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The objectives of this study were (a) to assess the ovulatory response and embryo production of Hungarian Merino ewes after superovulation, (b) to investigate the factors influencing the efficiency of embryo transfer (ET) in Hungarian Merino ewes, (c) to compare the results of two ovarian stimulation protocols (PMSG and PMSG+FSH treatment) in Hungarian Merino ewes, and (d) to study how superovulation, laparoscopic insemination and surgical embryo retrieval (ER) affect the subsequent reproduction of Hungarian Merino donor females after an ET programme. There was no significant difference between the ovarian stimulation protocols in the ratio of donor ewes responding to superovulation nor in the average number of corpora lutea. However, the number of transferable embryos recovered per donor ewe was higher in the PMSG+FSH group. The proportion of transferable embryos, unfertilised oocytes and degenerated embryos did not differ between the treatment protocols. The total pregnancy rate was 53.4% (179/335). Neither the developmental stage of the embryo nor the number of transferred embryos affect the implantation of embryos. However, the increased number of transferred embryos positively influenced the pregnancy rate. No difference was found in the pregnancy rate between synchronised and nonsynchronised groups of recipients. Thirty-six out of 45 donor ewes (80%) became pregnant within one year after the ET programme, indicating that ovarian stimulation and surgical ER did not affect adversely their reproduction.

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Animal experiments are very important for the development of new assisted reproductive techniques (ART) for use in human and animal reproductive medicine. Most technical aspects of reproductive manipulation of humans and animals are very similar, and many components of successful human ART used nowadays have been derived from animal studies. In this study we examined (1) the use of 'non-contact' laser for assisted hatching, (2) whether spindles in living mouse oocytes could safely be imaged/examined by polarisation microscope (polscope) and (3) the influence of environment (e.g. temperature, in vitro culture, etc.) on spindle detection/visualisation. The data of the study presented here show that (1) laser assisted hatching (AH) is a fast, very accurate and safe procedure without any harmful effect on embryo development and it can support very effectively the implantation of embryos, (2) the use of polscope facilitates the evaluation of oocyte quality and the selection of oocytes with spindle, (3) by monitoring the spindle position during intracytoplasmic sperm injection (ICSI), we can reduce spindle damage and increase the chance of fertilisation. Further studies are underway to test the hypothesised connection between spindle birefringence and developmental capacity of oocytes/embryos.

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Cryoinjuries are almost inevitable during the freezing of embryos. The present study examines the possibility of using high hydrostatic pressure to reduce substantially the freezing point of the embryo-holding solution, in order to preserve embryos at subzero temperatures, thus avoiding all the disadvantages of freezing. The pressure of 210 MPa lowers the phase transition temperature of water to -21°C. According to the results of this study, embryos can survive in high hydrostatic pressure environment at room temperature; the time embryos spend under pressure without significant loss in their survival could be lengthened by gradual decompression. Pressurisation at 0°C significantly reduced the survival capacity of the embryos; gradual decompression had no beneficial effect on survival at that stage. Based on the findings, the use of the phenomena is not applicable in this form, since pressure and low temperature together proved to be lethal to the embryos in these experiments. The application of hydrostatic pressure in embryo cryopreservation requires more detailed research, although the experience gained in this study can be applied usefully in different circumstances.

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The aim of this study was to determine the reliability of oestrus symptoms of the bitch (vulvar swelling, vaginal fluid, attraction of the male, teasing) and that of the most frequently used examination methods (vaginal smear, vaginoscopy, serum progesterone determination) which indicate the stage of the cycle and allow to predict the optimal day of mating (ODM). The determination of the cycle stage was based upon the number of days left from the day of examination to the day of parturition (DTP). Our results indicate that none of the above-mentioned symptoms and examination methods is reliable alone for the determination of DTP, and even the complex picture obtained from the integrated results of different methods will not enable an exact estimation. ODM can be predicted with a higher reliability, and 1-2 days before it, it can be detected very safely. Since most bitches are presented for timing to practitioners, a scheme was created based upon a scoring system, which gives a point value to each individual symptom and examination result, and judges the bitch according to the total amount of points.

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To check the importance of Cd-induced iron deficiency in Cd stress, symptoms of Cd stress were compared with those of iron deficiency or the combination of these two stresses. Poplar plants grown in hydroponics with Fe-EDTA (e) or Fe-citrate (c) up to four-leaf stage were treated for two weeks either by the withdrawal of iron (Fedef), or supplying 10 μM Cd(NO3)2 in the presence (Cad) or absence of an iron source (Fedef + Cad). Cadmium and iron content of leaves developing under the stress was in the order of cCad > eCad > cFedef + Cad and cCad ≈ eFedef ≈ cFedef + Cad < eCad < cFedef, respectively. Growth inhibition was much stronger in Cad than Fedef plants. The inhibitory effects on CO2 fixation, maximal and actual efficiency of PSII, chlorophyll synthesis, as well as the stimulation of the accumulation of violaxanthin cycle components and increase in non-photochemical quenching were the strongest in cFedef+Cad plants, otherwise these parameters changed parallel to the iron deficiency of leaves. Tendency of changes in thylakoid composition were similar under Cad treatments and strong iron deficiency: particularly PSI and LHCII decreased. Therefore, the development of the photosynthetic apparatus under Cd stress was mainly influenced by the Cd-induced strong iron deficiency, while leaf growth was affected primarily by the presence of Cd.

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Acta Veterinaria Hungarica
Authors: Margit Kulcsár, Gabriella Dankó, H. G. I. Magdy, J. Reiczigel, T. Forgach, Angella Proháczik, Carole Delavaud, K. Magyar, Y. Chilliard, L. Solti, and Gy. Huszenicza

Maternal plasma leptin is elevated in ewes during pregnancy. The authors studied whether there was any relation between maternal plasma leptin and insulin concentrations, the number of fetuses and the circulating and faecal levels of gestagens. At the end of the breeding season in January the ovarian activity of Prolific Merino ewes was induced/synchronised with gestagen + eCG treatment. Ewes were inseminated artificially (AI) by laparoscopy. Blood and faecal samples were collected before AI (day 0) and again 41, 81 and 101 days later. The plasma levels of leptin (pL), insulin and progesterone (pP4), and the faecal P4 metabolite (P4-met) content were determined. The day 0 level of pL was significantly higher in pregnant (n = 24) than in non-pregnant ewes (n = 32). By day 41 the pL of pregnant animals had doubled, it showed a further moderate increase on day 81, and decreased slightly thereafter. During pregnancy pP4 and faecal P4-met rose continuously and were positively correlated at all stages. The mean levels of pL and pP4 and the faecal content of P4-met were lower in ewes bearing single (n = 12) than in those with 2 (n = 6) or 3-5 fetuses (n = 6). Analysis of variance demonstrated significant differences according to the number of fetuses in the pL and pP4, but not in P4-met (p = 0.042, 0.044, and 0.051, respectively). Leptin showed positive correlation with insulin before the AI but not during pregnancy. On days 41 and 81 pL showed a slight positive correlation with P4 and P4-met, which decreased slightly by day 101. This study shows that although leptinaemia is affected by the number of fetuses and the level of P4, pregnancy stage is a more important regulator than these additional factors.

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