Biomedical subjects

T A Kruip

Publications and source records attributed to T A Kruip.

At least 19 recordsLinked to original sources

PubMed2001

Bovine cumulus-oocyte-complex-quality is reflected in sensitivity for alpha-amanitin, oocyte-diameter and developmental capacity.

The aim of the present study was to find more parameters to define developmental competence of cumulus-oocyte-complexes (COCs). Bovine COCs were divided into five groups based on their morphology. In order of increasing level of atresia: COC-A had a bright, compact cumulus investment; COC-B1 also had a compact cumulus investment, but was darker than COC-A; the color of COC-B2 was comparable with COC-B1 but the corona radiata appeared to dissociate from the rest of the cumulus investment; the cumulus of COC-B3 was almost black and the corona radiata was almost completely dissociated from the rest of the cumulus investment; COC-C had a strongly expanded cumulus investment with dark spots of degenerated cells. An increasing level of atresia was accompanied by: (1) an increasing zona pellucida diameter (147.6, 150.8, 151.0, 154.3 and 155.1 microm, respectively, for COC-A, COC-B1, COC-B2, COC-B3 and COC-C); (2) an increasing oocyte diameter except for COC-C (120.9, 122.8, 122.8, 123.9 and 118.4 microm, respectively); (3) an increasing developmental competence except for COC-C (13.9, 14.7, 17.4, 19.1 and 11.5%, respectively, development to morula and blastocyst after in vitro embryo production (IVP)) and (4) by a increasing percentage of oocytes exhibiting germinal vesicle breakdown (GVBD) after 24h culture with alpha-amanitin and FSH, except for COC-A (34.4, 68.6, 51.0, 22.0 and 4.8%, respectively, oocytes arrested in GV). In general, embryo-quality, expressed in nuclei-count, was significantly affected by COC-quality (P < 0.05) with B3 > B1> B2 > A > C. However, if the developmental stages were compared separately, this effect was less evident. It was found that, in absence of FSH, alpha-amanitin was unable to inhibit GVBD and that the success of GVBD inhibition was positively correlated to the amount of cumulus cells surrounding the oocyte. COCs that had been exposed to alpha-amanitin and FSH during maturation retained the ability to cleave after IVF, but were unable to develop any further.

Amanitins↗

PubMed2001

Effect of urea during in vitro maturation on nuclear maturation and embryo development of bovine cumulus-oocyte-complexes.

High concentrations of urea in reproductive tract fluids are detrimental to bovine reproduction. Therefore, in experiment 1, the effect of 6 mM urea on nuclear maturation of cumulus-oocyte-complexes (COC) collected from abattoir ovaries was studied. After 4, 8, 12, 16, 20, and 24 h of in vitro maturation, the nuclear stages of samples of the COC were determined. During the first 8 h of maturation, germinal vesicle breakdown and chromosome condensation, resulting in the metaphase I stage, occurred at higher rates in the presence of urea. Segregation of the chromatids and extrusion of the polar body seemed to be impaired in the presence of urea, resulting in a higher percentage of oocytes arrested in metaphase I or telophase, and a lower percentage of oocytes in metaphase II after 24 h of maturation. Overall, nuclear progression of COC matured in the presence of urea differed from COC matured in control medium. In experiment 2, COC were matured for 24 h either in the presence or absence of 6 mM urea followed by in vitro fertilization and culture. After fertilization, a sample of the COC was fixed and stained to determine the fertilization rate. The cleavage rate was determined 3 d after start of maturation, and the stage of embryonic development was recorded 7 and 9 d after start of maturation. Based on cultured oocytes, urea in the maturation medium decreased the subsequent percentage of fertilization, cleavage, and development on d 7 and 9 (43.2, 56.1, 14.8, and 18.2%, respectively for urea vs. 64.1, 68.8, 22.4, and 23.9%, respectively for the control group). Embryonic development as a percentage of cleaved oocytes was not significantly affected by urea. Therefore, negative effects of urea were evident primarily during oocyte maturation and fertilization.

Animals↗

PubMed2000

Altered functional and immunophenotypical properties of neutrophilic granulocytes in postpartum cows associated with fatty liver.

The intention of the study was to analyze the relationship between liver triacyl glycerol content (liver TAG content) and immunophenotypical and functional properties of polymorphonuclear neutrophilic granulocytes (PMN) of dairy cows in the peripartum period. We investigated characteristics of bovine PMN from the blood and uterus of clinically healthy cows in the periparturient period. The numbers of circulating leukocytes and segmented granulocytes continuously increased until parturition and declined afterwards to starting values. This was independent of the liver TAG content and mainly affected neutrophils. The liver TAG content exceeded 40 mg/g liver, the reference value, in 12 of 19 cows in the first two weeks postpartum. Increased liver TAG content, > 40 mg/g, went in parallel with a reduced expression of function-associated surface molecules on blood neutrophils (e.g. CD11b/CD18 = CR3 and CD11c/CD18 = CR4). Moreover, in cows with high liver TAG levels the antibody-independent and -dependent cellular cytotoxicity (AICC, ADCC) of blood PMN was markedly reduced. PMN also were less capable of ROS generation after stimulation with Phorbol Myristate Acetate (PMA). In comparison with contemporarily harvested blood PMN, neutrophils recovered from the uterine lumen showed a decreased expression of 4/6 examined surface structures. Only the expression densities of CR3 molecules and those detected by mAb IL-A110 were enhanced on uterine PMN. The cytotoxic capacity and the ROS generation were significantly lower for uterine PMN than for blood PMN. The results suggest that increased liver TAG content in the first and second week after calving is associated with decreased functional capacities of PMN derived from blood and uterus. This may help to explain why cows who are too fat at calving (who therefore have an increased liver TAG content) have a higher incidence of infectious diseases such as endometritis

3-Hydroxybutyric Acid↗

PubMed2000

Robot milking and effect on reproduction in dairy cows: a preliminary study.

The aim of this study was to analyse whether automatic milking has an effect on reproduction in dairy cows. Probably the most important consequence of automatic milking is an increased milking frequency. More frequent milking means more production and thereby an extra threat for the energy balance during the first months of lactation. A deep negative energy balance (NEB) is often followed by negative effects on fertility. However, tested under experimental circumstances, automatic milking with a frequency not higher than 3 times/day and with an individual management and feeding system has benefits for production and very little negative effects on reproduction. Experimental farms with automatic milking systems have fertility figures that do not differ from experimental farms with conventional milking systems. Additional and more data of automatic milking under farm conditions has to be collected and evaluated before the effect on fertility can be assessed with more certainty.

Animal Nutritional Physiological Phenomena↗

PubMed2000

Environment of oocyte and embryo determines health of IVP offspring.

In vitro embryo production (IVP) enhances the number of offspring from a single female and offers the possibility of accelerated genetic progress in animal husbandry. However, it also leads to a low but unacceptable percentage of anomalies in the offspring. The aim of this paper is to introduce the three speakers at this afternoon session who will speak about the demands of culture conditions and the endometrial environment to support normal embryonic development without effects on the embryonic genome. It will be argued that the in vitro conditions should mimic precisely the oviductal contributions to homeostatic mechanisms within the embryo. The further normal development can be guaranteed at synchrony in development of both endometrium and embryo. If that is not the case one can expect disturbances of gene expression, in particular of imprinted genes. However, it cannot be excluded that some processes might have started already in the cytoplasm of the oocyte. Since the oocyte was not planned to be a separate subject in this symposium, this introduction is also aimed to ask attention for the selection of cumulus-oocyte-complexes (COCs) and the conditions around oocyte maturation in vitro. The optimal quality of both the oocyte and maturation medium are prerequisites for an undisturbed cytoplasmic maturation. It has been argued that the exclusion of COCs from atretic follicles, the abjuration of the use of serum and high O2 tension in the gas phase might help to reduce the proportion anomalies in the offspring after synchronous transfers. In human IVF, in vivo matured oocytes are used with no great problems. But before IVP, including oocyte maturation in vitro (IVM) and a longer lasting embryo culture (IVC), will be introduced into the human assisted reproduction, it is important to think about the ins and outs of the potential causes for deviations.

Animals↗

PubMed2000

Relationship among estradiol, cortisol and intensity of estrous behavior in dairy cattle.

Economic profitability of a dairy farm is based, in part, on the calving interval of the cows. The optimal interval is 365 d. To achieve this, the cow needs to be pregnant within 85 d post partum. The first and most problematic step in this process is the determination of the optimal time for insemination, which is based on estrous behavior. The expression of estrous behavior, however, is at a low level in modem dairy herds, resulting in low detection rates and longer calving intervals. In the present study, a point scale was used to monitor postpartum, nonpregnant cows for estrous symptoms. Frequent blood samples were taken around estrus, and the cows were fit with pedometers to measure their activity. Correlations between the occurrence of symptoms of estrus and levels of estradiol and cortisol were then analyzed. Standing heat, the standard symptom of estrus, was observed in only 53% of the cows. A high correlation of 0.7 was found between estradiol concentration and estrous behavior. This was empasized by the fact that the estradiol level reached its highest level of 7.76 + 2.39 (SD) pg/mL at the same time as the highest behavior score. The highest pedometer readout lagged 8 h behind this moment. Cortisol levels did not exceed the physiological levels in rest situations but showed an increase at the time estrous behavior was at its maximum. The present study showed that standing heat is not the primary symptom for detecting estrus in cows. Pedometers are a useful aid but they have to be read several times a day. The high correlation between the visual symptoms of estrus and estradiol concentrations indicates that visual estrus detection is an efficient, reliable way to determine the right time for insemination.

Animals↗

PubMed2000

Effect of ovarian phase and follicle quality on morphology and developmental capacity of the bovine cumulus-oocyte complex.

In order to study the effect of the follicular environment on the quality and developmental competence of cumulus-oocyte complexes (COC), COC were collected from nonatretic (NA), light atretic (LA), atretic (A), and heavily atretic (HA) follicles. Cumulus-oocyte complexes were also collected from early-luteal phase ovaries (EL), from late-luteal phase ovaries (LL), from follicular phase ovaries (F), and from ovaries from non-cyclic animals (NC). Each COC was assigned to one of three quality groups based on the appearance of the cumulus investment: 1) COC-A: compact and bright, 2) COC-B: less compact and dark, and 3) COC-C: strongly expanded cumulus with dark spots. There was a high correlation between follicle quality and the distribution of the COC over the three COC qualities. The COC-A were mainly but not exclusively derived from NA follicles, COC-B were mainly derived from all classes of atretic follicles, and COC-C nearly exclusively originated from HA follicles. The developmental capacity of COC-A and COC-B, which was measured by in vitro embryo production, was consistent over the follicle qualities, except for COC-B obtained from HA follicles. They showed lower development (10.6%, P < .05) compared with COC-B from the other follicle qualities (18.9, 18.7, and 19.8%, respectively, for COC-B from NA, A, and LA follicles). The COC-B from atretic follicles produced more blastocysts (19.8%) than COC-A (12.7%, P < .05). The overall percentage of produced embryos per follicle class seemed to increase with increasing signs of atresia, except if the COC were derived from HA follicles. This increased percentage of embryos was, however, not due to a better quality of COC, but to a higher percentage of COC-B coming from these follicles. The stage of the cycle had no effect on the distribution of the COC over the three COC qualities or on the developmental capacity of COC-A or COC-B, except for COC-A from EL ovaries, which produced more (P < .05) blastocysts than COC-A from the other luteal phases (12.5% vs approximately 8%). A follow-up study was performed trying to elucidate why COC-B possess a higher developmental potency than COC-A. The answer was sought in the oocyte maturation. At several time points during maturation, oocytes were evaluated for their nuclear stage. At all time points COC-B seemed to be a few hours ahead of COC-A, and after 24 h of maturation more COC-B had reached the metaphase-2 stage. This might mean that COC-A need a longer maturation period than COC-B.

Animals↗

PubMed2000

Interrelation of fatty acid composition in adipose tissue, serum, and liver of dairy cows during the development of fatty liver postpartum.

We investigated the composition of fatty acids in adipose tissue, serum, and liver of cows that were fed at restricted energy intake or were overfed during the dry period. Overfed cows had higher concentrations of serum nonesterified fatty acids and consequently accumulated greater amounts of triacylglycerols in the liver than did cows that were fed at restricted energy intake. The percentages of the different fatty acids present in adipose tissue were similar for both groups and did not change during sampling intervals. Before parturition, concentrations of the individual fatty acids present in serum were similar between groups. After parturition, concentrations of major fatty acids in serum, including palmitic, stearic, oleic, and linoleic acids significantly increased in both groups and were higher in overfed cows than in cows that were fed at restricted energy intake. The shift of concentrations of the different fatty acids present in the liver--as a result of increased lipolysis-was observed in palmitic, oleic, and linoleic acids but not stearic acid, suggesting that stearic acid is used by the liver (i.e., oxidation) or is considerably secreted through the milk, thereby not increasing in accumulation in the liver. In conclusion, different feeding regimens during the dry period do not influence the composition of fatty acids in adipose tissue. More intensive lipolysis results in increased concentrations of palmitic, stearic, oleic, and linoleic acids in the blood; subsequently, these fatty acids, excluding stearic acid, greatly accumulated in the liver.

Adipose Tissue↗

PubMed1999

[Nutrition and fertility in agricultural domestic animals].

This paper deals with the reduction of fertility in cattle and pigs due to increased production. Special attention has been paid to the negative energy and protein balances during the first weeks post partum. It has been questioned which organs and organ systems become affected by the changes in these balances. For that study, the animal model of the overconditioned cow ante partum has been developed and used. The paper is dealing with possible effects on the hypothalamic-hypophyseal (HH) system and LH release, on the ovaries with secretion of steroids and release of properly matured oocytes, on the uterus as milieu for the embryo and on the immune system with leucocytes that play a role in follicle development and ovulation. The preliminary results of the first experiments open the discussion whether the reduction in fertility in this model is the consequence of the direct effect of glucose and insulin on the HH system and ovaries or of the indirect toxic effect of the non-esterified fatty acids, resulting from a temporary decrease of insulin, on granulosa cells and gametes.

Animal Nutritional Physiological Phenomena↗

PubMed1999

Relationship between triacylglycerol concentration in the liver and first ovulation in postpartum dairy cows.

The relationship between accumulation of triacylglycerols in the liver and first ovulation post partum was studied in an experiment with 32 dairy cows. During the dry period, 16 treated cows were fed a high energy diet to induce overconditioning, while 16 control cows were fed a recommended diet. After parturition, the treated cows entered deeper negative energy balance state, required more days to reach the nadir of negative energy balance, required more days to regain positive value of energy balance, and lost more body weight than the control cows. The treated cows also accumulated greater amounts of triacylglycerols in the liver and had higher concentrations of nonesterified fatty acids and 3-hydroxybutyrate post partum than the control cows. Although the number of days to first ovulation did not differ significantly between the 2 groups, the results of pooled data demonstrated that the liver triacylglycerol concentration was positively correlated with the number of days to first ovulation. In conclusion, negative energy balance caused the body fat of cows to mobilize. As a consequence, the cows accumulated greater amounts of triacylglycerols in the liver, and this accumulation was associated with an increase in the interval from parturition to first ovulation.

3-Hydroxybutyric Acid↗

PubMed1999

Pregnancy diagnosis in sows: direct ELISA for estrone in feces and its prospects for an on-farm test, in comparison to ultrasonography.

The usefulness of fecal estrone (E1) measurement as a tool for pregnancy diagnosis was investigated. Concentrations of E1 were measured in feces from pregnant and nonpregnant sows by a direct ELISA without extraction. Highly significant differences in E1 concentrations were found in feces from nonpregnant and pregnant sows (P = 0.016 to < 0.001). Pregnancy diagnosis on Days 26 to 32 after insemination, based both on fecal E1 concentrations as measured by ELISA and ultrasonography using a 5.0 MHz linear-array transducer, was performed in a group of 496 gilts and sows. The fecal E1 test had a sensitivity (correct diagnosis of pregnancy) of 96.5% and a specificity (correct diagnosis of nonpregnancy) of 93.6%, using 3.65 ng E1/g feces as a cut-off value. For ultrasonographic pregnancy diagnosis the test sensitivity and specificity were 99.3 and 92.5%, respectively. Although an increase of fecal E1 concentrations was noticed for increasing litter sizes, the results indicated that these concentrations could not be used to predict litter size. It is concluded that the distribution of fecal E1 concentrations in both nonpregnant and pregnant sows offers a suitable basis for the development of a simple, sow-side pregnancy test.

Animal Husbandry↗

PubMed1999

Relationship between overfeeding and overconditioning in the dry period and the problems of high producing dairy cows during the postparturient period.

In dairy cows, overfeeding during the dry period leads to overcondition at calving and to depression of appetite after calving. As a consequence, at calving overconditioned high-producing dairy cows inevitably go into a more severe negative energy balance (NEB) postpartum than cows that have a normal appetite. During the period of NEB, the energy requirements of the cow are satisfied by lipolysis and proteolysis. Lipolysis results in an increased concentration of non esterified fatty acids (NEFA) in the blood. In the liver, these NEFA are predominantly esterified to triacylglycerols (TAG) that are secreted in very low density lipoproteins (VLDL). In early lactation in cows with a severe NEB, the capacity of the liver to maintain the export of the TAG in the form of VLDL in balance with the hepatic TAG production is not always adequate. As a result, the excess amount of TAG accumulates in the liver, leading to fatty infiltration of the liver (hepatic lipidosis or fatty liver). The NEB and/or fatty liver postpartum are frequently associated with postparturient problems. In general, a severe NEB induces changes in biochemical, endocrinological, and metabolic pathways that are responsible for production, maintenance of health, and reproduction of the postparturient dairy cow. These changes include a decrease in blood glucose and insulin concentrations, and an increase in blood NEFA concentrations. High NEFA concentrations caused by intensive lipolysis are accompanied by impairment of the immune system, making the cows more vulnerable to infections. Metabolic diseases such as ketosis, milk fever, and displaced abomasum are related to overcondition at calving. The changes in biochemical, endocrinological, and metabolic pathways are associated with delay of the first visible signs of oestrus, an increase in the interval from calving to first ovulation, a decrease in conception rate, and a prolonged calving interval. It is possible that the increased blood NEFA concentration directly impairs ovarian function.

Animal Feed↗

PubMed1999

Hepatic fatty acid composition in periparturient dairy cows with fatty liver induced by intake of a high energy diet in the dry period.

The present study compared the hepatic fatty acid composition of cows that were fed a high energy diet during the dry period to induce fatty liver after parturition with that of control cows. Treated cows had higher concentrations of plasma nonesterified fatty acids as a result of greater lipolysis after parturition than did control cows; consequently, the treated cows accumulated greater amounts of triacylglycerols in the liver. Before parturition, treated cows had lower percentages of oleic acid and higher percentages of linoleic acid than did control cows, but percentages of other fatty acids were similar for both groups. After parturition, percentages of each fatty acid were changed substantially, particularly the four major fatty acids (palmitic, stearic, oleic, and linoleic acids). For treated cows, the percentages of palmitic and oleic acids were higher at 0.5 wk after parturition than at 1 wk before parturition; percentages of stearic and linoleic acids decreased. Unlike treated cows, the percentages of both oleic and linoleic acids in the control cows did not change during that time. Moreover, we found that when lipolysis decreased, as indicated by lower plasma nonesterified fatty acid concentrations, the percentages of each fatty acid gradually rebounded toward the concentrations measured before parturition; this observation indicates that the shift in hepatic fatty acid composition is influenced by lipolysis. The increased lipolysis after parturition led to a vast increase in the hepatic triacyglycerol concentration and to a shift in hepatic fatty acid composition.

3-Hydroxybutyric Acid↗

PubMed1997

Follicular dynamics, repeatability and predictability of follicular recruitment in cows undergoing repeated follicular puncture.

The aim of our study was to characterize follicular recruitment and growth in response to the repeated removal of follicles. All tertiary follicles (> 2 mm of diameter) in the ovaries of 10 non-lactating Holstein Friesian cows were punctured at midcycle (Day 0) by means of an ultrasound-guided needle. Puncture sessions were scheduled twice weekly at 3- or 4-d intervals over 3 mo. In the middle of the experiment, i.e., Week 7, the effects of 2- and 5-d intervals between follicular punctures were tested and compared with the previous 3- and 4-d intervals. After this period, 6 animals were slaughtered to study the effect of puncturing on gross ovarian morphology. The protocol of puncturing follicles with 3- to 4-d intervals was continued for an additional 3 mo in the remaining 4 animals. Twice-weekly puncturing of all tertiary follicles abolished estrous cycles and lead to an increase in follicular wave frequency without apparent negative effects on either the reproductive tract or ovaries. After puncture the new follicular wave attained full numerical development within 3 d. Two-day intervals resulted in a lower number of follicles than the 3-d interval (11.0 -/+ 3.8 vs 15.4 -/+ 6.1; P < 0.05). In contrast 4- and 5-d intervals between puncture resulted in an increase in follicle size when compared with that of the 3-d interval. The mean+SD number of recruited follicles varied between animals ranging between 78 +/- 2.5 to 19.2 -/+ 6.0. The mean number of follicles recruited increased from the first month (March) to the third month (May) of sampling (11.8 +/- 4.7 vs 16.4 +/- 6.5; P < 0.01), and then decreased between the third (May) and the sixth (August) month of sampling (21.5 +/- 4.7 vs 16.8 +/- 5.0; P < 0.01). During the experiment, the number of recruited follicles varied cyclically, with waves having a length of 6 puncture session (PS) or 3 wk (i.e., the mean length the bovine estrous cycle). Follicular recruitment after repeated ovum pick-up showed a high repeatability (r = 0.576) A model was also developed showing good predictability of the potential of animals to recruit follicles on the basis of the first 4 to 6 PS. Our results showed that despite large variation in follicular recruitment between animals, the high repeatability and good predictability of follicle recruitment demonstrates the possibility of characterizing animals on their potential for follicle growth.

Journal Article↗

PubMed1995

Comparison of the efficacy of conventional slow freezing and rapid cryopreservation methods for bovine embryos.

Day 7 bovine morulae and early blastocysts were randomly assigned to one of four cryopreservation methods: (i) a modified conventional controlled slow freezing and stepwise dilution after thawing; and three methods which enable direct transfer of the embryo into the recipient upon thawing: (ii) conventional controlled slow freezing and a modification of a one-step procedure, (iii) vitrification with 6.5 M glycerol plus 6% BSA (w/v), and (iv) vitrification with 25% glycerol (v/v) and 25% propanediol (v/v). In a comparative in vitro study, the percentage of grade 1 and 2 embryos developing into expanded blastocysts in culture for cryopreservation methods 1-4 were, respectively, 53% (29/55), 33% (20/61), 44% (26/59), and 51% (17/33). Method 2 yielded a significantly lower survival rate than methods 1 (P < 0.1) and 4 (P < 0.05) and was excluded from a subsequent test of in vivo development. Pregnancy rates (Day 60) after transfer of embryos cryopreserved by methods 1, 3, and 4 were, respectively, 59% (20/34), 43% (17/40), and 24% (5/21). Method 4 yielded a significantly lower pregnancy rate than method 1 (P < 0.05). Method 3, however, did not yield a statistically different pregnancy rate (P > 0.1) when compared to method 1. Method 3 has considerable promise in providing a successful method for the cryopreservation of bovine embryos that (i) reduces the time required for equilibration and cooling, (ii) provides for simple and rapid one-step dilution of cryoprotectant after thawing, and (iii) enables more embryos to be thawed and transferred per unit time.

Animals↗

PubMed1995

Culture of bovine embryos to the blastocyst stage using Buffalo rat liver (BRL) cells.

A comparison was made between the development of in vitro matured and fertilized bovine oocytes in co-culture with bovine oviduct epithelial (BOE) cells or with Buffalo rat liver (BRL) cells. Both cell types supported development from the 1-cell to the blastocyst stage with equal efficiencies (4.4% for BRL cells, 4.0% for BOE cells). Medium conditioned by either cell type supported development to the blastocyst stage as efficiently as co-cultures (6.4 and 7.3% blastocysts for BOE and BRL conditioned medium, respectively). A higher percentage of blastocyst development was found when embryos were cultured closely apposed in small drops of BRL-conditioned medium compared with larger volumes (20.5 versus 7.0%). The ability of BRL-conditioned medium to support embryonic development was dependent on the duration of the conditioning period (optimum 24 to 48 h), and was not lost when the medium was stored at -20 degrees C for extended periods. The effects were independent of the conditions used to promote maturation in vitro and the procedure for fertilization. With 2 different methods to produce embryos in culture, both the BRL cell co-culture and BRL-conditioned medium in microdrops supported embryo development to the blastocyst stage. The use of the BRL cell line reduces the variability associated with primary BOE cell cultures.

Journal Article↗

PubMed1994

[Possibilities of ovum pickup in cattle].

Cattle breeding has a new reproduction technique in addition to artificial insemination and embryo transfer. It is the collection of ova from living animal by means of ultrasound guided follicle aspiration (ovum pick-up), followed by embryo-production in vitro. Follicles larger than 2 mm were punctured and the ova were collected twice weekly during 3 months. In total 1677 ova were collected from ten cows; 1342 (80%) were used for in vitro maturation, fertilization, and embryoculture. All ova were fertilized with semen from one bull, and 218 transferable embryos were produced. Calculated on a year basis, this would amount to 87 embryos per animal, with an intra-animal variation between 28 and 132. This new technique may replace MOET (Multiple Ovulation and Embryo Transfer; yearly average of 25 transferable embryos per animal), if the embryo-production via OPU can be performed with semen from any selected bull.

Animals↗

PubMed1994

Potential use of ovum pick-up for embryo production and breeding in cattle.

The efficacy of transvaginal ultrasound-guided puncturing of ovarian follicles for collecting immature oocytes in cattle was studied. Three experiments were conducted to examine the effects of puncturing on follicle recruitment and on the number of oocytes collected. Puncture sessions were executed twice weekly at regular intervals of 3 and 4 d respectively. The oocytes were matured, fertilized and allowed to develop in vitro and the number of transferable embryos was recorded. The health of the cows was checked daily. In Experiment 1, dairy cows (n=10) were punctured over a period of 5 mo, and the collected oocytes were fertilized with the semen of 1 bull. In Experiment 2, oocytes were collected from one 12 year old high pedigree dairy cow and an one month pregnant cow were punctured. The oocytes of the old cow were fertilized with semen of 8 different bulls. In Experiment 3, beef cows (n=6) were punctured over a 2 mo period and the semen of 2 different bulls of the same breed was used to fertilize the oocytes from 3 of these cows. In Experiment 1, 14.5 +/- 0.4 (mean +/- SEM) follicles were punctured per session, and 8.0 +/- 0.3 (mean +/- SEM) oocytes were recovered. A mean of 16% of the oocytes developed into transferable embryos with a pregnancy rate of 40%. The results did not differ between the months of the experiments, indicating that the transvaginal puncturing method can be used successfully over a 5 mo period. No detrimental effects were observed after clinical and post mortem examinations, nor did breed, age or reproductive status appear to affect the results. However, large differences were observed between individual cows and between cow/bull combinations.

Journal Article↗