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Cumulus-oocyte complexes from hormone-stimulated 3-4-week-old (n=43) and 6-7-week-old (n=12) prepubertal lambs were matured in vitro and incubated with unsorted, or X- or Y-spermatozoa separated with a high-speed cell sorter (SX MoFlo)frozen-thawed. Presumptive zygotes were then cultured to the blastocyst stage, and transferred to recipients fresh or after cryopreservation (frozen). Oocyte cleavage was higher (p <0.05) with unsorted (515/926, 55.6%) than X- or Y-spermatozoa (261/672, 38.8% and 229/651, 35.2%, respectively) and blastocyst formation (% zygotes) by Day 9 of in vitro culture was lower (p <0.05) for X- (102/261, 39.1%) than unsorted spermatozoa (249/515, 48.3%), but did not differ between Y-spermatozoa (103/229, 45.0%) and unsorted spermatozoa, or between X- and Y-spermatozoa (p >0.05). For fresh embryos, survival to term was 50.0% (3/6) for unsorted, 0.0% (0/6) for X- and 16.7% (1/6) for Y-spermatozoa-derived embryos (p >0.05), and for frozen embryos was 4.0% (2/50) for unsorted, 9.1% (2/22) for X- and 2.9% (1/34) Y-spermatozoa-derived embryos (p >0.05). Of the two lambs born from X-spermatozoa-derived embryos, one was female (50%), and from the two Y-spermatozoa-derived lambs, both were male (100%), demonstrating that lambs can be produced after the transfer of fresh and cryopreserved IVP embryos derived from prepubertal lamb oocytes and frozen-thawed sex-sorted sperm.  相似文献   
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The aim of this study was to validate an enzyme-linked immunosorbent assay (ELISA) for the measurement of progesterone (P4) in mares. Specifically, the objectives were as follows: (1) to determine the specificity and sensitivity of the ELISA test for determination of P4, (2) to measure the potential agreement between the 2 people performing the test, and 3) to evaluate the effect of time on the outcome. Ten mares were sampled on the day before ovulation (D-1), and on days 1 (D1), 3 (D3), and 5 (D5) following ovulation, during the reproductive season. While mares were cycling regularly, estrus was induced by the injection of 5 mg of prostaglandin (PGF2) and monitored starting on the 4th day by daily transrectal palpation and ultrasonography to determine the time of ovulation. Blood was collected and all samples (n=96) were assayed for P4 by a semiquantitative ELISA, by chemiluminescent immunoassay, and by radioimmunoassay (RIA). Based on the RIA, values of P4 on D-1, D1, D3, and D5 were significantly different (P < 0.0001) with mean and standard deviation(s) of 0.004, s = 0.52; 2.05, s = 2.58; 8.37, s = 4.17; and 12.76, s = 4.00 ng/mL respectively. The sensitivity and specificity of the semiquantitative assay were 94% and 95%, respectively for the lowest values of P4 (< 1.0 ng/mL). The value of kappa was 0.90 between 2 individuals performing the test. In conclusion, these results suggest that the semiquantitative test may be used reliably and economically to evaluate P4 levels in equine plasma in the clinical  相似文献   
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The suitability of a sea urchin population for commercial harvesting and culturing depends on the timing and duration of the spawning season. Pilot studies have shown that the New South Wales echinoids produce mature gonads for several months.Investigations were made into the influence of those exogenous factors which could be modified under aquacultural conditions.Temperature and food appeared to affect gonad (roe) development, while depth and density did not. The relationship between macro and micro changes in the gonads was established and methods of estimating stages of gametogensis were compared.  相似文献   
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Natural phytoplankton populations were cultured in outdoor continuous cultures using fish-farm effluents as the source of nutrients. The dilution rate was assumed to be the integrating factor of phytoplankton growth and biomass development (flux and stock). In this context, the combined effects of (i) dilution rates of the outdoor culture and (ii) ambient conditions were tested on phytoplankton growth, biomass and cycling of the major nutrient elements (C, N and P). Experiments were carried out in outdoor polyester tanks (0.7 m deep), homogenised by gentle aeration. Si/P ratio was balanced at around 5 in the inflow in order to induce diatom domination while maintaining high N and P assimilation by phytoplankton. Nutrient cycling was assessed through analyses of the different forms of particulate and dissolved nutrients in the inflow and the outflow. Culture dilution rates determined the longevity of the culture and the assimilation efficiency of nutrients. Dissolved phosphorus was the most limiting nutrient. The optimal dilution rate was approximately 0.5 day−1 at 10 °C and 1.5 day−1 at 20 °C with a mean diatom biomass of 9 μM P. Under these conditions, 80% of the dissolved nutrients provided to the tanks were transformed, a production of 8 g C m−2 day−1 and an assimilation rate of 0.3 g P m−2 day−1 were recorded. Assimilation by diatoms was the major pathway of nutrient cycling. During the experiment, a bottom sediment developed progressively and this also played an important role in denitrifying the excess dissolved nitrogen in the fish-farm effluent. However, the results showed that diatom biomass can collapse and we hypothesize that this was the consequence of an increase in cellular sinking rates due to cell aggregation under nutrient or light stress. Modelling approaches are needed in future research in order to determine optimal dilution rates taking into account phytoplankton growth rates, nutrient inputs and ambient conditions (e.g. light and temperature).  相似文献   
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An absorbed enzyme immunoassay (EIA) test for Johne's disease in cattle was developed in which absorption of cross-reacting antibodies occurred as a rapid reaction in solution rather than overnight with whole organisms and a subsequent centrifugation step. Total test time was reduced to less than 2 h with a minimum of manipulations. The test was evaluated in cattle herds from Johne's disease-endemic and Johne's disease-free regions of Australia. Specificity was 99.8%. Calculations of sensitivity were affected by the history of the herd under test. However, the EIA detected in excess of 80% of animals before onset of clinical disease and 65% of faecal shedders were EIA positive on, or before, first detection of Mycobacterium paratuberculosis in their faeces. The test should aid epidemiological studies and be a useful tool in the management and control of Johne's disease.  相似文献   
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