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Our objective is to evaluate the effects of feeding rumen-protected Met (RPM) throughout the transition period and early lactation on the lipid profile of the preimplantation embryos and the endometrial tissue of Holstein cows. Treatments consisted of feeding a total mixed ration with top-dressed RPM (Smartamine® M, Adisseo, Alpharetta, GA, United States; MET; n = 11; RPM at a rate of 0.08% of DM: Lys:Met = 2.8:1) or not (CON; n = 9, Lys:Met = 3.5:1). Endometrial biopsies were performed at 15, 30, and 73 days in milk (DIM). Prior to the endometrial biopsy at 73 DIM, preimplantation embryos were harvested via flushing. Endometrial lipid profiles were analyzed using multiple reaction monitoring-profiling and lipid profiles of embryos were acquired using matrix assisted laser desorption/ionization mass spectrometry. Relative intensities levels were used for principal component analysis. Embryos from cows in MET had greater concentration of polyunsaturated lipids than embryos from cows in CON. The endometrial tissue samples from cows in MET had lesser concentrations of unsaturated and monounsaturated lipids at 15 DIM, and greater concentration of saturated, unsaturated (specifically diacylglycerol), and monounsaturated (primarily ceramides) lipids at 30 DIM than the endometrial tissue samples from cows in CON. In conclusion, feeding RPM during the transition period and early lactation altered specific lipid classes and lipid unsaturation level of preimplantation embryos and endometrial tissue. [ABSTRACT FROM AUTHOR]

In PubMed, it is possible to find more than 40,000 papers on embryo evaluation in various species. However, there is no consensus or gold standard method on how to assess their developmental potential. In assisted reproduction the evaluation "problem" is not only limited to embryos but involves the gametes as well. This manuscript provides an overview of some possible applications of label-free microscopy, in particular we describe the potential of the holographic microscopy in the IVF lab. We describe the positive aspects of several currently available microscopy label-free systems. In conclusion, we believe that a next generation of microscopy able to give objective markers for gamete and embryo quality is around the corner. • Label-free microscopy to evaluate live gametes and embryos. • Non-invasive microscopy systems. • Evaluation of oocyte's nuclei and sperm mass. • CT-scan of embryos with holographic microscopy. • Morphological markers on live specimens. [ABSTRACT FROM AUTHOR]

Introduction: In the past 20+ years, several studies of bovine embryo production showed how the ratio of male to female embryos changes if embryos are made in vivo or in vitro. It is known that in in vitro systems, the sex ratio is in favor of males when there are high levels of glucose, and favors females when the principal energetic substrate is one other than glucose, like citrate.Objectives: The aim of this study was to evaluate the embryo metabolism during three important periods of in vitro development: the early development (from day 1 until day 3), the middle of culture (day 3 until day 5), and later development (day 5 until day 7).Methods: To obtain this information we evaluated the spent medium from each time period by 1H NMR.Results: Our results confirm that embryo metabolism is different between sexes. The new information obtained by identifies markers that we can use to predict the embryo sex.Conclusion: These results open a new, non-invasive method to evaluate sex of the embryos before the transfer. In the first period of embryo culture, valine concentration is good indicator (66.7% accurate), while in the last phase of culture, pyruvate depletion is the best marker (64% accurate) to evaluate the sex of the embryo. [ABSTRACT FROM AUTHOR]