Characterization of Instrument Transformers under Realistic Conditions: Impact of Single and Combined Influence Quantities on Their Wideband Behavior.

Instrument transformers (ITs) play a key role in electrical power systems, facilitating the accurate monitoring and measurement of electrical quantities. They are essential for measurement, protection, and metering in transmission and distribution grids and accurately reducing the grid voltage and current for low-voltage input instrumentation. With the increase in renewable energy sources, electronic converters, and electric vehicles connected to power grids, ITs now face challenging distorted conditions that differ from the nominal ones. The study presented in this paper is a collaborative work between national metrology institutes and universities that analyzes IT performance in measuring distorted voltages and currents in medium-voltage grids under realistic conditions. Both current and voltage measuring transformers are examined, considering influence quantities like the temperature, mechanical vibration, burden, adjacent phases, and proximity effects. The study provides detailed insights into measurement setups and procedures, and it quantifies potential errors arising from IT behavior in measuring distorted signals in the presence of the various considered influence quantities and their combinations. The main findings reveal that the temperature has the most evident impact on the inductive voltage transformer performance, as well as the burden, causing significant changes in ratio error and phase displacement at the lower temperatures. As for low-power ITs, establishing a priori the effects of adjacent phases and proximity on the frequency responses of low-power ITs is a complex matter, because of their different characteristics and construction solutions.

How Instrument Transformers Influence Power Quality Measurements: A Proposal of Accuracy Verification Tests.

The integration of renewable energy sources on a large scale in the electrical energy distribution systems, as well as the widespread of non-linear loads, has led to a significant increase in power quality (PQ) disturbances. For this reason, PQ monitoring is also becoming a key task in medium voltage (MV) grids. The measurement of PQ at MV levels can only be performed using instrument transformers (ITs) to scale down the level of voltage and current to levels suitable for the input stage of PQ instruments. However, no international standards currently require the verification of the errors introduced by ITs in the measurement of PQ phenomena. Moreover, this issue is only partially addressed in the scientific literature, where papers dealing with specific and limited aspects of the problem can be found. For this reason, this paper aims to comprehensively assess the issue, proposing IT accuracy verification tests for different PQ parameters. First, a set of PQ phenomena relevant for IT testing is chosen, as well as the associated ranges of variation, based on a review of the enforced standards and the scientific literature. For each selected PQ phenomenon, possible performance indices and test waveforms are proposed. Finally, the proposed procedure is validated by applying it to the characterization of two different types of commercial voltage transformers.

Impact of DC Transient Disturbances on Harmonic Performance of Voltage Transformers for AC Railway Applications.

This paper analyzes the impact of typical DC transient events occurring in railway grids on the frequency performance of instrument transformers (ITs) installed onboard trains and in AC substations for power quality (PQ) applications. PQ monitoring in railway systems is an issue of great interest because it plays a key role in the improvement of energy efficiency. The measurement chain for the PQ measurements, at 15 kV at 16.7 Hz and 25 kV at 50/60 Hz, commonly includes ITs to scale the voltage to levels fitting the input of the measurement units. Nevertheless, the behavior of ITs in the presence of PQ phenomena represents an open issue from a normative point of view, even for those installed in conventional AC power supply systems. In this context, the paper presents a possible definition of DC transient disturbances test waveforms, a measurement procedure, and a setup to assess the impact of these disturbances on the harmonic performances of ITs for railway systems. Preliminary experimental tests carried out on two commercial ITs under wide ranges of variation for the amplitude and the time duration of DC disturbances show that, in some cases, the error introduced in harmonic measurements can exceed 100%.

Laboratory Production of Equine Embryos.

Assisted reproduction technologies (ART) are well developed in humans and cattle and are gaining momentum also in the equine industry because of the fact that the mare does not respond to superovulation but can donate large numbers of oocytes through ovum pick up (OPU). After collection, the oocytes can be fertilized by intracytoplasmic sperm injection (ICSI) using a variety of stallion semen samples, even of poor quality, and the resulting embryos can establish high pregnancy rates after cryopreservation and transfer. The discoveries that equine oocytes can be held at room temperature without loss of viability and that an increase in vitro maturation time can double the number of embryos produced are fueling the uptake of the OPU technique by several clinics that are shipping oocytes of their client's mares to specialized ICSI laboratories for embryo production and freezing. In this article, we present a retrospective analysis of 10 years of work at Avantea with a special focus on the last 3 years. Based on our data, an average production of 1.7 to 2 embryos per OPU-ICSI procedure can be obtained from warmblood donor mares with a pregnancy rate of 70% and a foaling rate in excess of 50%. OPU-ICSI offers the added value of freezing embryos that allows the development of embryo commercialization worldwide to the benefit of top horse breeders who are endorsing this technology as never before.

Transferability and inter-laboratory variability assessment of the in vitro bovine oocyte fertilization test.

The dramatic increase in the number of animals required for reproductive toxicity testing imposes the validation of alternative methods to reduce the use of laboratory animals. As we previously demonstrated for in vitro maturation test of bovine oocytes, the present study describes the transferability assessment and the inter-laboratory variability of an in vitro test able to identify chemical effects during the process of bovine oocyte fertilization. Eight chemicals with well-known toxic properties (benzo[a]pyrene, busulfan, cadmium chloride, cycloheximide, diethylstilbestrol, ketoconazole, methylacetoacetate, mifepristone/RU-486) were tested in two well-trained laboratories. The statistical analysis demonstrated no differences in the EC50 values for each chemical in within (inter-runs) and in between-laboratory variability of the proposed test. We therefore conclude that the bovine in vitro fertilization test could advance toward the validation process as alternative in vitro method and become part of an integrated testing strategy in order to predict chemical hazards on mammalian fertility.

Transferability and inter-laboratory variability assessment of the in vitro bovine oocyte maturation (IVM) test within ReProTect.

The new European chemicals policy for the Registration, Evaluation, Authorization and Restriction of Chemicals (REACH) will most probably impose a dramatic increase in the number of animals required for reproductive toxicity testing. For this purpose, the development and validation of alternative methods is urgently needed in order to reduce the use of laboratory animals. The present study describes the inter-laboratory variability and the transferability assessment of an in vitro test able to identify chemical effects during the process of oocyte maturation in a bovine model. The test was developed/optimised within ReProTect, an integrated research project funded by the European Union, joining together 35 partners with complementary expertise in reproductive toxicology. Eight chemicals with well-known toxic properties were tested (benzo[a]pyrene, busulfan, cadmium chloride, cycloheximide, diethylstilbestrol, ketoconazole, methylacetoacetate, mifepristone/RU-486 and DMSO as solvent) on the in vitro maturation (IVM) assay in two well-trained laboratories using the established Standard Operating Procedures. The statistical analysis demonstrated the concordance of results across the laboratories and the reproducibility of the test. We therefore conclude that the IVM test could advance toward the process of validation as alternative in vitro method that, in combination with additional in vitro tests, can become part of an integrated testing strategy in order to predict chemical hazards on mammalian fertility.

Development of an in vitro test battery for assessing chemical effects on bovine germ cells under the ReProTect umbrella.

Current European legislation for the registration and authorization of chemicals (REACH) will require a dramatic increase in the use of animals for reproductive toxicity testing. Since one objective of REACH is to use vertebrates only as last resort, the development and validation of alternative methods is urgently needed. For this purpose ReProTect, an integrated research project funded by the European Union, joining together 33 partners with complementary expertise in reproductive toxicology, was designed. The study presented here describes a battery of two tests developed within ReProTect. The objective of these tests is the detection of chemical effects during the processes of oocyte maturation and fertilisation in a bovine model. The corresponding toxicological endpoints are the reaching of metaphase II and the formation of the pronuclei respectively. Fifteen chemicals have been tested (Benzo[a]pyrene, Busulfan, Butylparaben, Cadmium Chloride, Carbendazim, Cycloheximide, Diethylstilbestrol, Genistein, Ionomycin, Ketoconazole, Lindane, Methylacetoacetate, Mifepristone, Nocodazole and DMSO as solvent) demonstrating high intra-laboratory reproducibility of the tests. Furthermore, the responses obtained in both tests, for several substances, had a good correlation with the available in vivo and in vitro data. These tests therefore, could predictably become part of an integrated testing strategy that combines the bovine models with additional in vitro tests, in order to predict chemical hazards on mammalian fertility.

Comparative aspects of somatic cell nuclear transfer with conventional and zona-free method in cattle, horse, pig and sheep.

Nuclear transfer (NT) is a complex procedure that requires considerable technical skills. Over the years attempts have been made to simplify the micromanipulations involved and to make the procedure more user-friendly. A significant step forwards has been the development of the zona-free NT methods. We have used zona-free NT with mechanical aspiration of the metaphase plate as a mean of enucleation, in a comparative approach with the conventional nuclear transfer zona-enclosed method in cattle, horse, sheep and pig. The absence of the zona considerably facilitates the enucleation step and significantly increases cell fusion success. On the other hand, the culture of zona-free NT embryos requires the embryos to be cultured individually or anyway separated from each other to avoid aggregation and also requires to prolong the in vitro culture up to the blastocyst stage before transfer. Blastocyst rate is equal or higher with zona-free method as compared to zona-enclosed method while survival after cryopreservation and development to term is comparable. In conclusion, our findings, together with published data, demonstrate that the zona-free system described in this paper can significantly increase the output of NT blastocysts over the conventional zona-enclosed system.

In vitro fertilisation with frozen-thawed bovine sperm sexed by flow cytometry and validated for accuracy by real-time PCR.

The methodologies used for cytometric sorting of fresh spermatozoa never allowed a clear resolution of sexual chromosomes of frozen-thawed semen. To devise a novel method for the production of bovine predefined sexed embryos using frozen-thawed semen, sorting efficiency of different protocols was studied using a new quantitative real-time PCR method to verify the purity of sexed semen. To this aim, after Percoll separation, frozen-thawed samples were stained at different temperatures and concentrations of Hoechst 33342 using a short-incubation time. The concentration of Hoechst 33342 of 500 mug/ml at a temperature of 37 degrees C provided good and stable fluorescence signals. Preventing the sperm clustering by adding 0.6% BSA in the 90% Percoll fraction led to X-bearing sperms purity of 91+/-2%. Thereafter, sorted sperms were used for in vitro fertilisation (IVF). Despite the lower cleavage rates reported in the sorted groups when compared with the control groups (40 vs 48%, P<0.01), blastocyst formation in the sorted and control groups was not different (27 vs 24% of the cleaved respectively). The PCR analysis of 30 blastocysts confirmed 26 embryos to be correctly sexed (87%). Transfer of two embryos per recipient into six synchronised heifers resulted in four pregnancies. Two abortions occurred at day 60, while two pregnancies went to term delivering two female calves. In conclusion, high purity and repeatability of sorting was obtained with frozen-thawed bull semen that was subsequently used for IVF giving rise to viable embryos and offspring. In addition, real-time PCR revealed to be an optimal support for these studies, providing a rapid and reliable estimation of flow cytometric efficiency.

Somatic cell nuclear transfer in horses: effect of oocyte morphology, embryo reconstruction method and donor cell type.

The objective of the present work was to investigate and clarify the factors affecting the efficiency of somatic cell nuclear transfer (NT) in the horse, including embryo reconstruction, in vitro culture to the blastocyst stage, embryo transfer, pregnancy monitoring and production of offspring. Matured oocytes, with zona pellucida or after zona removal, were fused to cumulus cells, granulosa cells, and fetal and adult fibroblasts, and fused couplets were cultured in vitro. Blastocyst development to Day 8 varied significantly among donor cells (from 1.3% to 16%, P < 0.05). In total, 137 nuclear transfer-embryos were transferred nonsurgically to 58 recipient mares. Pregnancy rate after transfer of NT-embryos derived from adult fibroblasts from three donor animals was 24.3% (9/37 mares transferred corresponding to 9/101 blastocysts transferred), while only 1/18 (5.6%) of NT-blastocysts derived from one fetal cell line gave rise to a pregnancy (corresponding to 1/33 blastocysts transferred). Overall, seven pregnancies were confirmed at 35 days, and two went to term delivering two live foals. One foal died 40 h after birth of acute septicemia while the other foal was healthy and is currently 2 months old. These results indicate that (a) the zona-free method allows high fusion rate and optimal use of equine oocytes, (b) different donor cell cultures have different abilities to support blastocyst development, (c) blastocyst formation rate does not correlate with pregnancy fate and (d) healthy offspring can be obtained by somatic cell nuclear transfer in the horse.

Influence of probe size on the measurement accuracy of non-uniform ELF magnetic fields.

The accuracy of extremely low frequency (ELF) magnetic field measurements can be sensitively varied by the meter probe size when the spatial distribution of the magnetic flux density is highly non-uniform, e.g. if the measurement point is close to the field source. The deviation between measured and actual field varies from point to point depending on the probe dimension and on the source configuration, once the other measurement conditions are fixed. The analysis of this effect is developed through a two-dimensional numerical model that enables the evaluation of the actual field value and of the corresponding probe indication. The error distribution, i.e. the deviation between actual and measured value, is computed for magnetic fields generated by industrial three-phase systems under both balanced and unbalanced supply conditions. The analysis shows how, for a given source, the error depends not simply on the distance from the source in relation to the probe size, but on its spatial distribution, which is complex and cannot be a priori predicted without using a computational tool such as the one proposed.

Pregnancy: a cloned horse born to its dam twin.

Several animal species, including sheep, mice, cattle, goats, rabbits, cats, pigs and, more recently, mules have been reproduced by somatic cell cloning, with the offspring being a genetic copy of the animal donor of the nuclear material used for transfer into an enucleated oocyte. Here we use this technology to clone an adult horse and show that it is possible to establish a viable, full-term pregnancy in which the surrogate mother is also the nuclear donor. The cloned offspring is therefore genetically identical to the mare who carried it, challenging the idea that maternal immunological recognition of fetal antigens influences the well-being of the fetus and the outcome of the pregnancy.

Embryonic and foetal development of bovine oocytes treated with a combination of butyrolactone I and roscovitine in an enriched medium prior to IVM and IVF.

Cattle oocytes were maintained at germinal vesicles (GV) stage for 24 hr using a combination of two specific and potent inhibitors of M-phase promoting factor (MPF) kinase activity, butyrolactone I (BL-I) and roscovitine (ROS). The media used for inhibition were (a) TCM-199 only and (b) TCM-199 supplemented with serum, hormones and growth factors. The effective doses of inhibitors were 6.25 microM BL-I and 12.5 microM ROS in medium (a) and 50 microM BL-I and 12.5 microM ROS in medium (b). After inhibition, about 90% of the oocytes resumed meiosis and reached the metaphase II (MII) stage during 24 hr of maturation. Following fertilisation the percentage of cleavage (D +2), compacted morula (D +6), blastocysts on D +7 and D +8 and the survival to freezing and thawing of grade 1 embryos frozen on D +7 were not different between the experimental treated groups and the control. In order to evaluate early foetal development, two groups of five grade 1 D +7 blastocysts derived from treated oocytes and two groups of five control embryos were transferred nonsurgically in four synchronised recipient heifers. On D +27, the recipients were slaughtered and the foetuses were recovered. In both groups, six foetuses developed out of the 10 embryos transferred. In conclusion, several supplements can be added to the prematuration medium of bovine oocytes without reducing the quality of inhibition but also without improving their subsequent developmental competence versus treated oocytes in TCM-199 only and versus untreated control. Furthermore, the prematuration step used in this study does not interfere with normal foetal development during the first stages of organogenesis.