Newsletter October 2019
By professor Poul Hyttel
There has been a wish for an EliteOva Newsletter giving regular updates on the project in order to support the common knowledge and team spirit. We have concluded an EliteOva Steering Group Meeting on October 24, and this is the perfect background for starting such a newsletter. This first newsletter is a bit more extensive than what will follow in order to give everybody the necessary background information.
The overall goals of EliteOva are to implement ultrasound-guided ovum pick up (OPU) as well as in vitro production (IVP) and genomic selection of embryos in Danish cattle breeding and to secure that the resulting embryos and calves are healthy and the technologies are embraced by the industry and society.
In the following, short updates on the status and further plans of each individual work package (WP) is presented.
The EliteOva partner Institut for Fødevare- og Ressourceøkonomi (IFRO) has the mission of investigating the industrial and societal readiness levels of the technologies included in EliteOva. They are at the moment conducting the investigation of the industrial readiness level including interviews with farmers and responses to extensive questionnaires. Approximately 200 completed questionnaires are expected to be ready for analyses at the end of 2019. Subsequently, IFRO initiate investigations of the societal readiness level with respect to the consumers.
The EliteOva partner Masterrind has the mission of testing the Embryo Trans Biotech (ETB) media series against an alternative production system. They have completed a series of comparisons of IVP based on slaughterhouse ovaries and have now proceeded into the next phase, where OPU/IVP will be implemented in the testing.
The EliteOva partners Trans Embryo Genetics (TEG), ETB and University of Copenhagen, Department of Veterinary and Animal Sciences (UCPH-DVAS) collaborate on the pipeline for OPU/IVP of embryos and establishment of pregnancies. The process includes OPU, which is performed at TEG in Tirsvad, IVP, which is performed either at ETB/UCPH-DVAS or at TEG, and embryo transfer and calving, which takes place at TEG. So far, 15 calves have been born from “Wave 1” to prove that this basic pipeline is functional. Subsequently, an additional IVP laboratory was established at TEG, and for “Wave 2” the IVP took place in this laboratory. During “Wave 2”, 31 pregnancies have been produced in this new laboratory setting, which has also proven functional, and calving is now ongoing. During “Wave 3”, where OPU is conducted at TEG and IVP at UCPH-DVAS, acquisition of embryonic biopsies has now been implemented in the pipeline. This process has, unfortunately, been delayed due to a change in postdoc at UCPH-DVAS, but the very well-qualified appointed postdoc, Vahid Najafzadeh, has now established a reliable and reproducible biopsy procedure that has already successfully resulted in 3 pregnancies after the first round of embryo transfers. Moreover, DNA from the biopsies has been amplified, shipped to Eurofins and resulted in excellent SNP call rates (all above the critical level of 0.8 and most above 0.98). A pipeline is now being established where the SNP data, received by the EliteOva partner Viking Genetics (VG), will be conveyed to UCPH-DVAS (call rates as measure of DNA quality) and TEG (NTM and other breeding information). “Wave 3” will continue with OPU/IVP during 2019 and 2020 with the aim of reaching a total of 100 pregnancies.
The next technological process to be implemented is cryopreservation of the biopsied embryos. Experimental trials are presently ongoing at ETB/UCPH-DVAS, where IVP based on slaughterhouse ovaries are conducted and two different methods for cryopreservation of biopsied embryos are tested: Conventional slow-freezing and vitrification. Embryos are monitored for recovery by culture, differential cell counts and apoptosis assays. Alternative cryo-experiments at TEG have been discussed and plans will be developed.
The EliteOva partners VG and Aarhus University (AU) have the missions of deriving breeding indices according to the Nordic Total Merit (NTM) from the biopsy SNP data, finding potential genomic markers for bull IVP performance, and implementing novel traits as e.g. methane-emission in the breeding indices. Due to the delay of acquisition of the embryonic biopsies mentioned in WP2, WP3 has also suffered a delay but has now been initiated and a postdoc is in place at AU. The goal of finding potential genomic markers for bull IVP performance has been extended by the project EliteSemen, for which additional funding has successfully been secured from the Milk Levy Foundation by Jan B-M Secher. This project is ongoing and will support WP3 with valuable data. The implementation of novel traits as e.g. feed-efficiency and reduced methane-emission in the selection of IVP embryos for transfer has been an important societal and environmental aspect of EliteOva. The development of SNP-related tools for implementing these aspects into the breeding indices have been and are being developed in parallel projects conducted by VG and AU. EliteOva, on the other hand, develops the platform where these novel indices can be applied on embryos resulting in reduced generation intervals and a significantly increased power of selection adding up to more efficient breeding progression. In the original deliveries of EliteOva, the implementation of novel traits as e.g. feed-efficiency and reduced methane-emission in the selection of IVP embryos has been scheduled for quarter (Q) 36. As EliteOva runs for 16 Qs, the full benefit of these novel solutions is not expected to materialize until after the conclusion of the project. Interestingly, there are indications from AU that SNPs related to feed-efficiency, which is closely linked to methane-emission, is expected to already be implemented in 2020 in a Saved Feed Index that will be calculated together with the NTM resulting in much earlier capitalization on this aspect.
The EliteOva partners Danish Technical University (DTU), Université Laval (Laval), Canada, and Melior Life Sciences (Melior), US, have the mission of evaluating the normality of the IVP embryos and calves with respect to their transcriptome and epigenetic signatures. A very well-qualified postdoc, Maria Belen Rabaglino, has been appointed at DTU. Embryos for these investigations are being produced in the experimental herd of a subcontractor, Estonian University of Life Sciences, Tartu, Estonia, where an extensive series of OPU/IVP and in vivo embryo collection experiments have been conducted over the summer and fall. The experiments have, unfortunately, due to unforeseen events been delayed, but they will be concluded primo December. The goal is to collect 16 days old IVP and in vivo embryos for the transcriptomic and epigenomic analyses. The embryos will be processed for studies of gene expression, DNA methylation and histone modifications.
Calves for these studies are likewise being produced at TEG where pregnancies are ongoing in order to produce 3 calves from OPU/IVP and 3 from in vivo embryos. One month after calving, these calves will be euthanized and samples will be collected from liver, brain and blood (and potentially other organs). These samples will be processed for studies of gene expression, DNA methylation and histone modifications at DTU, Laval and Melior. The goal is to establish the blood as a marker for the DNA methylation of the inner organs, and the further objective is, with this tool in hand, to examine blood DNA methylation in a larger number of OPU/IVP calves from the project. These investigations will give a molecular epigenetic signature of the normality of the IVP calves.
The EliteOva partner UCPH, Department of Veterinary Clinical Sciences (UCPH-DVCS) has the mission of investigating the clinical normality of the IVP calves. Blood and placenta samples have been obtained from the calves born in “Wave 1”. This sample collection was set on hold in “Wave 2” due to budgetary matters, but will be initialized again when calves materialize from “Wave 3” where the important biopsies have been implemented. Calves that die during or after calving are processed for autopsy, and so far one calf with malformations has been encountered, which is not alarming and considered within normal range. Due to the delay in the implementation of the embryonic biopsies, the production of calves for WP5 has also been delayed. Combined with the need in WP5 to follow the calves for 6 months after birth, this delay may call for an non-cost extension of the project in order to secure a time frame, within which all calves reach the age of 6 months and the collected data can be analyzed. Conclusion of this question will be taken at the next Steering Group Meeting.
EliteOva is an ambitious project including many partners and a diversity of goals. Half way through the project we have succeeded in implementing a set of advanced technologies into an embryo production pipeline that across geographical barriers efficiently results in calves on the ground. We have encountered certain delays in the project, but they have been tackled with the help and flexibility of EliteOva partners. We are now moving into a very exciting phase of the project, where the cumulative set of technologies including OPU, IVP, acquisition of biopsies, cryopreservation and embryo transfer will be applied, and where we will acquire molecular information on the embryo’s breeding indices as well as their epigenetic normality.
Finally, I would like to thank Project Manager Betina W Jensen and project leader trainee Jan B-M Secher for keeping meticulous track of the complex management of a project like this and all EliteOva partners for investing enthusiasm and hard working hours into the project.