Association between days post conception and lactation persistency in dairy cattle.

Determining the optimal insemination moment for individual cows is complex, particularly when considering the impact of pregnancy on milk production. The effect of pregnancy on the absolute milk yield has already been reported in several studies. Currently, there is limited quantitative knowledge about the association between days post conception (DPC) and lactation persistency, based on a lactation curve model, and, specifically, how persistency changes during pregnancy and relates to the days in milk at conception (DIMc). Understanding this association might provide valuable insights to determine the optimal insemination moment. This study, therefore, aimed to investigate the association between DPC and lactation persistency, with an additional focus on the influence of DIMc. Available milk production data from 2005 to 2022 were available for 23,908 cows from 87 herds located throughout the Netherlands and Belgium. Persistency was measured by a lactation curve characteristic decay, representing the time taken to halve milk production after peak yield. Decay was calculated for 8 DPC (0, 30, 60, 90, 120, 150, 180 and 210 d after DIMc) and served as the dependent variable. Independent variables included DPC, DIMc (< = 60, 61-90, 91-120, 121-150, 151-180, 181-210, > 210), parity group, DPC × parity group, DPC × DIMc and variables from 30 d before DIMc as covariates. The results showed an increase in decay, i.e., a decrease in persistency, during pregnancy for both parity groups, albeit in different ways. Specifically, from DPC 150 to DPC 210, multiparous cows showed a higher decline in persistency compared with primiparous cows. Furthermore, a later DIMc (cows conceiving later) was associated with higher persistency. Except for the early DIMc groups (DIMc < 90), DIMc does not impact the change in persistency by gestation. The findings from this study contribute to a better understanding of how DPC and DIMc during lactation influence lactation persistency, enabling more informed decision-making by farmers who wish to take persistency into account in their reproduction management.

Reproductive hormone use and its association with herd-level factors on Dutch dairy farms.

We studied the use of the 3 commonly used reproductive hormones, namely prostaglandins, GnRH, and progesterone, and associated herd-level factors on 760 Dutch dairy farms from 5 veterinary clinics. From 2017 to 2019 we collected data on the sales of reproductive hormones, converted this data into the number of reproductive hormone doses conducted, and expressed this as the annual number of reproductive hormone doses per 100 adult dairy cows. Additional herd-level information was available for 2019. Due to the excess of zeros in the data set (i.e., a substantial number of farms did not use any hormones), we used a zero-inflated negative binomial model to identify related herd-level factors for the use of reproductive hormones. In the entire study period of 2017 to 2019, 5.8% of the dairy farms did not use any reproductive hormones, with the proportion of nonusers varying between 0.0 and 10.3% per veterinary clinic. This proportion was around 13.5% on an annual basis. Prostaglandins were the most frequently used reproductive hormone in Dutch dairy cows (62.9%), followed by GnRH (33.1%) and progesterone (4.0%). Furthermore, participating in a veterinary herd health management program had a significant effect on reproductive hormone use. These farms used more reproductive hormones than farms that did not participate in a herd health management program and were less represented in the group of nonuser farms. Technologies, such as pedometers and automatic milking systems, also had an effect on reproductive hormone use. The presence of pedometers or activity monitors did not reduce the use of the reproductive hormones but was associated with a greater frequency of users. Farms with an automatic milking system used more reproductive hormones than farms with a conventional milking system. With this study, we have made a first step in achieving transparency in the Dutch dairy industry by providing an objective overview of reproductive hormone use on Dutch dairy farms and identifying associations with some herd-level factors.

In vitro maturation of class I oocytes of bovine during different tropical seasons.

The aims of this study were to determine the effects of seasonality on yields and quality of bovine abattoir-derived oocytes and their in vitro maturation (IVM) competence in Thailand, a tropical country. Ovaries were collected monthly from a slaughterhouse, from January to June 2017, separated into cool (January to February), summer (March to April), and rainy (May to June) seasons. Cumulus oocyte complexes (COCs) were obtained by follicular aspiration, and subsequently, oocyte yield and quality were examined. Selected class I and II COCs were cultured under IVM condition, and their maturation capacity was also evaluated. Results showed that average numbers of COCs and class I COCs per ovary were highest in summer. However, no significant seasonal difference in oocyte maturation rates was found. The higher numbers of COCs and class I COCs in summer might be caused by their follicular development during the cool season, and might be a reason why the pregnancy rate in summer is better than in the rainy season.

Cysteamine supplementation during in vitro maturation of slaughterhouse- and opu-derived bovine oocytes improves embryonic development without affecting cryotolerance, pregnancy rate, and calf characteristics.

Optimization of ovum pick up (OPU) followed by in vitro embryo production (IVP) is strongly driven by the needs of both beef and dairy cattle breeders to enhance genetic improvement. The rapidly growing use of genomic selection in cattle has increased the interest in using OPU-IVP technology to increase the number of embryos and offspring per donor, thus allowing enhanced selection intensity for the next generation. The aim of this study was to optimize embryo production through supplementation of cysteamine during in vitro maturation (IVM) and in vitro culture (IVC) of both slaughterhouse- and OPU-derived oocytes. The effects on embryo production and on embryo cryotolerance, post-transfer embryo survival, and calf characteristics, including gestation length, birth weight, perinatal mortality, and sex ratio were studied. In study 1, immature slaughterhouse-derived cumulus-oocyte complexes (COCs) were matured in IVM medium supplemented with or without 0.1 mM cysteamine, fertilized and cultured for 7 days in 0.5 ml SOFaaBSA. In study 2, cysteamine was present during both IVM (0.1 mM) and IVC (0.01, 0.05, 0.1 mM) from Days 1 to 4. In study 3, OPU-derived COCs were matured in medium supplemented with or without 0.1 mM cysteamine in a 2 × 2 factorial design (OPU week and cysteamine treatment). Embryos were evaluated for stage and grade on Day 7 and, depending on the number of transferable embryos and recipients available, the embryos were transferred either fresh or frozen-thawed at a later date. The presence of cysteamine during IVM significantly increased the embryo production rate with slaughterhouse-derived COCs (24.0% vs. 19.4%). The higher number of embryos at Day 7 was due to an increased number of blastocysts, whereas the distribution of embryos among different quality grades and cryotolerance was not affected. Embryo production rate was negatively affected when cysteamine was present during both the processes of IVM and IVC during Days 1 to 4 of culture (13.2%-19.3% vs. 26.4%). The presence of cysteamine during IVM of OPU-derived COCs also significantly increased the embryo production rate (34.4% vs. 23.4%). The higher number of embryos was again totally due to an increased number of blastocysts, whereas cryotolerance was not affected. The relative increase in embryo production rate was higher with OPU-derived oocytes compared with slaughterhouse-derived COCs (47% vs. 24%). This improvement resulted in a mean of 1.73 transferable embryos per OPU session compared with 1.06 in the absence of cysteamine. The presence of cysteamine did not affect pregnancy rate, gestation length, birth weight, perinatal mortality, and sex of calves born from either fresh or frozen-thawed embryos. This study reported that cysteamine supplementation during IVM greatly improved the efficiency and affectivity of an OPU-IVP program.

Characterisation of pregnancy losses after embryo transfer by measuring plasma progesterone and bovine pregnancy-associated glycoprotein-1 concentrations.

The aim of this analysis was to determine whether pregnancy loss (PL) after embryo transfer (ET) in cattle was related to maternal progesterone (P4) concentrations during and shortly after ET, and maternal bovine pregnancy-associated glycoprotein-1 (bPAG-1) concentrations in plasma at days 25-35 of gestation. Embryos (n=260) were produced either in vivo after superovulation (n=115), or in vitro from oocytes (obtained with ovum pick-up) in co-culture (n=44) or cultured in a synthetic medium (n=101). Overall, PL was 56.9% (148) and no significant differences occurred in calving rate among the three embryo production groups. There was no difference in P4 concentrations on days 7-14 of gestation in the three groups, nor between ongoing and interrupted pregnancies. Between days 25 and 35 of pregnancy, bPAG-1 concentrations were unaffected by embryo production, but in cattle that had PL between days 26 and 120, four bPAG-1 profiles could be detected. Between days 25 and 32, bPAG-1 concentrations were influenced by PL, and concentrations were significantly lower in animals in which PL occurred between days 26 and 120 than in those animals that aborted later or calved at term. Early P4 concentrations suggested that maternal luteal factors were not responsible for PL which appeared to be caused by impaired conceptus development (regardless of embryo type) as reflected by low maternal bPAG-1 concentrations prior to embryonic death.

Bovine OPU-derived oocytes can be matured in vitro for 16-28 h with similar developmental capacity.

The aim of this study was to determine the optimal maturation culture period of ovum pick up (OPU)-derived cumulus oocytes complexes (COCs) in relation to their developmental capacity. Embryo production, embryo cryotolerance, post-transfer embryonic survival and calf characteristics such as gestation length, birthweight and sex ratio were investigated. This retrospective study covers the analyses of ovum pick up -in vitro production and calving results from a commercial programme that took place between March 1994 and September 2004. Donors were both heifers (of which approximately 90% pregnant) and cows (of which approximately 10% pregnant). Embryo production analyses were based on 7800 OPU sessions conducted from January 1995 until January 1999. Analyses of calving rate were based on 13 468 embryo transfers performed during January 1995 until May 2002. Analyses on calf characteristics were based on 2162 calves born between March 1994 and September 2004. The in vitro maturation culture period ranged from 16 to 28 h. The mean production rate of transferable embryos was 16.5% (1.2 embryos per OPU session). Length of maturation culture period did not affect the production of transferable embryos. Mean calving rate was 40.9% and 38.7% for fresh and frozen/thawed embryos, respectively. Calving rate was not affected by the maturation culture period. Mean birthweight, gestation length and proportion of male calves were 46 kg, 281.9 days and 52.8%, respectively. Maturation culture period did not affect these variables. In conclusion, this study shows that the in vitro maturation culture period within the range of 16-28 h does not affect in vitro embryo production, embryo cryotolerance, post-transfer embryonic survival and calf characteristics, suggesting that all COC batches collected by OPU on the same day, can be fertilized in one IVF session without a significant loss in the production from oocyte to calf.

Effects of superovulation with oFSH and norgestomet/GnRH-controlled release of the LH surge on hormone concentrations, and yield of oocytes and embryos at specific developmental stages.

The objective of this study was to evaluate a new superovulation procedure with oFSH after temporary suppression of the endogenous LH surge by norgestomet followed by administration of GnRH, to collect bovine oocytes and embryos at specific developmental stages. Since 1999, our research group applies this superovulation procedure with controlled release of the endogenous LH surge. The objective of this study is to verify if this procedure is reliable for collection of oocytes and embryos at specific time points of development and if it produces a sufficient number of both oocytes and embryos of good quality. This procedure was validated regarding to hormonal characteristics, superovulatory response and both oocyte and embryo yield at different times of in vivo development. The results demonstrate that the procedure used to control the occurrence of the pre-ovulatory LH surge was effective in 92% of the animals (n = 238) and even in 99% of the animals the oocytes and embryos were collected at the intended stage of development. The superovulatory response and both oocyte, embryo yield and quality were similar to the average yield in Europe reported by Association Européenne de transfert embryonnaire (AETE). In conclusion, this superovulation procedure provides a valid tool to collect oocytes and embryos at specific time points of development.

Analysis of the economically optimal voluntary waiting period for first insemination.

The voluntary waiting period (VWP) is defined as the time between parturition and the time at which the cow is first eligible for insemination. Determining the optimal VWP from field data is difficult and unlikely to happen. Therefore, a Monte-Carlo dynamic-stochastic simulation model was created to calculate the economic effects of different VWP. The model is dynamic and uses time steps of 1 wk to simulate the reproductive cycle (ovulation, estrous detection, and conception), the occurrence of postpartum disorders, and the lactation curve. Inputs of the model were chosen to reflect the situation of Dutch dairy cows. In the model, we initially created a cow of a randomly selected breed, parity, month of calving, calf status of last calving, and expected 305-d milk yield. The randomly varied variables were based upon relevant distributions and adjusted for cow statuses. The lactation curve was modeled by Wood's function. The economic input values in the analysis included: cost of milk production (€0.07 to €0.20 per kg), calf price (€35 to €150 per calf), AI cost (€7 to €24 per AI), calving management cost (€137 to €167 per calving), and culling cost, expressed as the retention pay-off (€118 to €1,117). A partial budget approach was used to calculate the economic effect of varying the VWP from 7 to 15 wk postpartum, using a VWP of 6 wk as reference. Per iteration, the VWP with either the lowest economic loss or the maximum profit was determined as the optimal VWP. The optimal VWP of most cows (90%) was less than 10 wk. On average, every VWP longer than 6 wk gave economic losses. Longer VWP were in particular optimal for the first parity of breeds other than Holstein-Friesian, cows calving in winter with low milk production, high milk persistency, delayed peak milk yield time, a delayed time of first ovulation, or occurrence of a postpartum disorder, and while costs of milk production are low and costs for AI are high.

Chemotactic activity of cotyledons for mononuclear leukocytes related to occurrence of retained placenta in dexamethasone induced parturition in cattle.

Induction of parturition with glucocorticosteroids in cattle is used for research purposes, in diseased or injured pregnant cows, and as a management tool to time parturition. A negative side effect of induction of parturition with glucocorticosteroids is the high incidence of retained placenta that occurs after these calvings. Reaction of the maternal immune system against the 'foreign' foetal membranes contributes to the breakdown of the foetal-maternal attachment. Several studies indicate that failure of this immune assisted detachment increases the occurrence of retained placenta. We hypothesized that retained placenta occurring after induction of parturition with glucocorticosteroids is caused by failure of immune assisted detachment of the foetal membranes. The chemotactic activity of cotyledons for mononuclear leukocytes was used as a parameter to see whether immune assisted detachment of the foetal membranes had occurred. Cotyledons were collected from spontaneously calving non-retained placenta cows and from dexamethasone induced non-retained placenta and retained placenta cows. The study showed that the chemotactic activity of cotyledons for mononuclear leukocytes was lower (P < 0.001) in cotyledons obtained from retained placenta cows in which parturition was induced with dexamethasone compared to the chemotactic activity of cotyledons obtained from spontaneously calving non-retained placenta cows, whereas the chemotactic activity of cotyledons obtained from induced non-retained placenta cows was not lower (P = 0.10) than the chemotactic activity of cotyledons obtained from spontaneously calving non-retained placenta cows. We concluded that induction of parturition with dexamethasone causes a failure of immune assisted detachment of the foetal membranes and the accompanying release of chemotactic factors. As a result, the chemotactic activity of cotyledons for mononuclear leukocytes is lower in induced retained placenta cows than in cotyledons from non-retained placenta cows in which successful immune assisted detachment of the foetal membranes occurs.

Cow effects and estimation of success of first and following inseminations in Dutch dairy cows.

The objective of this research was to determine the contribution of cow factors to the probability of successful insemination accounting for the serial number of inseminations in analysis. The investigation was performed with 101,297 insemination records in 51,525 lactations of different cows from 1368 herds obtained from the Dutch milk production recording database. Cows that had a first insemination (AI) between 40 and 150 days post-partum with one or more inseminations (≤6 inseminations) were selected. An insemination was defined successful when not followed by another insemination and when the cow calved between 267 and 295 days after insemination, or when the cow was culled between 135 and 295 days after the last insemination. Breed, parity, days in milk, lactation curve characteristics, milk production traits, moment of AI related to peak milk yield time (before or after peak milk yield), the last calf (female, male, twin or stillbirth) and season of insemination were selected as independent parameters for a model with successful rate of insemination as dependent parameter. A multivariable logistic regression model was used within cow and farm as a random effect. The probability of successful insemination was the highest in the first insemination and decreased in the following inseminations. However, the success rate of all inseminations increased in a later stage of lactation. The improvement in the successful inseminations in a later stage of lactation was better in multiparous cows than in first parity cows. Insemination in summer and before peak milk yield time reduced the success of insemination. The success rate was the lowest in 100% Holstein Friesian cows compared with other breeds and was the highest when the last calf was a female calf compared to a male calf, twin or stillbirth. In conclusion, the success of first and following inseminations depended on parity, breed, season of insemination, last calf status, daily milk yield at insemination date, serial insemination number and days in milk at insemination date.

Effect of milk yield characteristics, breed, and parity on success of the first insemination in Dutch dairy cows.

The objective of this study was to determine the contribution of cow factors to the probability of a successful first insemination (SFI). The investigation was performed with 51,791 lactations from 1,396 herds obtained from the Dutch dairy cow database of the Cattle Improvement Co-operative (CRV). Cows that had the first insemination (AI) between 40 and 150 d postpartum were selected. The first AI was classified as successful when cows were not reinseminated and either calved between 267 and 295 d later or were culled within 135 to 295 d after first AI. The lactation curve characteristics of individual lactations were estimated by Wilmink's curve using the test-day milk records from CRV. The lactation curve characteristics (peak milk yield, milk yield at the first-AI date, time of peak yield (PT), and milk persistency) were calculated. Breed, parity, interval from calving to first AI (CFI), lactation curve characteristics, milk production traits, moment of AI related to PT (before or after PT), calf status, month of AI, and month of calving were selected as independent variables for a model with SFI as a dependent variable. A multivariable logistic regression model was used with farm as a random effect. Overall SFI was 44%. The effect of parity on SFI depended on CFI. The first-parity cows had the greatest SFI (0.43) compared with other parities (0.32-0.39) at the same period of CFI before 60 d in milk (DIM), and cows in parity ≥5 had the least SFI (0.38-0.40) when AI was after 60 DIM. After 60 DIM, extending CFI did not improve SFI in the first-parity cows, but SFI was improved in multiparous cows. Holstein-Friesian cows had lesser SFI (0.37) compared with cross-breed cows (0.39-0.46). Twin and stillbirth calving reduced SFI (0.39) compared with a single female calf (0.45) or a male calf (0.43) calving. The SFI in different months of AI varied and depended on CFI. Cows that received AI before 60 DIM had a lesser SFI, especially in March, June, and July (0.18, 0.35, and 0.34, respectively). Artificial insemination before PT reduced SFI (0.39) in comparison with AI after PT (0.44). The effect of milk yield at the first-AI date on SFI varied depending on CFI. After 60 DIM at the same period of CFI, a high level of milk yield at the first-AI date reduced SFI. In conclusion, knowledge of the contribution of cow factors on SFI can be applied to support decision making on the moment of insemination of an individual cow in estrus.

Economic consequences of reproductive performance in dairy cattle.

The net economic value of reproductive efficiency in dairy cattle was estimated using a stochastic dynamic simulation model. The objective was to compare the economic consequences of reproductive performance scenarios ("average" and "poor") of a cow having a good reproductive performance and to explore which reproductive factors have an important impact on economic efficiency. A "good" reproductive performance scenario was defined with 1 ovulation rate (POVU(i)), 0.7 estrus detection rate (PEst), 0.7 conception rate (PCon), 0.03 incidence rate of postpartum disorders prolonging the ovarian cyclicity (CO), 0.2 incidence rate of postpartum disorders reducing conception (ME), 0.05 embryonic death rate (ED), and voluntary waiting period (VWP) of 9 wks pp (post partum). In the current situation of dairy cows in the Netherlands, an "average" reproductive scenario (0.95 POVU(i), 0.5 PEst, 0.5 Pcon, 0.07 CO, 0.27 ME, 0.07 ED and VWP of 12 wks pp) and a "poor" reproductive scenario (0.90 POVU(i), 0.3 PEst, 0.3 Pcon, 0.11 CO, 0.33 ME, 0.09 ED and VWP of 15 wks pp) were identified. A sensitivity analysis was performed by comparing changes of single effect of factors in a good and poor scenario with the average scenario. The mean net economic loss (NEL(i)) compared with the good scenario was euro 34 and euro 231 per cow per year for the average and poor reproductive performance scenario, respectively. Increasing the calving interval resulted in greater economic loss. The important factors on the cost of reproductive efficiency were the involuntary culling cost and the return of milk production. Variation in PCon, PEst, ME, ED, and VWP had large impacts on economic benefits.

Postpartum anestrus in dairy cattle.

Fertility of the postpartum period is negatively influenced by the incidence of anestrus. The latter condition is characterized by the absence of estrous behavior, which may be an indication of suboptimal conditions (e.g., inadequate peripartum nutrition) or pathologic conditions (e.g., chronic debilitating diseases or uterine and ovarian diseases). Although initiation of ovarian follicular growth in the postpartum period is generally not affected, subsequent development (deviation) and the fate of the dominant follicle are the primary factors that affect reestablishment of ovarian cyclicity. Anestrus can be classified based on the three functional states of follicular development; that is, follicle emergence, deviation, and ovulation. Prevention of anestrus is preferable to treatment and can be achieved in part by maintaining a healthy periparturient period. To better understand the etiology of anestrus and its prevention, research is urgently needed in the following three areas: the role of peripartum disease conditions that influence reproduction, genes involved in ovulation, and the influence of proteins (e.g., leptin) that appear to be important links between metabolic signals and the neuroendocrine axis.

Veterinary advice for entrepreneurial Dutch dairy farmers: from curative practice to coach-consultant: what needs to be changed?

Dairy farms are tending to become larger, with a milk quota of more than 8 tons a year, and are managed by entrepreneurial dairy farmers with their own specific characteristics and farming style. Some Dutch veterinary practices appear unable to respond to this different style and often do not serve such farms or lose them as client. Moreover, the veterinary curriculum often focuses on traditional, family-run, smaller dairy operations and not on larger farms, which raises the question whether newly qualified veterinary practitioners are adequately trained to provide these entrepreneurial farmers with the services they require. This article addresses the characteristics of entrepreneurial dairy farmers and those of cattle practitioners, to determine whether cattle practitioners need to acquire other skills to better prepare them for their coaching-consultant tasks on larger dairy farms.

[Transvaginal aspiration as first treatment of ovarian follicular cysts in dairy cattle under field circumstances]. / Transvaginale aspiratie als primaire behandeling van ovariële follikelcysten bij melkkoeien onder praktijkomstandigheden.

On dairy farms covered by the ambulatory service of the Department of Farm Animal Health, 55 Holstein-Friesian cows with suspected ovarian follicular cysts on the basis of rectal palpation underwent confirmatory transrectal ultrasonography. An ovarian follicular cyst was confirmed in 28 cows (50.9%) and was treated by single transvaginal-guided needle aspiration. Cows with a corpus luteum or a luteinized cyst were not treated. After aspiration, 82.1% (n=23) of the cows showed oestrus behaviour at 13.3 +/- 6.0 days. Artificial insemination was performed during the first heat after cyst aspiration and resulted in a pregnancy rate of 64.2%. In conclusion, single transvaginal-guided needle aspiration of ovarian follicular cysts is an easy and good method for the treatment of follicular cysts. Moreover, it is a safe and good alternative method for the manual, active rupturing of cysts during rectal palpation.

Plasma concentrations of bovine pregnancy-associated glycoprotein (bPAG) do not differ during the first 119 days between ongoing pregnancies derived by transfer of in vivo and in vitro produced embryos.

Calves derived from IVP embryos may suffer from the large offspring syndrome that has been related to effects of in vitro culture on the intrinsic quality of the embryo. Limited information is available on the role of the placenta in such cases. In this study, bovine pregnancy-associated glycoprotein (bPAG) was used as a marker to test whether placental function is influenced by the route of embryo production. Therefore, from day 7 until day 119 of ongoing gestations, resulting from transfer of MOET (n = 53), IVP-co-culture (n = 21) and IVP-SOF (n = 38) embryos, bPAG levels were compared in peripheral plasma of recipients. Plasma progesterone levels were compared as well. From day 25 of gestation onwards, bPAG could be detected in all recipients and the levels were significantly influenced by the day of gestation. Although IVP calves were significantly heavier than the in vivo produced calves, this difference was not reflected in the bPAG profiles of the embryo production groups. Yet, the mean bPAG level of the three last sampling moments (days 105-119) tended to be positively related to the birth weight of the calves, irrespective of the embryo production technique. Progesterone concentrations were not influenced by route of embryo production, but were significantly affected by parity of the recipient and day of gestation.

Effects of non-esterified fatty acids on bovine granulosa cells and developmental potential of oocytes in vitro.

High yielding dairy cows experience a negative energy balance (NEB) shortly after parturition, which is accompanied by high concentrations of non-esterified fatty acids (NEFA) in blood up to approximately 3 weeks post partum. We hypothesized that the elevated plasma NEFA concentration causes lower fertility by exerting negative effects on granulosa cells and oocytes in the ovary, leading to less viable embryos and insufficient corpora lutea. In two series of experiments, we studied the effects of a realistic NEFA (C18:1) concentration on both the proliferation and the progesterone production of follicular granulosa cells in vitro (part I) and on maturation, fertilization and developmental potential of oocytes (part II). For part I, granulosa cells were added to 4 groups of dishes with four different media and cultured for nine consecutive days. After a preculture period of 42h, the presence of NEFA had a negative effect on the proliferation of granulosa cells. No effect of NEFA on the amount of progesterone production per cell was observed. For part II, a total of 1804 cumulus-oocyte-complexes were collected from slaughterhouse ovaries. Using a subgroup of 690 COC, maturation medium with NEFA caused a delay in maturation. Using another 1114 COC, fertilization, cleavage, and embryonic development after maturation in presence of NEFA were significantly reduced. We concluded that the presence of NEFA in follicular fluid and blood of post partum cows may reduce fertility due to hampered embryonic development and subnormal CL function.