2011 A.E.T.E. NEWSLETTER N

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1 January 2011 A.E.T.E. NEWSLETTER N 34 Editor: Dimitrios Rizos TABLE OF CONTENTS President's letter Claire Ponsart Upcoming Events & Continuing Education Opportunities.. A Visual update of the last AETE Scientific Meeting..... Winner of the Student Competition Josine Beek, Ghent University, Belgium.. Workshop I - Kuopio Sep 2010 Embryo Cryopreservation: Status and Future Directions Catherine Guyader-Joly... European statistical data of bovine embryo transfer activity 2009 Sybrand Merton... A.E.T.E. Next Conference: Chester, United Kingdom, 9 th -10 th of September Dear Colleagues, Our society has held its 26 th annual meeting in Kuopio, a very pleasant venue in the beautiful Finland. The meeting was attended by over 130 delegates, originating from 22 European countries: more than 25 attendees were originating from Scandinavian countries, thus indicating the dynamism of embryo transfer activity in this region and the local implication of the local organising committee. The conference was organised in 17 oral presentations, 2 workshops and a poster session with 64 posters. A.E.T.E. Newsletter Dimitrios Rizos, Editor drizos@inia.es No.34 December 2010 A biannual published by the European Embryo Transfer Association Web Site: Letters to the Editor are welcomed. Please include name, address, telephone, FAX, and address The presentation of Dr Yvan Heyman as the recipient of the AETE Pioneer Award was a special moment of the meeting. Dr Yvan Heyman received the award due to his real outstanding work in the field of cloning and nuclear transfer. Jean-Paul 1

2 Renard reminded in the laudatory speech his pioneering experiments on cervical transfer, embryo bisection and freezing in the 80s. Our society also took benefit of its expertise, its wariness and simplicity as mentioned by JP Renard from 1995 to 2002, when he served the AETE as Secretary in the board. genotypes together with an economy of embryo transfers. Mel Dejarnette presented retrospective data regarding the use of sexed semen with an epidemiological approach: across all breeds and parities, 88% of all singleton births reported to be conceived from use of sexed semen were female offspring. The other invited lectures were given by Sabine Kölle, Patrice Humblot and Mel Dejarnette. Sabine Kölle gave new insights into transport of gametes and embryos along the oviduct, using an innovative method of videomicroscopy: it was remarkable to observe that the mature COC immediately and firmly attached to the oviductal epithelium, when entering the ampulla. Patrice Humblot described the potential contribution of embryo related biotechnologies to the genomic selection: in the near future, use of embryo genotyping should be a possible way to select early embryos presenting interesting As every year, four students were selected to participate in the Student Competition. The Board members judged the students on their abstract, poster and oral presentation. Finally, Josine Beek, working at the Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium was unanimously designed as winner of the student competition and obtained a prize of 750 euros from the AETE society for her following research: she inhibited serine proteases using a specific inhibitor and observed a negative influence on sperm motility and in vitro fertilization (ivf) in pigs. Both workshops were very practical orientated. The first workshop, co-ordinated by Catherine Joly focussed on cryopreservation of - 2 -

3 embryos: several ET teams from Europe showed that cryopreservation of biopsied embryo is an actual concern, probably in relationship with embryo genotyping. Labelling of straws was also discussed in relationship with IETS recommendations. The second workshop, coordinated by Jędrzej Jaskowski was dedicated to the non-invasive evaluation of embryo quality : a new light intensity measurement by a Lab-on-Chip technology was presented as a promising tool to evaluate embryo quality. As every year, the meeting combined a successful scientific programme with outstanding social events, encouraging exchanges between practitioners, students, scientists and sponsors. Therefore I would like to acknowledge the excellent organisation of this meeting by the Local Organising Committee: Prof. Dr. Heli Lindeberg (chair) and all her co-workers and students of the Department of Biosciences, University of Kuopio, Finland. European ET data collection and the biannual Newsletter. Both tasks represent important outcomes that can be delivered to our members and contribute to the AETE influence / vitality. Moreover, they really succeeded in managing the AETE society as President and Vice-president. Thanks a lot for your efficient as well as pleasant implication in the Board! Herewith, I would like to congratulate and welcome two new Board members who were elected during the general assembly: Hiemke Knijn (CRV, The Netherlands) and Dimitrios Rizos (INIA, Spain). They already started participating to the Board activities, as they proposed to overtake the previous mentioned tasks (ET data collection and Newsletter respectively). Finland. I also would like to thank all the sponsors and exhibitors for their financial support allowing us to organise a successful meeting like this. After 25 years of lifespan, our society experienced a new youthfulness of appearance with a new logo, voted by the board (see the Newsletter published in June). This outlines the implication of the Board, willing to modernize our society and to undertake new tasks concerning embryo transfer. As two major Board members (Sybrand Merton, President; Alfonso Gutierrez-Adan, Vice- President) left the Board after serving the AETE for 8 years, I would like to acknowledge their excellent active contribution to the Board : since many years, they were respectively responsible for the yearly Finally, I would like to acknowledge Rainer Saner, who accepted to become the new treasurer of our society and Frank Becker our new Vice President. Our next meeting (27 th ) will be held in Chester, UK on the 9th and 10th of September The Local Organising Committee, chaired by Dr Ian Kippax, has already started to organise this important event. Also most of the invited speakers already accepted to give a lecture. The scientific programme will be focussed on determinant steps for embryo development and maintenance of pregnancy: oocyte maturation, embryonic genome - 3 -

4 activation as well as the role of the endometrium. Therefore I am convinced that we can expect an interesting scientific meeting, including new perspectives in the field of embryo related biotechnologies. I wish you a happy new year and I am looking forward seeing you all in Chester next year. Claire ponsart President A.E.T.E January 2011 AETE BOARD MEMBERS Claire Ponsart, France, President claire.ponsart@unceia.fr Frank Becker, Germany, Vice President becker@fbn-dummerstorf.de Urban Besenfelder, Austria, Secretary urban.besenfelder@boku.ac.at Rainer Saner, Switzerland, Treasurer rsa@swissgenetics.ch Peter Vos, The Netherlands p.l.a.m.vos@uu.nl Ian Kippax, U.K. i.kippax@btopenworld.com Serge Lacaze, France Serge.lacaze@midatest.fr Jędrzej Jaskowski, Poland jasko@au.poznan.pl Hiemke Knijn, The Netherlands Hiemke.Knijn@crv4all.com Dimitrios Rizos, Spain drizos@inia.es A.E.T.E. Secretary Urban Besenfelder Reproduction Centre Wieselburg University of Veterinary Medicine Veterinaerplatz 1, A-1210 Vienna, Austria Tel: Fax : urban.besenfelder@boku.ac.at Upcoming Events American Embryo Transfer Association (AETA) & Canadian Embryo Transfer Association (CETA/ACTE) Joint Scientific Convention August 25-27, 2011 San Antonio, Texas, USA For information, please visit the CETA/ACTE web site at: or the AETA web site at: Continuing Opportunities Education The IETS foundation has an Educational Support Grant Program with the following purpose: To stimulate development of educational materials to meet the educational mission of the IETS Foundation. To encourage use of electronic technologies to deliver educational information regarding the development, application and research in embryo related technologies, including but not limited to, nuclear transfer, genetic enhancement of animals, embryo and gamete biology, and food safety and risk assessment of products arising from embryo biotechnology. All applications are due by the first of July. For guidelines and application form see: It would be very nice if we could get many applications also from Europe. Rainer Saner, Trustee of the IETS foundation (rsa@swissgenetics.ch) website:

5 A visual update of the last A.E.T.E. Scientific Meeting Dear Colleagues, I am taking this opportunity by putting some photos together (taken by a professional: Dr. Urban Besenfelder) to remind you the success (scientifically and socially) of the previous meeting of the Association that was held in Kuopio (Finland) at 10 th and 11 th of September It was a pleasure to visit Kuopio, a wonderful place in the middle of Finland. I would like to thank the Local Organising Committee, especially Dr. Heli Lindeberg for the organization of the fantastic meeting. I am confident that it will be another productive year for the Society and its members. The president and the board members of the society wishing you a happy new year Dimitrios Rizos Speakers at the Conference Participants of Workshop II Kuopio, Finland-September 2010 Gala Dinner at Peraniemen Kasino A welcome reception at Kuopio City Hall Typical Finnish Party at Jatkankamppa 5

6 Winner of the STUDENT COMPETITION spermatozoa per fertilized oocyte). We chose to evaluate the effect of protease inhibition after 2 as well as 6 h of incubation to Josine Beek, Inhibition of Serine Proteases by the Specific Inhibitor AEBSF has a Negative Influence on Sperm Motility and In Vitro Fertilization in Pigs Beek J., Maes D., Nauwynck H., Van Soom A. Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium. Proteases play a diverse role during the fertilization process as demonstrated by numerous studies in human and mice. In the pig, the sperm specific serine protease acrosin and its inhibitor are well documented. However, it still needs to be elucidated whether or not other serine, cysteine, metallo- or aspartic proteases are involved during porcine fertilization. Therefore, we investigated the effects of class-specific protease inhibitors on fertilization parameters using in vitro fertilization (IVF) in pigs. This abstract discusses the outcome of the inhibition experiments with the specific inhibitor of serine proteases 4-(2- aminoethyl) benzene sulfonyl fluoride hydrochloride (AEBSF). AEBSF is an irreversible inhibitor which is presumed to be active intracellular and has been used in cell culture in concentrations up to 250 μm. Ovaries of prepubertal gilts were collected at a local slaughterhouse. After in vitro maturation during 44 h, cumulus-oocyte-complexes were transferred to droplets of fertilization medium with 0, 100 or 250 μm AEBSF and inseminated with frozenthawed epididymal semen giving a final ratio of 600 spermatozoa per oocyte. After a co-incubation period of 2 or 6 h, presumed zygotes were vortexed to remove the protease inhibitor, washed four times in culture medium and cultured until 22 h post insemination. Subsequently, presumed zygotes (total n = 850) were fixed overnight with 4% paraformaldehyde and stained with bis-benzimide (Hoechst 33342, 10 μg/ml) to assess fertilization and polyspermy rate and to calculate the sperm penetration index (mean number of penetrated take account of a temporary inhibition or delay in sperm penetration which might be restored after 6 h of co-incubation. To evaluate the effect of AEBSF on sperm-zona binding, in vitro matured cumulusoocyte-complexes were denuded by vortexing, randomly assigned to three groups and fertilized in medium with 0, 100 or 250 μm AEBSF, respectively. Oocytes were inseminated giving a final ratio of 250 spermatozoa per oocyte to allow unambiguous counting of the number of spermatozoa bound to the zona pellucida. After 4 h of co-incubation, oocytes were washed to remove loosely bound spermatozoa, fixed overnight and stained with Hoechst (total n = 90). Sperm motility was evaluated via computer assisted sperm analysis after 0, 2, 4 and 6 h incubation of fresh ejaculated spermatozoa in fertilization medium with 0 or 250 μm AEBSF. Gamete toxicity was assessed by using SYBR-PI for spermatozoa and Hoechst-PI staining for oocytes. Presence of AEBSF during 6 h of gamete coincubation resulted in a significant reduction of all fertilization parameters compared to the control group, as presented in Table 1. Membrane integrity was not influenced by AEBSF. However, oocytes did show disintegration of the metaphase plate after 6 h of incubation with 250 μm AEBSF. Sperm motility was remarkably impeded by the inhibitor (Fig. 1). 6

7 Table 1. The penetration rate is almost 50% reduced in the presence of 100 μm AEBSF AEBSF (μm) Sperm penetration index Normal fertilization (%) Polyspermy (%) Penetration (%) Inhibition of penetration (%) a 25.1 a ± a ± a ± b 33.4 a ± b ± b ± b 6.6 b ± c ± c ± a,b,c Values (mean ± SEM) with different superscripts within the same column are statistically different (P <0.05) Furthermore, the mean number of sperm that bound to the zona pellucida (ZP) decreased by increasing concentrations of AEBSF. We suggest that the decline in fertilizing capacity of the spermatozoa is mediated through an AEBSF-induced decrease in sperm motility in combination with inhibition of proteins responsible for zona binding, like acrosin. Acrosin, a trypsin-like acrosomal protease, has been linked with secondary binding to the ZP as well as with dispersion of acrosomal content and limited zona lysis [1]. Yet, proteases can fulfill multiple functions in different pathways and protease inhibitors may therefore inhibit not only proteases present on the sperm membranes or in the acrosome, but also proteases involved in intracellular signaling. It has been described that serine proteases are involved in the control of sperm motility in mammals [2]. However, the nature and the localization of this serine protease remains to be established. Sperm motility is in part regulated by phosphorylation of flagellar proteins [3]. This pathway starts with activation of camp by influx of calcium and bicarbonate. The camp signal is then transducted to protein kinase A, which is involved in the phosphorylation process. Our hypothesis is that AEBSF inhibits a serine protease downstream of protein kinase A. Further experiments are planned to elucidate the localization and the function of the serine protease that influences boar sperm motility. Figure 1: Sperm Motility [1] Honda et al. Human Reproduction Update, 2002, Vol. 8, No.5, pp [2] De Lamirande and Gagnon. The Journal of Cell Biology, 1986, 102, pp [3] Turner. Reproduction, Fertility and Development, 2006, 18, pp

8 WORKSHOP 1 EMBRYO CRYOPRESERVATION: STATUS AND FUTURE DIRECTIONS Organiser: Catherine Guyader-Joly This workshop explored the question: Embryo cryopreservation what is or are the best solution(s)? The moderator started with an introduction reminding the aims of embryo cryopreservation (Box 1). The issue of embryo freezing was outlined in a second part. Two major cryopreservation protocols are used: conventional slow cooling and vitrification. Regarding the physico-chemical processes, they both aim to protect the cells from chilling injuries, intracellular ice formation, dehydration and toxic effects. Highly efficient methods have been developed for in vivo-produced bovine embryos, using different cryopreservation protocols. The outcome is mainly influenced by the physiological properties of embryos directly related to their chilling sensitivity. Box 1. Embryo cryopreservation: what are the aims? Arrest cellular metabolism Maintain structural and genetic integrity of the cells Achieve acceptable survival rate post-thaw or warming (pregnancy and calving rates) Protocol/technique must be consistent and reliable Freezing in vitro produced embryos is more problematic. Indeed, the first attempts to cryopreserve in vitro-produced bovine embryos revealed that they are more adversely affected by chilling than in vivo ones. Some fundamental characteristics of embryos (IVP, biopsied embryos, stage and quality) induce special problems for their cryopreservation. The most common approach to deal with the lower success rates of cryopreservation is to set up new procedures. It has been suggested that vitrification could be a solution to cryopreserve in vitro-produced embryos. An alternative approach is to modify the cells themselves to make them more resistant to cryopreservation. Culture conditions have a major influence on post-thaw survival of IVP embryos. Indeed, serum induces deviations of gene expression patterns related to apoptosis, oxidative stress and communications through gap junctions. Other culture medium supplements and environmental conditions were shortly mentioned. During the workshop, 5 speakers: Serge Lacaze, Sybrand Merton, Kirsi Kananen, Knut Roschlau and Giovanna Lazzari presented their data. This report summarized the main statements of their oral communication. Serge Lacaze: Results of sexing and direct transfer of biopsied frozen-thawed bovine embryos have been presented. Data have been obtained under field conditions in a commercial program between January 2000 and June A total of 671 grade 1 two different frozen media: 116 embryos were frozen in 1.5M EG with 40 % fetal calf serum and 555 embryos in 1.5M EG with 0.1M sucrose. After 10 seconds in air, straws were thawed in a waterbath at 25 C and used for direct transfer. Higher pregnancy rates are obtained with heifer recipients (46.3 %) compared with cow recipients (31 %). The overall pregnancy rates are 47 % with EG plus sucrose and 42 % with EG plus serum when recipients are heifers, so that EG plus sucrose has been chosen as actual freezing medium before direct transfer of biopsied embryos. No significant effects of embryo stage, interval between flushing and freezing and, number of biopsied embryos per flush were observed. Sybrand Merton: The choice of cryopreservation method in relation to pregnancy rate is determined by type, stage and 8

9 quality of embryos. In vivo embryos are more robust and the choice of cryopreservation technique is less important in comparison to in vitro produced embryos. In general, 5 to 10 % lower pregnancy rates are obtained with grade 2 embryos compared to grade 1. Effect of stage differs between in vivo and in vitro embryos: freezability is highest at the morula stage for in vivo embryos and at the blastocyst stage for IVP ones. Vitrification improves in vitro survival rates of IVP morulae but no improvement was observed for in vivo embryos. Some problems remain to be solved before applying vitrification in cattle for widespread use: open carriers, small volume of CPA solution surrounding the embryo and operator skillness influence greatly the results. A direct transfer technique is preferable with biopsied embryo. The choice of the freezing machine for slow freezing can strongly affect the survival of IVP frozen/thawed embryos. A higher risk is observed with liquid nitrogen freezers than alcohol ones. Final goal for IVP cattle embryos freezing should be vitrification in a closed straw, compatible with standard 0.25 ml straws which can be transferred directly. However, with large numbers of embryos, slow freezing procedures are still more efficient. Kirsi Kananen: Cumulative stress from biopsy and cryopreservation affects survival and pregnancy rates: a decrease of 10 to 30 percentage units is observed for biopsied fresh embryos and 20 to 40 percentage units for intact frozen in comparison to in vivo embryos transferred as fresh. L-ascorbic acid is a potent cellular antioxidant, abundant in bovine follicular fluid. Grade 1 day 7 in vivo and in vitro embryos were biopsied in holding medium containing ascorbic acid and slowly frozen in 1.5M EG plus ascorbic acid. IVP embryos presented a reduced apoptotic index with 0.2 mm ascorbic acid but no effect either on post-thaw survival rate or on the average total cell numbers of developed embryos were observed. After direct transfer of in vivo embryos treated with 0.1mM ascorbic acid, pregnancy rate (45 %) was not statistically different than control embryos but calving rate was increased (31 % vs. 22 %). Knut Roschlau: The procedure of embryo freezing for direct transfer has been described. In vivo produced embryos are frozen in 1.5M EG whereas IVP embryos are equilibrated in 1.5M EG supplemented with 0.1M sucrose. Seeding is induced at -6 C and straws are 9 maintained at -6 C during 10 min before being slowly frozen at a rate of 0.5 C/min until minus 32 C. Pregnancy rates vary from 55 to 60% for flushed embryos and from 35 to 40% for IVP embryos with great variations between donor cows. A recording system was developed by the International Embryo Transfer Society for straw labeling to minimize the risk of identification errors when an embryo is thawed and transferred to a recipient. The list of corresponding labels was exposed. No more than 13 codes have to be written on the straw or plug, cane and goblet (Photo 1)! Moreover, embryos frozen for direct transfer should be frozen in yellow translucent straws with a yellow plug and stored in a yellow identified goblet. Recommendations for thawing before direct transfer are to hold the straw 5 seconds in air then to put it in a 30 C water bath. Photo 1. Identification of the straws according to IETS (K. Roschlau) Giovanna Lazzari: Methods and results of conventional embryo freezing have been presented: glycerol and ethylene glycol are used as cryoprotectant for in vivo and in vitro produced cattle embryos, and in vitro produced buffalo and horse embryos. The methods applied at Avantea are very similar for the three species with the exception of the horse as only glycerol is used as cryoprotectant. Pregnancy rates in cattle, following transfer of frozen embryos, range from 50 to 55 % for in vivo produced and to 40 to 45 % for in vitro produced embryos. Culture conditions and particularly serum affect the in vitro survival after thawing of IVP embryos (Table 1). The buffalo is a species where very few data on embryo freezing are available in the literature, mainly because of the poor responses to

10 Table 1. Survival to freezing and thawing of IVP cattle embryos (G. Lazzari) N. frozen N. survived (%) N. hatched (%) BOC (21.4) a 6 (10.7) a BRL (49.0) b 13 (26.5) b SOF-serum 45 5 (11.1) d 0 (0.0) d SOF-BSA (80.9) c 32 (68.1) c conventional superovulation and estrus synchronisation protocols. In this species the in vitro embryo production techniques, from abattoir ovaries or from Ovum Pick Up, allow to obtain about 1 embryo per donor. Pregnancy rates following transfer of frozen embryos average 25 to 30 %. In the horse the OPU-ICSI technique is well established at Avantea so that pregnancy rates averaging % can be consistently obtained with in vitro produced frozen embryos. In the final part of the workshop, some additional feedbacks were given from the audience. An interesting discussion was centered on biopsied embryos: importance of the accurate biopsing method, role of the zona pellucida during the process of freezing and advantage of biopsy technique as assisted hatching were discussed. The last topic concerned vitrification procedures with reports of higher in vitro survival of vitrified IVP embryos. However, no data was available on pregnancy rates to confirm an improvement of in vivo viability. AETE Board Activity: Updating AETE Website One of the board activities is upgrading the website of the AETE. Therefore, we encourage the members of the society to put forward proposals, new ideas and/or suggestions. Please feel free to sent new ideas and proposals to Peter Vos, address: p.l.a.m.vos@uu.nl Thank you on behalf of the website committee. Ian Kippax, Claire Ponsart and Peter Vos Many thanks again to the organization and to all the participants for their contributions to this workshop. Catherine Guyader-Joly Corresponding addresses: - Serge Lacaze, Midatest, France: serge.lacaze@midatest.fr - Knut Roschlau, Masterrind, Germany: k.roschlau@t-online.de - Sybrand Merton, CRV, Netherlands: Sybrand.Merton@crv4all.com - Kirsi Kananen, Ovumia, Finland: kirsi.kananen@ovumia.fi - Giovanna Lazzari, Aventea, Italy : giovannalazzari@avantea.it - Catherine Guyader-Joly, UNCEIA, France : catherine.joly@unceia.fr 10

11 embryo per OPU session number of OPU sessions embryo per ET session number of flushes European Statistical data of bovine embryo transfer activity 2009 Sybrand Merton In this report a summary is given of the embryo transfer statistics of 2009 as presented during the 26th annual AETE meeting in September 2010 in Kuopio, Finland. The presented data includes information from 24 countries and is based on activities reported for breeding and commercial embryo production. Activities in relation to research purposes are not included. The presented data include numbers on embryo production (MOET, OPU-IVP and Genetic recovery-culled cows ) and transfers (fresh and frozen) for bovine and other species (sheep, swine, goat and horse). These data have also been forwarded to the International Embryo Transfer Association (IETS Data Retrieval Committee) for collation on a world-wide scale. A B Fig. 1: In vivo embryo production in Europe; A: Number of flushes B: Number of embryos per flush year year Embryo production The total number of flushed donors was 16,856 (excl. UK) which was a small decrease in activity compared to the previous year (-3.8%). This resulted in a collection of 106,495 transferable embryos (incl. UK). The mean number of transferable embryos per flush was Data from 2009 and previous years are shown in Figure 1. The total number of OPU sessions was 3,422. A decrease of 8.8% compared to last year. This resulted in a production of 6,096 transferable embryos. The mean embryo production was 1.78 embryos per session. Data from 2009 and previous years are shown in Figure A B ,5 2 1,5 1 0, year Fig. 2: In vitro embryo production in Europe; A: Number of OPU sessions B: Number of embryos per session 2003 year

12 OPU; n countries transferred embryos In 2009, five countries in Europe applied OPU-IVP for breeding and commercial purposes (Figure 3). embryos was 57% and 59% for the in vivo and in vitro embryos, respectively Fig. 3: In vitro embryo production in Europe; number of countries in which OPU-IVP was applied in breeding and commercial programmes year Fig.4: Total number of embryos transferred in Europe year The total number of Genetic recovery-culled cows sessions in 2009 were 867. An increase of 70% compared to the previous year. This resulted in a production of 1,557 transferable embryos. The mean embryo production was 1.8 embryos per session. The majority of these sessions were again as in previous years performed in Italy (see Table 1). Table 1: Application of OPU-IVP and Genetic recovery (ovaries from culled cows ) in Europe in Countries OPU-IVP sessions Netherlands 1, Germany 1,506 Italy Croatia 68 France Czech republic 2 Genetic recoveryculled cow sessions Distribution of the number of flushes and in vivo derived embryos transferred among the top 12 European countries is shown in Table 2. Table 2: Application of MOET in Europe; Top 12 European countries ranked according to the number of flushes performed in Countries Flushes Embryos Transferred France 6,030 29,218 Netherlands 2,788 16,481 Germany 2,445 14,066 Italy 1,992 10,500 England 4,857 Spain 687 2,198 Belgium 624 3,150 Switzerland 438 2,718 Finland 436 4,130 Ireland 338 1,734 Denmark 325 2,133 Czech republic 255 1,719 In total 114,148 transferable embryos were produced in Europe in A decrease of 7.4% compared to Embryo transfers The number of embryos transferred amounts to 100,678 (Figure 4) which almost equals the number of the previous year. The proportion of IVP embryos was 5.8%. The proportion of frozen 12

13 embryo transfers An overview of the application of MOET among the European countries is shown in Figure Sheep Swine Goat Equine year Fig. 6: Number of embryos transferred in Europe; other species. Summary A decrease in in vitro embryo production activities (- 8.8%) A decrease in total number of transferable embryos produced (-7.4%) Equal number of embryos transferred ET activities in other species declining >20,000 10,000-20,000 2,500-10,000 <2,500 No activity (reported) Fig. 5: Application of MOET in Europe (number of embryos transferred). Acknowledgements: I would like to thank all the participants who collected the embryo transfer statistics for their country and helped me and the society for the past eight years. Since I resigned from the board last September, Hiemke Knijn will take over this task. I wish her all the best and I am already looking forward to the results of Sybrand Merton Other species Data for embryo transfer activities in sheep, swine, goat and equine are shown in Figure 6. The number of embryos transferred in sheep decreased dramatically and was more than halved compared to last year. Embryo transfer activities in equine stabilised, although only 5 countries reported activities in 2009 compared to 7 in As previously, Italy reported also the production and transfer of in vitro produced embryos (n=60). Activities in swine remained limited and no activity at all was reported for goat. 13

14 The 27th Scientific Meeting of the A.E.T.E city on the banks of the river Dee on the border with Wales. The city is a great staging point for exploring many attractions of the city and the surrounding country in England and Wales. Will be held in Chester United Kingdom 9 TH -10 TH SEPTEMBER 2011 Invitation On behalf of the European Embryo Transfer Association the local organizing committee cordially invites you to the 27th scientific meeting of the organization in Chester, United Kingdom, from the 9 th to the 10 th of September Chester The Local Organizing Committee will be chaired by - Ian Kippax, i.kippax@btopenworld.com The Conference Location This years conference will be based at the recently refurvished Queen Hotel which is close to the city centre, near the railway station. Chester has many attractions for visitors, the history for those who are interested, with many Roman sites and museums. For those who like shopping there is the famous Rows Shopping centre and close by are the cities of Liverpool and Manchester. Chester has its own racecourse, and on the weekend of the conference there is a meet at Chester race course on the Roo Dee. Welcome to the Chester-UK Welcome to the north west of England, to the county of Cheshire and to the town of Chester. Chester was built as a roman fortress. It is a walled 14

15 We hope to spend an evening at Peckforton Castle near by, and an evening at Chester Zoo. 1 REGISTRATION FEES How to travel Chester is easy to get to with 2 airports, Liverpool and Manchester nearby. There is also a motorway network and good road access for those who wish to come by car and then spend some time touring. Chester 2011 Full/Associate Member Before 15th July 2011 Full/Associate Member After 15th July 2011 Student Member Before 15th July 2011 Student Member After 15th July Membership Fee Members who pay their annual fee but do not attend the Meeting will receive a copy of the proceedings Euros This price includes: - membership fee - participation at the Meeting (two full days) - two workshops - published proceedings - lunch and coffee breaks - social events 1 Fees for Sponsoring AETE Meeting Come to Chester and take some time to see what the northwest of England has to offer, Chester particularly, but also Manchester and Liverpool or the National Parks, The Lake District, The Peak District or the Snowdon National Park. We look forward to seeing you in 2011 in Chester. Ian kippax Chairman of the Organizing Committee Language The official language of the conference is English. Main Sponsor General Sponsor Exhibitors Supporters Euros Euros Euros Euros Costs for advertisement in the Newsletter (2 issues) for one year (mailed to ~700 members) Full color back page Full inside color page Half inside color page 800 Euros 600 Euros 400 Euros Scientific Secretariat AETE board 15

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