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Biomedical subjects

D C Kraemer

Publications and source records attributed to D C Kraemer.

At least 19 recordsLinked to original sources

Effects of stage of oestrous cycle and progesterone supplementation during culture on maturation of canine oocytes in vitro.

The aim of this study was to evaluate the effect of progesterone supplementation and stage of oestrous cycle on in vitro maturation (IVM) of canine oocytes. Oocytes were cultured in medium supplemented with 0, 2000, 4000 or 8000 ng progesterone ml(-1) (Expt 1; n=274 oocytes) or 0, 20, 200 or 2000 ng progesterone ml(-1) (Expt 2; n=789 oocytes). In Expt 3, oocytes (n=1202) were cultured in a bi-phasic system of meiotic arrest followed by IVM, both in the presence of 0, 20, 200 or 2000 ng progesterone ml(-1). Rates of meiotic resumption for Expt 1 ranged from 40.0% to 58.5%; there were no significant differences among groups. In Expt 2, rate of meiotic resumption was significantly lower in the 2000 ng progesterone ml(-1) treatment (35.5%) compared with the 200 ng progesterone ml(-1) treatment (54.0%; P<0.05). There were no significant differences in rates of maturation to metaphase II among treatments in Expt 1 (1.8-8.6%) or Expt 2 (8.4-14.7%); however, oocytes collected from ovaries of bitches in oestrus and dioestrus had higher rates of maturation to metaphase II than did oocytes from bitches at pro-oestrus or anoestrus (P<0.01). In Expt 3, no differences were observed in rates of maturation among treatment groups. Rates of maturation to metaphase II of oocytes from bitches in dioestrus were significantly higher than those from bitches in pro-oestrus (P<0.01). These results indicate that supplementation of culture medium with progesterone either during maturation or during meiotic arrest before maturation does not increase the rate of IVM of canine oocytes. However, stage of oestrous cycle is a key factor in the selection criteria for meiotically competent canine oocytes for use in in vitro experiments.

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Effects of oocyte culture density on meiotic competence of canine oocytes.

This study was conducted to determine a suitable ratio of oocytes to medium for in vitro maturation (IVM) of cumulus-oocyte complexes (COCs) collected from bitches at anoestrus and dioestrus and to examine the meiotic competence of COCs cultured singly or in different group sizes. In the first experiment, different numbers of COCs (5, 10, 15 and 20 per drop) were cultured for 72 h in 100 microl drops of maturation medium. The meiotic competence of oocytes from ovaries at anoestrus was affected by the number of COCs incubated, whereas at dioestrus, the incubation number of COCs had no effect. In the second experiment, COCs were cultured singly or in different group sizes for 72 h by suitable oocyte density according to the reproductive cycle of the donor. In the anoestrous group, 1, 5 and 10 COCs were cultured in 10, 50 and 100 microl drops of the medium (10 microl per COC), respectively. In the dioestrous group, 1, 5 and 15 COCs were cultured in 7, 35 and 105 microl drops of the medium (7 microl per COC), respectively. There were no differences in the proportions of oocytes reaching metaphase II among the different group sizes in each stage of the reproductive cycle of the donor. The results indicate that the influence of oocyte density on the meiotic competence of oocytes differs according to the stage of the reproductive cycle of the donor. Moreover, the group sizes have no effect on the meiotic competence of oocytes cultured at suitable oocyte density according to the reproductive cycle of the donor.

Anestrus↗

Potential for cloning dogs.

The aim of this study was to determine whether nuclear transplantation could be used to clone a dog using donor nucleus cells collected from an adult female. Fibroblasts obtained from skin biopsies were fused with enucleated bovine or canine oocytes. The resulting cloned embryos were cultured in vitro to monitor embryonic development. A proportion of the resulting embryos was transferred into surrogate bitches for development to term. When canine oocytes were used as recipient ova for canine fibroblasts, 23% of the resulting embryos cleaved at least once after culture in vitro. Five cloned embryos were transferred into three synchronized recipient bitches, but no pregnancies resulted. When bovine oocytes were used as recipinets for canine fibroblasts, 38% cleaved to the two- to four-cell stage and 43% cleaved to the eight- to 16-cell stage. Forty-seven of these embryos were transferred into four recipient females, resulting in a single conceptus that ceased development at about day 20 of gestation. The desire for cloned dogs is considerable and will undoubtedly incite the development of successful methods for cloning companion animals. However, significant investment into additional research is required, especially in the areas of in vitro maturation of oocytes and control of the oestrous cycle of bitches.

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Genetic reprogramming of lactate dehydrogenase, citrate synthase, and phosphofructokinase mRNA in bovine nuclear transfer embryos produced using bovine fibroblast cell nuclei.

Adult animal cloning has progressed to allow the production of offspring cloned from adult cells, however many cloned calves die prenatally or shortly after birth. This study examined the expression of three important metabolic enzymes, lactate dehydrogenase (LDH), citrate synthase, and phosphofructokinase (PFK), to determine if their detection in nuclear transfer (NT) embryos mimics that determined for in vitro produced embryos. A day 40 nuclear transfer produced fetus derived from an adult cell line was collected and fetal fibroblast cultures were established and maintained. Reconstructed NT embryos were then produced from this cell line, and RT-PCR was used to evaluate mRNA reprogramming. All three mRNAs encoding these enzymes were detected in the regenerated fetal fibroblast cell line. Detection patterns were first determined for IVF produced embryos (1-cell, 2-cell, 6-8 cell, morula, and blastocyst stages) to compare with their detection in NT embryos. PFK has three subunits: PFK-L, PFK-M, and PFK-P. PFK-L and PFK-P were not detected in bovine oocytes. PFK subunits were not detected in 6-8 cell embryos but were detected in blastocysts. Results from NT embryo RT-PCR demonstrated that PFK was not detected in 8-cell NT embryos but was detected in NT blastocysts indicating that proper nuclear reprogramming had occurred. Citrate synthase was detected in oocytes and throughout development to the blastocyst stage in both bovine IVF and NT embryos. LDH-A and LDH-B were detected in bovine oocytes and in all stages of IVF and NT embryos examined up to the blastocyst stage. A third subunit, LDH-C was not detected at the blastocyst stage in IVF or NT embryos but was detected in all earlier stages and in mature oocytes. In addition, LDH-C mRNA was detected in gonad isolated from the NT and an in vivo produced control fetus. These results indicate that the three metabolic enzymes maintain normal expression patterns and therefore must be properly reprogrammed following nuclear transfer.

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A simple salt solution medium supplemented with yolk plasma and lactate (YPLM) supports development of preimplantation bovine embryos in vitro.

This study investigated the embryotropic potential(s) of egg yolk (EY) or its fractions, yolk plasma (YP) and yolk granules (YG), in culturing bovine embryos in vitro and substituting for protein (FCS, BSA, and BME-E and MEM-NE amino acids) and energy (glutamine, pyruvate and L-lactate) supplements commonly added to culture medium. In the first set of experiments (Experiment 1, 2 and 3) CR1aa with buffalo rat liver (BRL) cells were used as a co-culture system. The addition of 2.5% or 5% EY significantly increased (P<0.05) blastocyst percent over the BRL control (48.3% and 52.4% vs. 32.5%, respectively). The addition of 5% EY in the absence of FCS and BSA resulted in percent development to blastocysts and hatched blastocysts similar (P>0.05) to those of the BRL control (37.6% and 57.4% vs. 51.5%, 22.7% and 39.5%, respectively). The supplementation of the BRL control with 5% YP compared to that of EY resulted in comparable (P>0.05) percentages of blastocysts and hatched blastocysts (39.0% and 51.6% vs. 40.0% and 58.3%, respectively). In the second set of experiments, the embryotropic potential of YP was examined using a cell-free culture system and a simple salt solution (SS) of NaCl, KCl and NaHCO3 as the base medium. The supplementation of an energy-supplemented cell-free simple salt solution (E-SS) with 5% YP in the absence of supplemental protein resulted in percent development into blastocysts and hatched blastocysts comparable (P>0.05) to those of a BRL control (39.2% and 15.8% vs. 37.1% and 22.2%, respectively). The addition of YP to the simple salt solution with hemicalcium L-lactate as the only supplemented energy ingredient resulted in percentages of blastocysts and hatched blastocysts similar (P>0.05) to those obtained by the supplementation of all energy sources (27.4% and 15.6% vs. 36.4% and 14.0%, respectively). Increasing hemicalcium L-lactate level from 5 to 10, 20 or 25 mM resulted in a significant decrease (P<0.05) in percent development into blastocysts (36.5% vs. 24.8%, 11.6% and 6.7%, respectively). In conclusion, YP, with the advantage of being clearer than EY, is capable of sustaining embryo development to the blastocyst stage in a simple salt solution of NaCl, KCl and NaHCO3 supplemented with hemicalcium L-lactate.

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Effect of low temperatures on in-vitro matured bovine oocytes.

This research concerned effects of cooling in vitro matured bovine oocytes on subsequent fertilization and development in vitro. Oocytes were maintained at 39 degrees C (control), 20 degrees C, 10 degrees C or 0 degree C for 5, 10, or 20 min, then fertilized and cultured in vitro for 7 d. The proportion of fertilized oocytes that cleaved and developed to the morula/blastocyst stage was compared between different treatments. Duration of exposure had no effect on the results. Fertilization rate was higher (P < 0.05) for oocytes maintained at 39 degrees C (73.2%) than for oocytes cooled at 20 degrees C (58.6%), 10 degrees C (47.3%), or 0 degree C (36.9%). Cleavage rates were 58.3, 45.3, 15.7 and 7.0% for 39 degrees C, 20 degrees C, 10 degrees C and 0 degree C, respectively (P < 0.05). The lowest development rate to the blastocyst stage was obtained with oocytes cooled to 10 degrees C (0.0%) or 0 degree C (0.9%), followed by 20 degrees C (7.1%) and 39 degrees C (16.5%; P < 0.05). In a second experiment, the zona pellucida was removed after cooling but prior to fertilization (zona-free) from a portion of the in vitro- matured bovine oocytes in each treatment. When sperm penetration rates of zona-free oocytes were compared (percentage of oocytes exhibiting > or = 2 pronuclei), there was no difference (P > 0.05) between oocytes cooled at 0 degree C (59.7%) or 10 degrees C (67.9%). However, penetration rates in these 2 groups were lower (P < 0.05) when compared to zona-free oocytes cooled at 20 degrees C (83.1%) or those maintained at 39 degrees C (83.1%). Zona-free oocytes had higher penetration rates (P < 0.05) when cooled at 0 degree C (59.7%) or 10 degrees C (67.9%) than zona-intact oocytes cooled at 0 degree C (37.3%) or 10 degrees C (47.2%). However, there was no difference in the penetration rate when zona-free and zona-intact oocytes were cooled at 20 degrees C or maintained at 39 degrees C. These data demonstrate that cooling in vitro-matured bovine oocytes decreases the percentage of oocytes that undergo fertilization and subsequently develop in vitro. Moreover, at least part of the decrease in fertilization following oocyte cooling is due to effects on the zona pellucida.

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Ultrasonographic evaluation of the effects of exogenous gonadotropins on follicular recruitment and ovulation induction in the Atlantic bottlenose dolphin (Tursiops truncatus).

Transabdominal ultrasonography and serum steroid concentrations were used to evaluate the effects of exogenous gonadotropin administration on ovarian activity of two anestrous bottlenose dolphins (Tursiops truncatus). The gonadotropin used for follicular recruitment was PG600, which has 400 IU equine chorionic gonadotropin (eCG) and 200 IU human chorionic gonadotropin (hCG) activity per 5 ml. Ovulation induction was attempted with hCG. PG600 was administered in two doses of 20 ml (1,600 IU eCG and 800 IU hCG) and 12.5 ml (1,000 IU eCG and 500 IU hCG), respectively, 48 hr apart on days 0 and 2. On day 6, 1,500 IU of hCG was administered. Progesterone and total immunoreactive estrogens were determined before and after the gonadotropin administration. Bilateral ovarian ultrasonographic exams were performed daily on days 4-9 and on day 22. Serum immunoreactive estrogen concentrations were greater than the pretreatment concentrations after day 4 for both dolphins and remained elevated for the rest of the study. Serum progesterone concentrations rose above 1 ng/ml 2 days after hCG treatment and remained elevated for the rest of the study. Small antral follicles (< 0.5 cm) were initially observed bilaterally in both dolphins on day 4. In both animals on day 9, there were > 12 follicles/ovary, ranging from 0.5 to 1.5 cm. By day 22, the multiple follicles ranged from 0.5 to 4.5 cm in diameter. No ultrasonographic evidence of luteal formation was observed. The results indicate that 1) transabdominal ultrasonography can be used to detect and follow follicle growth in bottlenose dolphins; 2) bottlenose dolphins are sensitive to exogenous gonadotropins (multiple follicular recruitment of follicles occurred); and 3) until further ultrasonographic studies can be conducted to evaluate the effects of titrated doses of exogenous gonadotropins, these protocols should be considered unsuitable for ovulation induction.

Anestrus↗

Plasma vitamin E and other analyte levels in Nepalese camp elephants (Elephas maximus).

Plasma concentrations of alpha-tocopherol (vitamin E) and other analytes in Asian elephants (Elephas maximus) in Nepal were determined during typical work camp management of the elephants. Elephants foraged for food for 4-6 hr each day under the control of mahouts and were also provided daily with cut forage and supplements of unhusked rice, cane molasses, and salt. Blood samples were taken monthly for 1 yr without chemical restraint from 26 female elephants in four camps. Elephants were 6-60+ yr of age. Mean (+/-SEM) alpha-tocopherol concentration was 0.77+/-0.047 microg/ml, with a range of 0.23-1.57 microg/ml. Subadults had lower concentrations than did older elephants, and there were significant differences in mean concentrations from different camps and in mean monthly concentrations. Plasma alpha-tocopherol concentration appears to vary widely between individuals, and a single value of <0.3 microg/ml is not sufficient to diagnose incipient vitamin E deficiency. Mean (+/-SEM) plasma retinol (vitamin A) concentration was 0.063+/-0.003 microg/ml with a range of 0.01-0.12 microg/ml. Subadults had higher concentrations than did older elephants, and mean retinal values differed significantly among camps. Beta-carotene was not found in plasma. Twenty-five other analytes determined or derived were generally similar to those reported in other Asian and African (Loxodonta africana) elephants. Estimates of nutrient intake, based upon diet composition, suggested that dietary concentrations of zinc and sodium may have been marginal, but the absence of signs of any nutrient deficiencies indicates that dietary husbandry in these elephant camps was generally satisfactory.

Aging↗

Effects of low temperatures on in vitro produced bovine zygotes.

The objective of this research was to investigate the effects of cooling on the development of bovine zygotes. One-cell bovine embryos were maintained at 39 degrees C (control), 20 degrees C, 10 degrees C, or 0 degree C for 5, 10, or 20 minutes, then cultured in vitro for 7 days and the proportion of embryos developing to the compact morula or blastocyst stage compared between different treatments. Duration of exposure time had no effect on development. Development rates to the compact morula or blastocyst stage were 3.9%, 11.4%, 17.4%, and 24.4% for zygotes maintained at 0 degree C, 10 degrees C, 20 degrees C, and 39 degrees C, respectively, with differences in embryo yield between every treatment (P < 0.05). In a second experiment, bovine pronuclei (karyoplasts) and cytoplasts were cooled at 0 degree C or maintained at 39 degrees C for 5 minutes. Pronuclear transplantation was then utilized to create 4 types of reconstructed embryos, those with: 1) non-cooled pronuclei and non-cooled cytoplasm, 2) non-cooled pronuclei and cooled cytoplasm, 3) cooled pronuclei and non-cooled cytoplasm, and 4) cooled pronuclei and cooled cytoplasm. The proportion of embryos developing to the blastocyst stage was highest when non-cooled pronuclei were transferred into non-cooled cytoplasm (18.9%), and similar to that of non-cooled, non-manipulated control zygotes (13.2%, P > 0.05). No embryos developed to the blastocyst stage when pronuclei (cooled or non-cooled) were transferred into cooled cytoplasm. However, zygotes with cooled pronuclei transferred into non-cooled cytoplasm yielded 4.5% blastocysts (P < 0.05). More embryos developed to the compact morula or blastocyst stage when non-cooled vs. cooled cytoplasm was utilized, regardless of whether the pronuclei were cooled (P < 0.05). These data demonstrate that pronuclei are more tolerant to low temperature exposure than is ovum cytoplasm.

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Analysis of urinary progesterone metabolites with behavioral correlation in Guenther's dik-dik (Madoqua guentheri).

Urine samples and behavioral data were collected from nine adult female Guenther's dik-dik (Madoqua guentheri) from September 1989-November 1992. The durations of predefined individual behaviors were recorded during focal sampling periods and ad libitum. Immunoreactive pregnanediol-3-glucuronide (PdG) concentration in dik-dik urine was determined by radioimmunoassay. The immunoreactive PdG radioimmunoassay was validated for use with dik-dik by determining parallelism [F(alpha=0.05,1,4) = 0.04) between a serially diluted PdG standard and serially diluted dik-dik urine. Behavioral estrus was manifested by lordosis. Eighty-nine percent of the estrous behavior occurred during or within 2 days of the interluteal phase period, as delineated by immunoreactive PdG concentrations; thus, a high degree of correlation between hormone concentrations and behavioral data was observed. The mean (+/-SD) luteal phase, interluteal phase, and estrous cycle lengths were 14.4+/-5.5 days (range, 6-29 days; n = 50), 6.6+/-5.1 days (range, 2-33 days; n = 50), and 20.2+/-6.6 days (range, 10-43 days; n = 48), respectively. During the collection period, one animal conceived, with a gestation period of 170 days. Estrous cyclicity occurred throughout the year, with no evidence of seasonality. Cochromatography of pooled urine samples with radiolabeled PdG indicated that the major progesterone metabolite detected by the immunoreactive PdG antibody during luteal phase and pregnancy was not PdG. This is the first detailed description of female dik-dik reproductive endocrine activity.

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Interspecific transfer of IVM IVF-derived red sheep (Ovis orientalis gmelini ) embryos to domestic sheep (Ovis aries ).

Two embryo production methodologies were investigated to generate Red sheep embryos for use in an interspecific embryo transfer program. In Experiment 1, 4 multiparous female Red sheep (Ovis orientalis gmelini ) were implanted with CIDR type G devices for 11 d. Forty-eight hours prior to CIDR removal, a total of 22.5 mg bid of FSH-P was administered over a 3-d period. Laparoscopic embryo collection was performed 5 d post breeding, and embryos were transferred to domestic recipient ewes (Ovis aries and Ovis orientalis musimon ). In Experiment 2, 7 nulliparous female Red sheep were implanted with CIDR devices and injected with 200 IU of PMSG and 25 mg of FSH-P on the 8th day of implant insertion. At 60 to 70 h post PMSG/FSH-P treatment, follicular oocytes were aspirated laparoscopically. The recovered oocytes were matured in M199 (with fetal calf serum, FSH, LH, penicillin and streptomycin) at 39 degrees C in a humidified atmosphere containing 5% CO(2). At 24 h oocytes were fertilized with frozen-thawed semen at a concentration of 1.6 x 10(6) sperm/ml. The ova/embryos were placed in CR2 or BOEC culture medium at 20-22 h post IVF. Following 3 to 4 d in culture, embryos were transferred laparoscopically to the uterine horn of synchronized recipients. In Experiment 1, 4 embryos and 6 UFO were collected from 2 embryo donors, respectively. Two embryos were transferred with the aid of a laparoscope to each of 2 Rambouillet recipients, one of which gave birth to a healthy Red sheep lamb at 158 d of gestation. In Experiment2, a total of 62 oocytes was collected from 7 oocyte donors; 16 developed to the 16- to 32-cell stage and were transferred to 8 recipients. Three of these IVM-IVF embryos were transferred laparoscopically to 2 Mouflon recipients, resulting in no pregnancies. Thirteen IVM-IVF embryos were transferred to 6 Rambouillet recipients. Each of these gave birth to a single healthy Red sheep lamb. Gestation lengths of the 3 IVM-IVF lambs ranged from 152 to 162 d. This research demonstrates that when using compatible species IVM-IVF technology in conjunction with interspecific ET can lead to the production of live offspring and can be used to propagate exotic ovine species.

Journal Article↗

Nonsurgical recovery of degenerative ova from the uteri of mares.

Degenerated ova were recovered with and without viable embryos 6 days after ovulation in 30 of 210 collections from 24 of 66 mares. All ova were approximately 150 mum in diameter, with an intact zona pellucida and at various stages of cytoplasmic degeneration. Most of the ova were collapsed, although some had an oval appearance. Most of the ova were from maiden 2-year-old mares. Thirty-four of the ova were recovered from the first or second collections. Two ova were recovered from the third collection from two mares. Two degenerative ova per collection were recovered from five collections; three of these collections also contained viable embryos. Degenerated ova were recovered from three mares twice; but not from consecutive collection attempts. Recovered ova were fixed in 2% glutaraldehyde-PBS for scanning electron microscopy. These data indicate that not all unfertilized ova remain permanently in the oviduct; many traverse it and enter the uterus. Furthermore, these data also suggest that when degenerative ova pass into the uterus they either degenerate further and (or) move into the vagina. This is supported by the fact that not all ova can be accounted for when the uterus and (or) the oviducts are washed.

Journal Article↗

Viability and ultrastructure of equine embryos following culture in a static or dynamic system.

The viability and ultrastructure of equine embryos were assessed following culture in a static or perifusion system. The percentage change in diameter was greater (P less than 0.025) for embryos in the static treatment (71%) than in the perifusion treatment (33%). Fluorescein diacetate (FD) scores, the percentage of fluorescing cells (FC) and fluorescent intensity (FI), also were greater (P less than 0.05 and P less than 0.01) following static culture than for embryos cultured in the perifusion system. Four of 9 control embryos resulted in pregnancies but no embryos cultured in either system prior to transfer resulted in pregnancies. Three of 9 embryos in the static treatment group and 1 of 9 embryos in the perifusion group resulted in non-ecogenic vesicles. Ultrastructural evaluation agreed with FD scores and morphological assessment. The continued development of static cultured embryos without the presence of an inner cell mass (ICM) was supported by ultrasound data.

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Intrauterine insemination of farmed fallow deer (Dama dama) with frozen-thawed semen via laparoscopy.

Estrus and ovulation of mature fallow does (n=155) on two North American farms were synchronized by intravaginal silastic devices containing 0.3 g progesterone (CIDR-type G) for 14 d. Each of 151 does received laparoscopic intrauterine inseminations of either 50x10(6) (n=125) or 25x10(6) (n=26) frozen-thawed spermatozoa, 65 to 68 h after CIDR device withdrawal. Four does received intrauterine inseminations per vaginam of 50x10(6) spermatozoa 68 to 69 hours after CIDR device withdrawal. Semen from crossbred Dama dama damaxDama dama mesopotamica sires was collected in New Zealand by electroejaculation. The overall pregnancy rate to artificial insemination, as assessed by rectal ultrasonography at Day 45, was 67.7%. The pregnancy rates for does receiving laparoscopic inseminations were 58.2% (Texas; 50x10(6) spermatozoa; n=79 does); 80.8% (Texas; 25x10(6) spermatozoa; n=26 does) and 76.1% (New York; 50x10(6) spermatozoa; n=46 does). Three of the four does receiving intrauterine inseminations per vaginam became pregnant to the frozen-thawed semen.

Journal Article↗

Intraspecific blastocyst reconstitution in mice using giant trophectodermal vesicles produced by multiple embryo aggregation.

Giant trophectodermal (TE) vesicles, produced by aggregating multiple embryos, were evaluated as a means of enhancing the viability of reconstituted murine blastocysts. Previous studies have indicated that the development of chimeric conceptuses produced by multiple embryo aggregation is impaired, thus the initial objective here was to examine how this is affected by 1) differences in developmental rates of individual embryos comprising the aggregates, 2) lengths of in vitro culture, and 3) incompatibilities between maternal hosts and chimeric fetal allografts. The results indicate that the viability of aggregates was influenced by embryo genotype and length of in vitro culture. The average number of B6D2-F1 x BL/6 offspring resulting from aggregates cultured for 0.7 day was not significantly different (p greater than 0.05) from that obtained from either embryo singletons cultured for 0.7 and 1.7 days (2.9 +/- 0.3 and 3.9 +/- 0.4, respectively) or from nonmanipulated controls (3.2 +/- 0.2 and 4.3 +/- 0.3, respectively). Blastocyst reconstitution studies were then conducted using giant TE vesicles from B6D2-F1 x BL/6 mice and inner cell masses from C3H mice. The average number of C3H offspring produced (2.7 +/- 0.1) was not significantly different (p greater than 0.05) from that of B6D2-F1 x BL/6 embryo aggregates (3.2 +/- 0.6) or of nonmanipulated controls (4.3 +/- 0.3). The results demonstrate that giant TE vesicles can be effectively used for intraspecific blastocyst reconstitution in mice.

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Reproductive anatomy, manipulation of ovarian activity and non-surgical embryo recovery in suni (Neotragus moschatus zuluensis).

Marked disparity in the uterine horn dimensions and relative degrees of caruncle development in suni suggested that exclusive or predominant dextral implantation occurs in association with bilateral ovulatory activity. Daily urinary measurements of pregnanediol-3 alpha-glucuronide revealed an oestrous cycle of approximately 21 days in length. Ovarian activity was controlled for synchronization of oestrus by using progestagen-impregnated intravaginal sponges and multiple ovulations were induced by using exogenous gonadotrophin therapy. An effective transcervical uterine catheterization technique was developed for the non-surgical collection of embryos. The efficiency of embryo recovery performed 5 days after sponge removal was 50.0%.

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