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Equine IVP (OPU-ICSI) pregnancy from our lab!

Equine IVP (OPU-ICSI) pregnancy from our lab!

In July of 2023, we also started working with equine gametes in the ivf laboratory of Milkmen Ltd.-Embrió Ltd.’s ET Center in Paks.

We succeeded in making embryos in the first round as well, but our second attempt in September 2023 undoubtedly resulted in beautiful blastocysts. After vitrification and warming two embryos were transferred, both of them implanted as detected by ultrasonography on Day 8., October 2, 2023. As far as we know this is the first pregnancy from in vitro produced equine embryos in the Eastern half of Europe.

Next time we continue with direct transfer after vitrification.


Zsófia Vigh PhD, embryologist and the team, as well as the veterinary team

The laboratory background was provided by László Volman DVM, owner


The original date of publication of this post: 08.10.2023.


Photos: the 5 blastocysts cultured in the 2nd round, the two blastocysts resulting in pregnancy before vitrification, on 10.09.2023; one of them at 4.5 days; all oocytes of the donor mare after in vitro maturation

The first IVP (IVF) calf was born from the laboratory of Milkmen Ltd. – Embryo Ltd.

The first IVP (IVF) calf was born from the laboratory of Milkmen Ltd. – Embryo Ltd.

The first result of our new in vitro lab testing (see here:   https://embryo.hu/en/2022/10/20/ivf-embryos-in-our-lab/) which can be seen with the naked eye, was born at Dr. Gyula Gyulay’s herd in Martonvásár on 27.07.2023.

The Blonde’d Aquitaine heifer calf from the desired mating was born from a second grade IVP blastocyst, after fresh transfer.


A successful OPU-IVP (IVF) embryo program at Milkmen Ltd.’s lab

A successful OPU-IVP (IVF) embryo program at Milkmen Ltd.’s lab

After the IVF laboratory testing with slaughterhouse oocytes at the end of last year (which resulted 2 pregnancies), on February 28 we started a more hopeful and important activity, that is, oocyte collection from live animals. The procedure is called OPU, i.e. Ovum Pick-Up, aspiration of oocytes from visible follicles on the ovary through Transvaginal Ovum pick-up technique with the help of ultrasound scanner.

The donor heifers received minimal hormonal stimulation to recruit more number of follicles in to a growing pool from ovarian reserve. We got 17 oocytes out of them with the help of our colleagues Gyula Gyulay DVM and György Pénzes DVM. 7 were of first grade but we kept all 17 oocytes in IVM step. 

After 21-24 hours of maturation we fertilized the matured oocytes with conventional semen and also with sexed semen. We denuded them on the third day, and then cultured for 6 days under optimal environment. The conditions were right, as were the steps of the process, because on the 7th day after IVF: 4 blastocysts were “smiled at us” under the microscope considering IVF day as day ‘0’. The 2 first greade and 2 second grade, but properly developed Holstein embryos were also freshly transfered into the synchronized recipients.

One of the donor heifer was already superovulated in December with the standard amount of FSH hormone and the embryos were flushed out after 7 days (MOET). Then we collect 4 structures from her in vivo, of which only two were transferable embryos. The same heifer now gave 2 good embryos out of 5 oocytes using much less hormone.

So the method, which already works routinely in many places around the world, can also be effective here. Although all parts of the process still require some development on our part, but according to our plans, the program will be followed by many similar ones and will take the leading role in our reproductive biology programs.


2022 – our first full year

2022 – our first full year

End of the old year, beginning of the new year – for many, this period is about summing up. We did no different, curiously evaluating the first full year of the EMBRYO Transfer center – now in possession of a significant amount of data.

The most important result of our work, which can already be seen with the naked eye, is that since August 2021, a total of 124 Holstein-Friesian calves have been born from embryo transfers here at the Milkmen’s herd. They are mostly heifers, as we inseminated the donors with sexed semen in about 95% of the programs. The breeding value of all animals is not yet known, however, the average of the calves born from the transfers in 2021 is 2763 GTPI. There are among them that have been flushed due to their conformation traits and are less valuable in terms of their genetics, but there are many of heifers over 2800 and several extremely valuable over 2900 GTPI.

After our start-up year the year before last, we asked the question many times: how could we do it better? Based on our experience, our own and literature data, we have optimized our own production system at several points, especially our superovulation protocols and the laboratory processing of embryos. We keep both paper-based and electronic documentation of the programs down to the smallest detail, and we can draw conclusions from the amount of data that can be published soon. A 300x magnification photo documentation is also made of each produced embryo, which is also a great help for us in the evaluation of the programs, as well as for illustration and education. In addition to these, we have introduced new techniques from which we expect a significant improvement in the long term – preconditioning, splitting, IVF embryos.

Due to faster genetic progress, for our own purposes we flushed 90% of embryos from heifers only with sexed sperm. The average number of transferable embryos per wash was 6.2. This number is slightly higher than the European average for 2021 published by AETE (6) and the 2020 world average published by IETS (5.8), and well above the 2020 US average published by AETA, which is 4.8. The results published by the three organizations refer to the milking average – heifers and cows together. It is well known in the industry that more in vivo embryos can be collected from cows, as well as that more and better quality IVD embryos can still be produced with conventional (unsexed) semen. During the embryo programs mentioned above, only 19% in Europe and only 45% in the USA used sexed semen. Taking these factors into account, we were satisfied with the 7.2 average (heifer + cow) produced in the first semester.

Especially in the first half of the year we achieved a pregnancy rate of 46% by transfer 130 frozen embryos, far more transfered into recipient cows than into heifers. The pregnancy rates achieved with fresh embryos was somewhat weaker due to the fact that we also used many second and third grade embryos. With them, it is not possible to achieve the pregnancy rates of first class, but at least we give them a chance (by the way we transfered also many 2nd grade frozen embryos with reasonable results). After all, in certain cases, one calf can be more valuable than 10 others in total, for example, last year our heifer from ET with the highest genomic breeding value was also born from a third-grade fresh embryo, from. We thought we were on track.

In the second part of the year came unfortunately the new, weak feed (caused by the long drought period), which brought with it, among other things, a decrease in embryo production and, above all, in PR%. But rather, we focus on the fact that, in the meantime, several positive things started/happened, which give impetus to our further work and take us forward on the road:

– We have produced A2A2 embryos for other herd, which have been transfered with a PR of over 40%. We also started working with Brown Swiss donors.

– Calves were born without zona pellucida (these ones are so difficult to find and identify in the dish that whoever succeeds deserves 2/2) and from preconditioned embryos.

– We transferred splitted, biopsied and IVF embryos. Cattle IVP (in vitro embryo production) can work here, we are now working on stabilizing the system.

– In the near future, we will know the genomic breeding value of the majority of embryos before transfer, thanks to our ongoing research project.

– And finally, but by no means least: our donor stable is ready behind the lab. In this way, we can receive heifers for embryo production, taking the burden of ultrasound examinations, superovulation and insemination off the owner’s shoulders. We also moved our own donors and recipients here, where it is even possible to feed them individually.

– We started the development of several educational and research collaborations with certain institutions, and the introduction of a new technology, for which we unexpectedly received enormous help.


All in all (besides the calves), we have gained a lot of experience and data in the past year, which, supplemented with the previous ones, we can make available to interested breeders/vets/students more effectively. Embryo production, sale, transfer, education. Our doors are still open for professionals, whether you want to get information or learn, you can do so here, we look forward to welcoming you.


The main picture shows the embryos produced in February by our donor Janka 8570. 3 calves were born from the flush (one from a preconditioned-frozen embryos) with genomic breeding values: 2825, 2933 and 2976 GTPI.


IVF embryos in our lab!

IVF embryos in our lab!

We started to running the smallest but most important new unit of the Milkmen’s EMBRYO Transfer Center: the in vitro lab. The first round of testing went with oocytes from slaughterhouse ovaries, which were fertilized on October 12, 2022. Seven days later, 6 Holstein-Friesian and Blonde d’Aquitaine embryos were transfered into the synchronized recipients.

The successful first program will be followed by a similar one before we start embryo production with OPU-IVF in our own herd and make it routine.

On behalf of Embriobos Ltd., we thank the contribution of dr. Gyula Gyulay!

Calves from preconditioned in vivo embryos

Calves from preconditioned in vivo embryos

At Milkmen Ltd. in Paks-Földespuszta, the first two calves were born from in vivo embryos preconditioned before freezing.

Our team has introduced the PTAT cell preconditioning technology in the processes of embryo cryopreservation and in vivo embryo production.

PTAT (pressure triggered activation of tolerance) technology applies a defined, cell specific mild treatment for the reproductive cells. The short treatment slightly alters the environmental conditions for a predetermined time. As a consequence cells react with the fundamental, coded cellular defence mechanism that strengthens them. Consequently treated cells retain their normal function better, perform better, resulting in better embryo survival, better pregnancy rate and better offspring. The technology is provided by Applied Cell Technology Ltd (http://appliedcelltechnology.com).

In our Embryo Transfer Center, on 05/11/2021, the 8791 heifer from our Vilma cow family gave 10 transferable quality embryos. The mating bull was Elon. Four of the collected embryos – three first and one second grade – are treated before freezing using the method described above (PTAT – pressure triggered learning of tolerance). The embryos were transfered into recipient heifers on 11. 26 and the two calves were born on August 15 and 24, 2022.

We used the equipment manufactured by Applied Cell Technology for the first time in this lab, and since then we have continuously incorporated it into the daily routine of our embryo programs. We assign a serious role to it in terms of increasing efficiency and publishing our research results.

Photo of the embryos and calves can be seen under this post.