; 271, intermediate; and 259, O(3) sensitive) were exposed to either charcoal-filtered air (CF) or an episodic, two-times-ambient O(3) profile (2x) in open-top chambers. Either recently mature or mature leaves were exposed to a 30-min (14)C pulse and returned to the treatment chambers for a 48-h chase period before harvest. Allocation of (14)C to different plant parts, partitioning of (14)C into various chemical fractions, and the concentration of various chemical fractions in plant tissue were determined. The percent of (14)C retained in recently mature source leaves was not affected by O(3) treatment, but that retained in mature source leaves was greater in O(3)-treated plants than in CF-treated plants. Carbon allocation from source leaves was affected by leaf position, season, clone and O(3) exposure. Recently mature source leaves of CF-treated plants translocated about equal percentages of (14)C acropetally to growing shoots and basipetally to stem and roots early in the season. When shoot growth ceased (August 16), most (14)C from all source leaves was translocated basipetally to stem and roots. At no time did mature source leaves allocate more than 6% of (14)C translocated within the plant to the shoot above. Ozone effects were most apparent late in the season. Ozone decreased the percent (14)C translocated from mature source leaves to roots and increased the percent (14)C translocated to the lower stem. In contrast, allocation from recently mature leaves to roots increased. Partitioning of (14)C among chemical fractions was affected by O(3) more in source leaves than in sink tissue. In source leaves, more (14)C was incorporated into the sugar, organic acid and lipids + pigments fractions, and less (14)C was incorporated into starch and protein fractions in O(3)-treated plants than in CF-treated plants. In addition, there were O(3) treatment interactions between leaf position and clones for (14)C incorporation into different chemical fractions. When photosynthetic data were used to convert percent (14)C transported to the total amount of carbon transported on a mass basis, it was found that carbon transport was controlled more by photosynthesis in the source leaves than proportional changes in allocation to the sinks. Ozone decreased the total amount of carbon translocated to all sink tissue in the O(3)-sensitive Clone 259 because of decreases in photosynthesis in both recently mature and mature source leaves. In contrast, O(3) had no effect on carbon transport from recently mature leaves to lower shoots of either Clone 216 or 271, had no significant effect on transport to roots of Clone 216, and increased transport to roots of Clone 271. The O(3)-induced increase in transport to roots of Clone 271 was the result of a compensatory increase in upper leaf photosynthesis and a relatively greater shift in the percent of carbon allocated to roots. In contrast to those of Clone 271, recently mature leaves of Clone 216 maintained similar photosynthetic rates and allocation patterns in both the CF and O(3) treatments. We conclude that Clone 271 was more tolerant to O(3) exposure than Clone 216 or 259. Tolerance to chronic O(3) exposure was directly related to maintenance of high photosynthetic rates in recently mature leaves and retention of lower leaves. 相似文献
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Leaf area, IBA concentration and age of stockplants were all found to be important factors for successful rooting for vegetative
propagation using single-node cuttings of the two main African mahogany species: Khaya anthotheca and K. ivorensis. Cuttings with leaf area of 30–50 cm2 had the best rooting percentage and cuttings with about 30 cm2 had the most number of roots per cutting in K. anthotheca. Khaya ivorensis, cuttings with 10–30 cm2 leaf area had the highest rooting percentage. Cuttings collected from 1-year-old stockplants recorded the highest rooting
percentage and largest number of roots per cutting. Cuttings from 3-year-old stockplant of the same seeds sources had the
lowest rooting suggesting aging negatively impacted rooting ability. The effect of auxin concentration, on rootability was
examined with cuttings of K. anthotheca. An IBA concentration of 0.8% was the best exogenous auxin concentration for percentage rooting, number of roots per cutting
and the length of the longest root per cutting. 相似文献
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Overexploitation of the African mahoganies, Khaya anthotheca and K. ivorensis, could lead to their commercial extinction in the near future. Efforts to establish plantations to sustain supply have been discouraged by the shoot borer Hypsipyla robusta Moore. We hypothesized that there is a shade level at which Hypsipyla attack and branching are reduced, but height growth is adequate. We report on the growth of these African mahoganies and Hypsipyla attack under three different forest canopy shade levels: open (55% open sky), medium shade (26% open), and deep shade (11% open). Hypsipyla attack on K. anthotheca was 85%, 11%, and 0% attack in the open, medium, and deep shade treatments, respectively. However, growth in medium and deep shade was slow. Mean tree height for K. anthotheca was 413.8, 126.2, and 54.5 cm in the open, medium and deep shade treatments, respectively. K. ivorensis showed similar trends. The reduced growth under these shade levels limit the use of this strategy for controlling Hypsipyla attack. 相似文献