48. Jahrestagung Physiologie und Pathologie der

Reprod Dom Anim 50 (Suppl. 1), 1–77 (2015); doi: 10.1111/rda.12498 ISSN 0936–6768

48. Jahrestagung Physiologie und Pathologie der Fortpflanzung und gleichzeitig 40. Veterinär-Humanmedizinische Gemeinschaftstagung Februartagung 2015 Zürich, Schweiz, 11. – 13. Februar 2015 48th Annual Conference Physiology and Pathology of Reproduction and simultaneously 40th Joint Congress of Veterinary and Human Medicine February Conference 2015 Zurich, Switzerland, 11th – 13th February 2015

ETH Zurich Animal Physiology, Institute of Agricultural Sciences, Department of Environmental Systems Science University of Zurich Clinic of Reproductive Medicine, Vetsuisse-Faculty Trägergesellschaften der Tagung: Deutsche Veterinärmedizinische Gesellschaft e.V. (DVG), Fachgruppe Fortpflanzung und ihre Störungen Deutsche Gesellschaft für Reproduktionsmedizin e.V. (DGRM)

Image source: http://www.math.ethz.ch/~ocurdin/bild1.jpg


Grusswort zur Februartagung 2015 Liebe Kolleginnen und Kollegen, Wir freuen uns sehr, Sie zur 48. Jahrestagung Physiologie und Pathologie der Fortpflanzung und gleichzeitig 40. Veterinär-Humanmedizinische Gemeinschaftstagung in Zürich begrüßen zu dürfen. Die Tierphysiologie der ETH Zürich und die Klinik für Reproduktionsmedizin der Universität Zürich richten gemeinsam die diesjährige Februartagung 2015 aus. Wir möchten gern dazu beitragen, die naturwissenschaftlich orientierte Grundlagenforschung und die klinisch orientierte Reproduktionsmedizin näher zusammenzurücken und die Zusammenarbeit zwischen beiden Gebieten weiter zu stärken. Ein besonderes Anliegen ist uns die aktive Teilnahme von jungen Nachwuchswissenschaftlerinnen und wissenschaftlern. In Anerkennung ihrer Tätigkeit haben wir ihnen möglichst viel Präsentationsmöglichkeiten eingeräumt. Als Mittelpunkt des wissenschaftlichen Austausches haben wir die Themen Metabolische Programmierung sowie Einflüsse der Inflammation auf die Fertilität gewählt. Wir konnten als Hauptreferenten mit Claire Wathes, Anne Navarrete-Santos, Klaus Steger und Mats Troedsson herausragende Fachleute gewinnen, die human- und veterinärmedizinisch herausfordernde wissenschaftliche Aspekte bei verschiedenen Spezies präsentieren werden. Die Februartagung ist seit Jahrzehnten die wichtigste Tagung zum Thema Physiologie und Pathologie der männlichen und weiblichen Fortpflanzung im deutschsprachigen Raum. Ein Charakterzug des Gastgeberlandes Schweiz ist sein eleganter Umgang mit vier verschiedenen Landessprachen (deutsch, französisch, italienisch, rätoromanisch) neben der Wissenschaftssprache englisch. Im Wissen um die sprachliche Vielseitigkeit der Teilnehmerinnen und Teilnehmer bitten wir in diesem Jahr um englischsprachige Vorträge. Dies soll nicht von deutsch- und anderssprachigen Diskussionen abhalten, sondern allen erlauben, Einsicht in die neuesten Forschungsergebnisse zu erhalten. Der Erfolg der Tagung hängt ganz maßgeblich von den Teilnehmerinnen und Teilnehmer ab, die durch Ihr Interesse und Engagement am wissenschaftlichen Diskurs teilnehmen. Wir freuen uns daher auf rege Diskussionen mit Ihnen, heißen Sie ganz herzlich in Zürich willkommen und wünschen Ihnen eine vielseitig bereichernde Tagung. Mit freundlichen Grüßen,

Prof. Dr. Susanne E. Ulbrich

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Prof. Dr. Heinrich Bollwein


Welcome to the February Conference 2015 Dear collegues, th

It is our great pleasure to welcome you in Zurich to the 48 Annual Conference Physiology and Pathology th of Reproduction and simultaneously 40 Joint Congress of Veterinary and Human Medicine. The Animal Physiology of ETH Zurich and the Clinic of Reproductive Medicine of the University of Zurich are jointly hosting the February Conference 2015. We specifically aim at closing ranks between basic research in natural sciences and the clinically oriented reproductive medicine as well as to further strengthen the collaboration between the two areas. The active participation of young scientists is of particular concern to us. In recognition of their excellent work, we have granted them as much presentation options as possible. We have chosen the topics of metabolic programming and effects of inflammation on fertility as a focus of our scientific exchange. With Claire Wathes, Anne Navarrete-Santos, Klaus Steger and Mats Troedsson, we were able to win outstanding experts as keynote speakers. They will present scientifically challenging aspects of human and veterinary medicine in different species. For decades, the February conference has been the most important german-speaking conference on physiology and pathology of the male and female reproduction. One important characteristic of this year´s host country Switzerland is the elegant handling of four different national languages (German, French, Italian, Rhaeto-Romanic) in addition to English as scientific language. Knowing the linguistic diversity of the participants, we decided to ask the speakers for English lectures. This is not meant to discourage discussions in German and other languages. It simply allows everyone to gain insight into the latest research. The success of the conference also crucially depends on its auditorium. You are therefore kindly invited to actively participate in the scientific discussions with the speakers and the collegue researchers. We friendly welcome you in Zurich, wish you a nice stay and hope you enjoy a successful conference.

Kind regards,

Prof. Dr. Susanne E. Ulbrich

Prof. Dr. Heinrich Bollwein

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Table of Contents Congress Sponsors ............................................................................................................... 5 Key Lectures .......................................................................................................................... 6 Short Presentations ............................................................................................................... 9 Session 1: Endocrinology & Reproduction ....................................................................................... 9 DGRM prize session ...................................................................................................................... 11 Session 2: Metabolic programming & Female reproduction .......................................................... 13 PhD Student competition ............................................................................................................... 15 Förderverein Biotechnologieforschung e. V. (FBF) (Association for Biotechnology Research) Symposium .................................................................................................................................... 16 Session 3: Inflammation & Fertility ................................................................................................. 18 Postdoc competition ....................................................................................................................... 21 Session 4: Gametes & Embryos .................................................................................................... 22

Poster .................................................................................................................................. 26 Biotechnologies in reproduction ..................................................................................................... 26 Cycle, gravidity and peripartum period .......................................................................................... 32 Embryonic and foetal development ................................................................................................ 47 Gametes......................................................................................................................................... 50 Genetics, genomics, biochemistry ................................................................................................. 53 Inflammation and fertility ................................................................................................................ 60 Other topics .................................................................................................................................... 68

Index of authors ................................................................................................................... 73

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Congress Sponsors

Biokema SA

Crissier, Switzerland

GOLD

Labotect Labor-Technik-Göttingen GmbH

Göttingen, Germany

GOLD

MSD Animal Health GmbH

Lucerne, Switzerland

GOLD

Provet AG

Lyssach, Switzerland

GOLD

SUISAG

Sempach, Switzerland

GOLD

VIRBAC Schweiz AG - Animal Health

Glattbrugg, Switzerland

Virbac Tierarzneimittel GmbH

Bad Oldesloe, Germany

Zoetis Schweiz GmbH

Zürich, Switzerland

GOLD

Gynemed GmbH & CO.KG

Lensahn, Germany

SILVER

IMV Technologies

Munich, Germany

SILVER

Swissgenetics

Zollikofen, Switzerland

SILVER

Anklin AG

Binningen, Switzerland

BRONZE

Eickemeyer Medizintechnik für Tierärzte AG

Appenzell, Switzerland

BRONZE

Minitüb GmbH

Tiefenbach, Germany

BRONZE

Olympus Schweiz AG

Volketswil, Switzerland

BRONZE

PHYSIA GmbH

Neu-Isenburg, Germany

BRONZE

scil animal care company GmbH

Viernheim, Germany

BRONZE

GOLD

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Key Lectures K1 The impact of metabolic stress on immunity and fertility in dairy cows DC Wathes, T Swangchan-Uthai, CF Oguejiofor and Z Cheng Department of Production and Population Health, Royal Veterinary College, North Mymms, Hatfield, UK The periparturient period is a challenging time for the immune system of a typical dairy cow. Maternal insulin resistance increases in late pregnancy to promote transfer of nutrients initially across the placenta and subsequently across the mammary gland into milk. The cow is generally unable to meet these increasing metabolic demands through feed intake and so mobilizes body tissue. This is associated with alterations in blood metabolites and metabolic hormones: NEFAs and BHB concentrations rise while glucose, insulin and IGF1 concentrations fall. All of these changes are associated with a reduced capacity of the immune system to respond to infection: circulating neutrophil concentrations decline and their individual capacity to respond to a challenge is also diminished. At the same time the process of giving birth opens the cervix to the external environment and the reproductive tract is exposed to an influx of bacteria. Placental separation leaves the caruncular regions without their normal protective epithelial layer and a degree of endometrial infection is almost inevitable. Clinical and subclinical endometritis develop in 20-30% of all postpartum dairy cows and are associated with delays in uterine involution, disrupted oestrous cyclicity and a reduced likelihood of conception. The first key question is how quickly and effectively can the cow mount an immune response against the invading bacteria? This involves activation of the toll-like receptors to initiate an inflammatory response typically characterised by the production of pro-inflammatory cytokines, type I interferons, chemokines and antimicrobial proteins and recruitment of leucocytes, monocytes and lymphocytes into the inflamed endometrium. The second question is how soon after will the inflammation be resolved, returning the endometrium to a state where it is again able to support a developing conceptus? Both of these issues are influenced by a combination of systemic and local factors. When the postpartum cow is in a poor energy balance status, organs throughout the body are affected. At a local level there is evidence for alterations in the IGF family of growth factors which may be associated with delays in uterine repair. The high concentrations of lipid present in the endometrium may reduce macrophage and neutrophil functionality. During infection the balance of lipid mediators produced, including prostaglandins, and their release patterns are altered. This potentially contributes to extended luteal phases. A final consideration is whether the cow is exposed to any other sources of infection during this

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period. There is evidence that some viruses can cause immunosuppression. For example, we have recently shown that endometrial infection with ncpBVDV inhibits the expression of many innate immune genes that typically increase in response to both bacterial lipopolysaccharide (LPS) and the implantation signal interferon tau. This includes genes involved in pathogen recognition, inflammatory response, interferon response, chemokine activity, tissue remodelling, cell migration and cell death/survival. These changes are likely to increase the risk of infection and also interfere with the pregnancy recognition signal. We are grateful to the Wellcome Trust, the Royal Thai Government and the Commonwealth Scholarship Commission for support. K2 Mechanisms of metabolic and hormonal adaptation in preimplantation embryos - or how the embryo survives a diabetic pregnancy A Navarrete Santos, J Gürke, K Grybel, J Knelangen, M Pendzialek, M Schindler, R Thieme and B Fischer Department of Anatomy and Cell Biology, Martin Luther University Halle-Wittenberg Faculty of Medicine, Halle (Saale), Germany Embryogenesis of the rabbit is a particularly suitable model for the human. It allows molecular studies on preimplantation embryos and the influence of maternal diseases on embryo development. Like in women, a diabetes mellitus type 1 during the peri-conceptional period leads to subfertility with metabolic and hormonal changes in reproductive organs of female rabbits. As a consequence, the preimplantation rabbit embryo adapts to the diabetic uterine environment by distinct changes in metabolism. In diabetic rabbits an oversupply of maternal nutrients such as glucose, amino acids and lipids in uterine secretions, correlates with a non-physiological high accumulation of lipids and enforced protein synthesis in the embryo. However the embryo does not “profit” from this overfeeding indicated by a delayed development and an increase in apoptosis. The loss of maternal insulin is highly critical for gastrulation as maternal insulin and insulin-like growth factors (IGF) promote the development, growth and differentiation of the embryo. IGFs regulate differentiation-specific gene expression and balance embryonic energy and glucose metabolism. The conversion of embryonic metabolism from insulin- to adiponectin-dependent glucose metabolism allows the embryo to adapt to a diabetic environment. Paracrine adiponectin takes over insulin functions in glucose uptake, but also stimulates PPAR expression in an excess. Furthermore, embryos from diabetic rabbits show a higher ROS production and various protein modifications, indicating that embryos are metabolically imbalanced and exposed to cellular stress. We conclude that the mammalian preimplantation


embryo is sensitive to maternal metabolic disorders but can adapt and survive due to an endogenous hormonal and molecular network. This work was supported by the German Research Council (DFG; grant number NA 418/4-2), the EU (FP-7: Epihealth-278418, Epihealth NET-317146, COST 1101, 1201) and the Wilhelm Roux Programme of the MLU Faculty of Medicine. K3 Breeding induced endometritis: Physiology or Pathology? MHT Troedsson Maxwell H. Gluck Equine Research Center University of Kentucky, Lexington, KY, USA The equine endometrium reacts to breeding with a transient innate inflammation, characterized by the expression of pro- and anti-inflammatory cytokines, activation of the complement cascade, and an influx of polymorphonuclear neutrophils (PMNs) into the uterine lumen. The onset of inflammation starts already within 0.5-1 hours after deposition of semen into the uterine lumen, and is resolved within 36 to 48 hours in most reproductively sound mares. In vitro studies demonstrating neutrophil extracellular traps (NETs), PMN phagocytosis of spermatozoa, and an apparent regulation of this process mediated by specific seminal plasma proteins, suggests that breeding-induced endometritis plays an important role in selective sperm transport to the oviduct and elimination of dead sperm from the uterus. Recent studies showed an increase in mRNA expression of modulating cytokines (IL-6, IL-10 and IL-1RN) within 6 hours after insemination, suggesting a role of these cytokines in the transient nature of the inflammation. Altogether, these observations suggest that breeding-induced endometritis is a fine tuned transient inflammation, assisting the uterus in securing sperm transport of viable spermatozoa to the oviduct and ensuring a rapid clearance of excess spermatozoa and inflammatory products from the uterus well before the embryo is transported into the uterine lumen at 96 hours after fertilization. In a subpopulation of mares, the breeding-induced inflammation fails to resolve before the embryo enters the uterus. Pregnancy rates in these mares are lower than in mares that resolve the inflammation in a timely fashion, suggesting that the persistent inflammation has a negative effect on normal development and viability of the conceptus. The exact mechanism involved in the development of a persistent inflammation is not fully understood. Impaired myoelectrical activity has been implicated in delayed uterine clearance, and an up-regulation of iNOS with subsequent accumulation of NO in the uterus was suggested to be involved in this development. Recent studies suggest that an imbalance of pro- and antiinflammatory cytokines between 2 and 6 hours after breeding may contribute to the development of persistent inflammation. Messenger RNA expression of proinflammatory cytokines was similar in all mares in

response to breeding, but mares with a history of persistent breeding-induced endometritis had significantly lower mRNA expression of the modulating cytokines IL-6, IL-10, and IL-1RN at 6 hours after breeding, compared to mares that resolved the inflammation in a timely fashion. In conclusion, breeding-induced endometritis is a physiological response to breeding, involving a fine tuned inflammatory mechanism that ensures resolution within 36 to 48 hours. Mares with sub-normal expression of modulating cytokines at 6 hours after breeding and an over expression of iNOS, develop delayed uterine clearance which is associated with subfertility. K4 Sperm epigenome: paternal contribution to the embryo K Steger Biomedizinisches Forschungszentrum der Justus-LiebigUniversität Gießen, Germany Sektion Molekulare Andrologie der Klinik für Urologie, Kinderurologie und Andrologie Epigenetic mechanisms regulate which genes will be expressed at a specific time point in a distinct cell-type. Mother´s lifestyle is well-known to influence the distribution pattern of epigenetic marks that may be transmitted to her offspings (1). Recently, it has been reported that father´s lifestyle may also affect gene expression in the offsprings (reviewed in 2) suggesting that sperm must transmit epigenetic information to the oocyte at fertilization. Epigenetic processes influence gene expression via DNA-methylation, histone modifications and short non-coding RNAs (reviewed in 3). DNA-methylation occurs at cytosine residues within CpG dinucleotides (CpG-islands). Apart from imprinted genes exhibiting a parent-of-origin-dependent gene expression, most gene promoters are hypomethylated resulting in transcriptional activation, while promoter hypermethylation is associated with gene silencing. The most extensively studied histone modifications include acetylation of lysines, mono-, di- or trimethylation of lysines and mono- or dimethylation of arginines. While histone acetylation is generally associated with chromatin expansion and gene transcription, the effect of histone methylation on gene expression is either activating or repressing depending on the position of the amino acid and the number of methylated residues. Interestingly, sperm cells exhibit an additional epigenetic mechanism, the replacement of histones by protamines, which results in gene silencing of the entire genome. As histone-toprotamine exchange is incomplete, remaining histones carrying epigenetic marks have been hypothesized to be involved in the regulation of gene expression in the early embryo. Consequently, several studies performing deepsequencing have been published within the last six years (reviewed in 4). Indeed, spermatozoal nucleosomes have been reported to be enriched at loci of developmental importance, namely imprinted gene clusters, microRNA

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clusters, HOX gene clusters and promoters of developmental transcription factors (5,6). Subsequently, retained nucleosomes have been attributed to GC-rich sequences lacking DNA-methylation (7,8). Recently, nucleosome retention in sperm was reported to be associated with distal intergenic regions and introns, centromere repeats and retrotransposons, whereas 5´ and 3´ untranslated regions as well as transcriptional start and termination sites have been demonstrated to reveal nucleosome depletion (9,10). Interestingly, the preparation method seems to play an important role on the outcome of deep sequencing studies (10).

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References: (1) Wolff et al. 1998 FASEB J 12:949-957, (2) Hughes 2014 Nature 507:22-24, (3) Schagdarsurengin et al. 2013 Nature Reviews Urology 9:609-619, (4) Saitou & Kurimoto 2014 Dev Cell 30:6-8, (5) Hammoud et al. 2009 Nature 460:473-478, (6) Arpanahi et al. 2009 Genome Res 19:1338-1349, (7) Vavouri & Lehner 2011 PLoS Genetic 7e1002036, (8) Erkek et al. 2013 Nat Struct Mol Biol 20:868-875, (9) Samans et al. 2014 Dev Cell 30:23-35, (10) Carone et al. 2014 Dev Cell 30:11-22.


Short Presentations Session 1: Endocrinology & Reproduction SP1 Genomic evaluation of female fertility in Brown Swiss dairy cattle Genomische Analyse der weiblichen Fruchtbarkeit beim Braunvieh C Baes1,2, B Bapst2, C Flury1, H Signer-Hasler1, D Garrick3, C Stricker4, F Schmitz-Hsu5, and B Gredler2 1 Bern University of Applied Sciences, Zollikofen, Switzerland; 2Qualitas AG, Zug Switzerland; 3Iowa State University, Ames, IA, USA; 4agn Genetics, Davos, Switzerland; 5Swissgenetics, Zollikofen, Switzerland Genetic improvement of fertility traits is challenging due to fuzzy phenotypes, the polygenic nature of the trait, antagonistic genetic correlations to milk production traits and notoriously low heritability. Nevertheless, many countries have introduced genetic evaluations for fertility traits. Gredler and Schnyder implemented a new model for reproductive performance for Swiss Brown Swiss cattle in 2013, which includes non-return-rate (heifers and cows), days to first service, and interval between first and last insemination (heifers and cows). Medium (50K; 54,609 SNP) and high-density (HD; 777,962 SNP) arrays can be applied in genome-wide association studies (GWAS) to identify causative mutations responsible for economically important traits, including fertility. Whole genome sequence data is expected to provide even more concise information for GWAS, because causal variants are included in the data per se. Using whole genome sequence information of 100 Brown Swiss bulls, the autosomal sequence of 12,000 genotyped Brown Swiss animals was imputed. The accuracy of imputation from 50K and HD arrays to sequence genotypes was evaluated by calculating concordance between true and imputed genotypes. A GWAS was conducted with special emphasis on previously identified genome regions shown to be associated with fertility. We provide evidence that specific regions of the genome could be associated with fertility traits. These regions may be candidates for further fine mapping studies. The ASR and the CTI funded this project. Intergenomics partners provided array genotypes; the 1000 Bull Genomes Project and Gene2Farm provided sequence data. SP2 Effects of treatments with GnRH and/or PGF2α during early puerperium on fertility in dairy cows Auswirkungen der Behandlung mit GnRH und/ oder PGF2α im Frühpuerperium auf die Fruchtbarkeit von Milchkühen J Krohn1, S Haß1, P Georgiev2, A Wehrend1

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Klinikum Veterinärmedizin, Klinik für Geburtshilfe, Gynäkologie und Andrologie der Groß- und Kleintiere, Justus-Liebig-Universität Gießen, Germany; 2Faculty of Veterinary Medicine, Thrakia University, Stara Zagora, Bulgaria Hormone treatments in the puerperal period are very popular but the effects documented in literature are controversial. In this study the effect of administration of GnRH and/ or PGF2α up to 21 days post partum in cows with (dp=disturbed puerperium) and without (pp=physiological puerperium) a metritis was investigated focusing on further puerperal development and fertility. 305 cows were included in the study. The PGF2α group (pp1: n=65, dp1: n=41) was treated with 25 mg Dinoprost between 11-14 days post partum. The GnRH group (pp2: n= 47, dp2: n=50) received 50 µg Gonadorelin and seven days later 25 mg Dinoprost. In the control group (pp3: n=48, dp3: n= 54) 1 ml 0.9% saline was administered between 11-14 days post partum. All injections were given intramuscularly. Gynaeco-logical examinations were carried out at the day of first treatment and between 25-31 days post partum and according to clinical sings. Incidence of ovarian cysts and development of an endometritis after 60 days of lactation was recorded. The body condition (BC) was measured by scanning the back fat thickness about two weeks before parturition, 14-16 and 60-66 days after parturition. Milk production was estimated by calculating the mean daily production. The following fertility parameters were evaluated: number of inseminations, first service conception rate, pregnancy rate, interval to first service (IFS), insemination index and calving to conception interval. Neither Dinoprost nor GnRH-treatment had a significant influence on uterine involution (p > 0.05) or reduction of pathological vaginal discharge. No influence of the hormone treatment on the number of inseminations, first service conception rate, calving to conception interval, pregnancy rate or incidence of ovarian cysts was detected in the cows with disturbed and physiological puerperium. The effect of hormone treatments on puerperal development and fertility did not depend (p > 0.05) on milk production, BC or its decrease during early lactation. The results indicate that treatments with PGF2α or a combination of GnRH and PGF2α during the early puerperal period do not affect uterine involution and fertility in dairy cows no matter if the cows showed a metritis or not. SP3 Establishment of pregnancy is associated with TSHlevels on the day of ovulation Trächtigkeitsetablierung ist assoziiert mit den TSHWerten am Tag der Ovulation M Meyerholz1, K Mense1, M Lietzau1, A Kassens2,

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M Linden3, M Raliou4, C Richard4, O Sandra4, M Hoedemaker1, M Piechotta1 1 Clinic for Cattle and 2Reproduction Unit, University of Veterinary Medicine Hannover, Germany; 3Leibniz University, Faculty of Mathematics and Physics, Hannover, Germany; 4INRA, UMR1198, Biologie du Développement et Reproduction, Jouy-en-Josas, France Thyroid-stimulating hormone (TSH) is the main regulator of thyroid gland function, and elevated TSH in humans is known to be associated with infertility. The aim was to establish an ELISA for measurement of bovine TSH and to compare TSH serum concentrations of early pregnant (n=19) and non-pregnant (n=19) Holstein Friesian heifers. For the TSH ELISA, an antibody targeted against bovine TSH (anti-bovine TSH, 1:10 pre-diluted, AFP642482Rb, obtained from the National Hormone & Peptide Program (NHPP), NIDDK and Dr. Parlow) was diluted (1:2500). The applied standard curve ranged from 0.2–100 ng/ml bovine TSH (AFP-8755B, obtained from the NHPP), and biotin-labeled TSH was used to detect binding with subsequent addition of streptavidin horseradish peroxidase solution. The intraassay CV was determined by measuring one bovine sample 20 times. Serum samples of 19 matched pairs were used to compare pregnant and non-pregnant heifers. Blood was taken from the jugular vein on the Day of ovulation (defined by ultrasound; Day 0) and Days 14 and 18. Pregnancy status was defined as follows: pregnant = progesterone [ng/ml] >2.0 on Day 18 and successful conceptus collection or detectable embryonic heartbeat in ultrasound investigations on Day 42; non-pregnant = progesterone 0.05) during the study period in the control group. These observations suggest that endotoxins induce a transient decrease in uterine blood supply without having any effects on viability of the embryo. P97 Effects of progesterone, estradiol 17-ß and prostaglandin F2α on myometrial contractility of inflamed uteri in cows immediately after parturition Effekte von Progesteron, Östradiol und Prostaglandin F2α auf die myometriale Kontraktilität entzündeter Uteri von Kühen unmittelbar nach der Geburt L Górriz Martín1, H Bollwein2, M Piechotta1, L Holzhausen1, C Pfarrer3, G Hirsbrunner4, C Keller5, M Hoedemaker1, M Heppelmann1 1 Clinic for cattle, University of Veterinary Medicine, Hannover, Germany; 2Clinic of Reproductive Medicine, Vetsuisse Faculty, University of Zurich, Switzerland; 3 Department of Anatomy, University of Veterinary Medicine, Hannover, Germany; 4Clinic for Ruminants, University of Berne, Berne, Switzerland; 5Cantonal Veterinary Service of Berne, Berne, Switzerland Myometrial contractility is crucial for uterine involution after parturition in cows and seems to be affected by sexual hormones such as progesterone (P4), estrogens and prostaglandins. Since inflammations induced by endotoxins released by gram-negative bacteria are common after calving, it was the aim of this study to examine the effects of concentrations of P4, estradiol 17-ß (E2ß) and 15-keto-13,14-dihydro-PGF2α (PGFM) in blood plasma on in vitro contractility of the myometrium in inflamed uteri immediately after parturition. After placental expulsion (within 12 h after calving), 5 µg/mL LPS dissolved in 1 L 0.9% NaCl isotonic solution at 37°C was infused transcervically into the uterus of 12 cows. Three hours after infusion, plasma blood samples were taken and the cows were euthanized to collect myometrial stripes from the longitudinal [LL] and circular [CL] layers. Spontaneous contractility (SC) and contractility induced (IC) by increasing concentrations of oxytocin (Oxy), prostaglandin F2α (PGF), ionized Ca2+ (Ca) or nothing (Con) were recorded in an organ bath during 9 intervals of 30 minutes each (5 and 4 intervals for SC and IC, respectively). Mean amplitude (MA) and area under the curve (AUC) were calculated for each interval to quantify uterine contractility. Blood P4, E2ß and PGFM values averaged 0.4 ± 0.1 ng/mL, 87.6 ± 39.5 pg/mL, and 2154.1 ± 886.0 pg/mL, respectively. There were no differences (P > 0.05) in contractility between LL and CL. Stimulation with Oxy caused higher contractile AUC and MA values (P ≤ 0.05) than PGF, Ca and Con (Oxy>PGF>Ca, Con). Only during the last 30


min of spontaneous contractility recordings MA and AUC correlated with PGFM (r = -0.60; P ≤ 0.05). Consequently, myometrial contractility of inflamed postpartum uteri was not affected by P4 or E2ß in blood plasma. Surprisingly, cows with high plasma levels of PGFM showed lower myometrial contractility after 2 h of incubation in the organ bath. P98 Is a single exfoliative cytology representing the entire endometrium in cattle? Sind exfoliativ-zytologische Befunde der Gebärmutter beim Rind abhängig vom Entnahmeort? J Egberts1,2, J Detterer2, A Park2, S Meinecke-Tillmann1 1 Institut für Reproduktionsbiologie, Tierärztliche Hochschule Hannover, Germany; 2Besamungs- und ETStation Georgsheil, VOSt Südbrookmerland, Germany Uterine cytology is considered to be a reliable way to diagnose subclinical endometritis in cattle, although the question has been raised of whether one sample is reflecting the status of the entire uterus. Therefore, the aim of the study was to determine whether the results of exfoliative cytology of a single intrauterine localization are representative for the whole organ. Using the cytobrush method 20 German Holstein cows (1st-8th lactation) were sampled at three intrauterine localizations during interestrus (day 7-12; pre-examination of embryo transfer donors). These included both uterine horns about 1.5 cm cranial to the bifurcation, resp., as well as the uterine body about 0.5 cm cranial to the internal os of the cervix. The obtained cellular material was spread out on microscope slides, and after staining with DiffQuick® the cytological status was assessed. The percentage of polymorphonuclear neutrophils (PMN) was determined per 300 cells per slide. Although the mean percentages of PMN found in the uterine body (4.8±11.4), right uterine horn (4.6±8.5) and left uterine horn (4.8±9.2) of the animals did not differ significantly (P=0.75), intraindividual differences became obvious. Subclinical endometritis (SE) was diagnosed in 30 % of the cows. But only in 3 of these 6 animals the results of exfoliative cytology of all three sampling locations within the uterus led to the diagnosis of SE. In the remaining 3 animals the PMN threshold was only reached in one or two of the three sampling locations (two uterine horns, single uterine horn and corpus uteri or only one uterine horn, resp.). Therefore it might be advisable to take samples from more than one intrauterine localization in order to verify a subclinical endometritis. P99 Rearing intensity during early postnatal life has longterm effects on pancreatic ß-cells in Holstein bull calves S Wiedemann1, L Prokop1, R Lucius2, M Maccari3, H-J Kunz4, M Kaske5 1 Department of Animal Health and 2Institute of Anatomy, University of Kiel, Germany; 3University of Veterinary

Medicine Hannover, Germany; 4Chamber of Agriculture of Schleswig-Holstein, Germany; 5Department for Farm Animals, University of Zurich, Switzerland A permanent effect of early postnatal development on growth and function of tissues and organs has been reported for various mammalian species. The objective of this study was to assess the impact of two different rearing strategies during the first 3 weeks (wk) of life on insulin producing pancreatic beta cells in male Holstein calves. During the first 3 wk of life, calves were fed either intensively (INT; ad libitum milk feeding; individual hutches; N=21) or according to an established restrictive rearing protocol (CON; 4 L milk/d during wk 1 in hutches, 720 g/d milk replacer from d 8–21 in group pens; N=21). Thereby, the average daily gain was higher in INT-calves compared to CON calves during the first 3 wk of life (1.28 vs. 0.38 kg/d; p