Biomedical subjects

R H Foote

Publications and source records attributed to R H Foote.

At least 109 records · Page 6Linked to original sources

PubMed1990

Effects of superovulation, embryo recovery, culture system and embryo transfer on development of rabbit embryos in vivo and in vitro.

Uninterrupted development of rabbit embryos in vivo was studied in 7 superovulated and 7 normally ovulating (GnRH-treated) does, while another 7 does were superovulated and 1-cell embryos were collected from them at 19 h after LH to compare development in vivo and in vitro. Embryos from the last group were either cultured in the presence or absence of rabbit oviduct epithelial cells for 65 h in Medium 199, or were immediately transferred to recipients. At 84 h after LH or GnRH, blastomere number, embryo volume and stage of development were assessed for all embryos. Intrazonal embryo volumes were significantly reduced in embryos recovered from superovulated donors. Superovulation also had a negative effect on embryo cell numbers. However, this reduction was more severe in embryos remaining in vivo in superovulated donors until 84 h after LH than it was in embryos transferred to nonsuperovulated recipients at the 1-cell stage (19 h after LH). The embryo recovery procedure apparently caused little harm to the embryos, except that the mucin layer on flushed and immediately transferred embryos was significantly thinner than that of embryos residing continuously in vivo. Co-culture with rabbit oviduct epithelial cells resulted in improved development in vitro, but this development was still significantly retarded compared with embryos developing in vivo.

Animals↗

PubMed1990

Comparison of persistence of seven bovine viruses on bovine embryos following in vitro exposure.

The ability of seven cytopathic strains of bovine viruses to adhere to the zona pellucida of six-to-eight day-old bovine embryos were compared. Embryos were exposed to virus by placing them either in virus suspensions or by culturing them on infected bovine turbinate cultures for 18-24 h. After exposure to bovine virus diarrhea virus (BVDV), infectious bovine rhinotracheitis virus (IBV), bluetongue virus (BTV), pseudorabies virus (PRV), vesicular stomatitis virus (VSV), parainfluenza 3 virus (PI3), or bovine enterovirus virus (BEV), the embryos were tested for virus by culture in bovine turbinate cells and by morphological examination using electron microscopy (EM). A special technique to minimize loss of embryos processed for EM was developed. More embryos had viral particles on the surface of the zona pellucida after exposure to 18-24 hour infected cell cultures than did embryos exposed to viral culture suspensions. The most dramatic finding was that BTV adhered in large numbers to the surface of the zona pellucida of exposed embryos. IBRV, PRV, and VSV comprised an intermediate group, with virions occasionally detected on the surface of exposed embryos after 5 washes. Therefore, extensive washing is required. The PI3 and BEV were easily removed from embryo-exposed virus by washing. BVD was difficult to identify morphologically, making assessment by EM unreliable. There was no evidence that any one of the seven viruses penetrated the intact zona pellucida. Using a micromanipulator, 42 embryos were also directly inoculated through the zona pellucida with +/- 50 picoliters of virus inoculum or medium.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1990

Method for obtaining bovine zygotes produced in vivo.

A superovulatory and surgical protocol was developed for recovery of bovine zygotes. Holstein cows and heifers were given follicle-stimulating hormone and cloprostenol to induce superovulation. Surgical cannulation and lavage of the uterine tube was performed 40 to 48 hours after the start of standing estrus. In general, cows had more corpora hemorrhagica than did heifers, but a higher percentage (P less than 0.05) of ova recovered from cows were infertile. Several heifers were subjected to the procedure twice, and embryo recovery rates were equivalent both times.

Age Factors↗

PubMed1990

Experimental transmission of bovine viral diseases by insemination with contaminated semen or during embryo transfer.

Three experimental approaches were used to study transmission of blue tongue (BT), infectious bovine rhinotracheitis (IBR) and bovine virus diarrhoea (BVD) viruses. These were insemination with contaminated semen, experimental infection of embryo donor cows, or transfer of embryos experimentally exposed to virus in vitro to normal recipients. Parameters assessed included number and quality of embryos produced, virus detection (isolation and electron microscopy), serology and histopathology. All superovulated sesceptible cows inseminated with semen containing blue tongue virus (BTV) (n = 2) or infectious bovine rhinotracheitis virus (IBRV) (n = 2) became infected. One cow inseminated with semen containing BTV produced seven virus-free seven-day-old embryos; the second cow failed to produce any embryos. One of two cows inseminated with semen containing IBRV produced two underdeveloped, virus-free embryos while no embryos were produced by the second cow. One of two cows inseminated with semen containing bovine viral diarrhoea virus (BVDV) became infected. Two poorly developed, virus-free seven-day-old embryos were recovered from one of these cows. Superovulated susceptible cows inoculated either intramuscularly with BTV (n = 3) or intranasally with IBR virus (n = 2) became infected. Virus was isolated from some tissues of two BTV-infected cows, neither of which produced embryos. A third BTV-infected cow produced two virus-free embryos collected at necropsy five days after inoculation. One of two cows experimentally infected with IBR virus, produced three embryos but virus was not detected either by electron microscopy (1 embryo) or in cell culture by cytopathic alterations (1 embryo).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1990

[The effect of gonadotropin releasing hormone (GnRH) and analogs on the conception rate in cattle. A critical literature review].

The objective of this article was to review the current literature of the influence of gonadotropin releasing hormone (GnRH) and GnRH agonists on conception rate in dairy cattle. The application of GnRH or agonists at artificial insemination (first and subsequent) and between days 7 and 34 after parturition were considered. The variations between studies were discussed as well as different mechanisms concerning the influence of GnRH for establishing pregnancy. From a critical point of view the routine use of GnRH or agonists at the time of first or subsequent breeding or during the postpartum period cannot be supported.

Animals↗

PubMed1989

In vitro capacitation and fertilizing ability of ejaculated rabbit sperm treated with lysophosphatidylcholine.

Four experiments were replicated 1) to establish dose-response relationships between lysophosphatidylcholine (LPC), sperm motility, and the acrosome reaction (AR), 2) to evaluate the influence of rabbit serum (RS) on these endpoints, 3) to compare buck differences in induction of the AR, and 4) to examine fertilizing ability in vitro of sperm tested under the first three objectives. Semen was collected from Dutch-belted rabbits, washed once by centrifugation, resuspended, and preincubated for 2 or 4 hr in a chemically defined medium (DM), DM plus 20% RS, or BSA-free DM plus 20% RS at 37 degrees C. At the end of preincubation LPC was added to the preincubated sperm at concentrations of from 0 to 100 micrograms/ml. Sperm were examined .5-4 hr later for AR and sperm motility. For in vitro fertilization, sperm and ova were coincubated in DM up to 24 hr after insemination and in a more complex medium for another 24 hr. Addition of LPC to 4-hr-preincubated sperm was more effective for inducing the AR than addition to 2-hr-preincubated sperm. A significant increase (P less than .05) in the AR occurred in 15 and 30 min following exposure to 100 and 80 micrograms of LPC per ml, respectively, but the higher concentration of LPC decreased sperm motility. Addition of 20% RS to DM with or without BSA surprisingly inhibited the AR but maintained sperm motility, as expected. Bucks differed (P less than .05) in the initial percentage and the induced percentage of AR sperm. For the AR the optimal concentration of LPC per ml was 80 micrograms, but for in vitro fertilization 60 micrograms tended to be superior.

Animals↗

PubMed1989

A spectrophotometric procedure for the determination of objective measurements of equine spermatozoan motility.

A spectrophotometric procedure was developed and evaluated for the objective measurement of equine spermatozoan motility. A 100 mul sample of a sperm suspension, prepared by the removal of seminal plasma, was layered under a column of optically clear medium in a specially designed spectrophotometric cuvette maintained at 37 degrees C. Changes in light transmittance above the interface of the sperm suspension and medium were recorded on chart paper. As sperm cells swam into the medium, a decrease in light transmittance was recorded as a deflection on the chart paper. Chart recordings were analyzed for the height (cm) and time (min) to the peak deflection. To standardize the procedure, a fixed number of cells (1x10(9)) were used to prepare suspensions of 300x10(6) cells/ml. Coefficients of variation for mean values obtained under these conditions after the evaluation of five ejaculates from a given stallion were estimated at between 10 and 12%. Correlations between swim-up measurements and computer-assisted semen analysis demonstrated that the percentage of motile cells and mean velocity (mum/sec) of motile cells influenced swim-up measurements. Described here is a simple and inexpensive procedure to determine objective measurements of spermatozoan motility that may have application in semen evaluation and fertility testing in the stallion.

Journal Article↗

PubMed1989

Buserelin in a superovulatory regimen for Holstein cows. I. Pituitary and ovarian hormone response in an experimental herd.

Two experiments were conducted with frequent blood sampling in standard superovulatory regimens using follicle stimulating hormone (FSH) and prostaglandin F(2) alpha (PGF) to study the effects of the gonadotropin releasing hormone analog, Buserelin, on changes in FSH, luteinizing hormone (LH), progesterone (P(4)) and estradiol (E(2)). In Experiment I, Buserelin (20 mug) was administered to a total of 28 dry Holsteins. One group was treated with Buserelin 36 and 60 h after PGF administration, a second group was treated 60 h after PGF, and a third group served as the controls. In Experiment II, 30 dry Holsteins received Buserelin (10 mug). One group was treated 48 h after PGF, a second group at 54 h after PGF, a third group 24 h after estrus was first observed and a fourth group was a control. The general pattern of a decrease in P(4) following PGF, an increase in E(2), the onset of estrus, an LH peak, and finally, an increase in P(4) in superovulated cows was observed. Buserelin consistently produced a sharp LH peak at 36 h when given 36 h after PGF. At later intervals, it produced either a major or minor peak depending upon whether a spontaneous LH peak had already occurred. There was too much individual cow variation in the interval from PGF to a spontaneous LH peak to consistently induce a uniform LH peak, except when Buserelin was given 36 h after PGF, which may be early for normal oocyte maturation. There was no treatment effect on FSH, and embryo recovery rate was unaffected by treatment (P>0.05).

Journal Article↗

PubMed1989

Buserelin in a superovulatory regimen for Holstein cows: II. Yield and quality of embryos in commercial herds.

The gonadotropin releasing hormone analog, Buserelin, was tested in a superovulatory regimen in cows by administering 8 mug of it at the following times: Group I (12 cows), 48 h after the first prostaglandin F(2) alpha (PGF) injection: Group II (11 cows), 54 h after PGF: Group III (10 cows), 24 h after standing estrus was first observed; and Group IV (12 cows), served as superovulated controls. The cows were lactating Holsteins between 45 and 143 d post partum, with at least one estrus prior to superovulation. The number of embryos collected from Groups I, II, III and IV 7 d after estrus averaged 4.5, 8.1, 6.4 and 5.6, respectively (P>0.05). The fertilization rate in the three groups receiving Buserelin was 83 versus 76% for controls (P<0.10). Blood and milk samples taken just before starting follicle stimulating hormone treatment at the expected estrus and at the time of embryo recovery were tested for progesterone concentration, and results from a rapid ELISA test were useful in identifying cows that a) were unsuitable for superovulation, b) should have been in estrus but were not observed standing and c) produced few, if any, embryos.

Journal Article↗

PubMed1989

Optimal replication for histometric analyses of testicular function in rats or rabbits.

Quantitative evaluations of testicular histology can provide sensitive endpoints for determining toxicity of chemicals to the male reproductive system. But, the numbers of observations per testis or number of animals per treatment group often are selected by tradition or availability, rather than from a statistical basis. Therefore, we studied the number of observations per male (sampling intensity) and number of animals per treatment (replication) needed to detect treatment effects of given magnitude, with predictable error probabilities, using data from Sprague-Dawley rats and Dutch-belted rabbits that received 0.0, 0.94, 1.88, 3.75, 7.5, or 15.0 mg/kg body wt of 1,2-dibromo-3-chloropropane (DBCP). Data for one testis from 102 rats and 34 rabbits were available. For each testis, observations included measurement of the minor diameter of 50 seminiferous tubules, counts of the number of leptotene primary spermatocytes per 250 Sertoli cells, and counts of spherical spermatids within 20 seminiferous tubular cross sections. Tabular data are presented showing optimal numbers of observations per testis and animals per treatment group as a function of the difference to be detected and selected probabilities for Type I and II errors. In general, precise assessments required far fewer observations per testis than are used routinely. However, due to the inherent variability among animals, the number of animals required per treatment tended to be greater for experiments with the rabbit, and increased substantially for both species when detection of small differences had to be ensured. The data presented should enable investigators to design experiments of chosen sensitivity and precision while making cost-effective use of animals and labor.

Animals↗

PubMed1989

Fertility of fresh and frozen rabbit semen inseminated at different times is indicative of male differences in capacitation time.

Some reports indicate that sperm from different males differ in capacitation time, and other reports suggest that freezing sperm may affect their capacitation time. These two variables were specifically studied in rabbits in a fertility trial with 96 does inseminated with approximately 1.6 million motile fresh or frozen sperm from three different bucks at 15, 10, 5, and 0 h before expected ovulation. Fresh semen averaged 84% live (unstained) sperm and 88% had normal acrosomes; corresponding values for frozen sperm were 44% and 54%. On the basis of does that became pregnant, average litter size with fresh semen was 5.5 and with frozen semen was 4.8 (p greater than 0.05), but overall, does bred with frozen semen produced fewer young (p less than 0.05). On the basis of total does and total semen, average litter size from insemination at 15, 10, 5, and 0 h was 2.8, 4.2, 3.8, and 1.7, and average litter size for the three bucks was 4.0, 1.8, and 3.6. There was no interaction of type of semen (fresh or frozen) with the other variables in the model (p greater than 0.05). Bucks and time of insemination affected both the proportion of does that were pregnant and litter size (p less than 0.01). A major interaction between buck and time of insemination (p less than 0.01) was due apparently to both differential sperm survival and probable capacitation time among bucks. This major interaction should be considered in designing in vitro and in vivo fertility studies, and for selecting males for use in artificial insemination.

Acrosome↗

PubMed1989

Value of testicular and sperm profiles in optimizing reproductive success: lessons learned from selective breeding programs of domestic and laboratory animals.

The economic potential of the biotechnology of artificial insemination for genetic improvement and disease control was recognized in the United States 50 years ago. To fully exploit this technology in dairy cattle, researchers worked closely with industry to study testicular function, best methods for harvesting and evaluating semen, techniques for preserving and inseminating sperm and finally, methods for mass recording of fertility information. Methods of testicular and semen evaluation have been developed which are highly correlated with other reproductive characteristics, including fertility. This has resulted in an unprecedented opportunity to study experimentally imposed conditions and naturally occurring environmental conditions upon reproductive efficiency in the male. The selection and testing of sires has resulted practically in the elimination of genes which produce lethal or severely depressed performance. The use of one male for the controlled insemination of many females also has facilitated studies on reproductive problems in the female. While less selective breeding programs have been conducted with the rabbit relative to reproductive function, the rabbit has been used extensively as a model for large domestic animals and humans. The rabbit is the smallest laboratory animal which can be trained easily to ejaculate into an artificial vagina and use the semen for insemination. This permits longitudinal and correlative studies to be performed relating testis function and epididymal transport to sperm output, semen quality and fertility. Spermatogenesis of both bulls and rabbits is sensitive to toxicants. The effects can be monitored by testicular biopsy and cytological evaluation of the testis, by semen collection and semen evaluation, and by in vitro and in vivo fertility testing. Gross effects on the testis are detectable in the live animal by measuring testis size and by ultrasound profiles.

Animals↗

PubMed1988

Cholesterol efflux from bovine sperm. I. Induction of the acrosome reaction with lysophosphatidylcholine after reducing sperm cholesterol.

Methods were developed to quantitatively reduce the cholesterol (Chol)/phospholipid (PL) ratio of bovine sperm and to determine the effectiveness of this treatment in capacitating sperm. Washed sperm (2 x 10(8)) were incubated in 1.0 ml of modified Tyrode's solution (TS) containing unilamellar liposomes of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and [14C]-Chol (35:35:30 molar ratio, 300 nmol total PL). [3H]-triolein was included as a nonexchangeable marker. After 90 min at 39 degrees C, a 13% net exchange of [14C]-Chol from liposomes to sperm was observed (n = 4), and sperm motility was 80%. Sperm were then washed and 50 x 10(6) sperm were incubated as before with PC/PE liposomes containing no Chol. After 90 min, sperm were separated from liposomes by centrifugation. Measurement of [14C]-Chol in the liposomes (supernatant) and parallel gas chromatographic analysis of extracted, saponified liposomes (n = 4) indicated that 30% of sperm Chol was removed by this procedure. Chol efflux decreased percent motile sperm by less than 10% but reduced sperm velocity by more than 50%. Sperm incubated with no liposomes (control), with liposomes containing Chol (+Chol), and with Chol-free liposomes (-Chol) were washed and resuspended in TS with 0.2% BSA and 30 micrograms lysophosphatidylcholine (LPC)/mg bovine serum albumin (BSA). Percent sperm undergoing the acrosome reaction (AR) upon incubation with LPC-BSA was used as a measure of sperm capacitation. After 60 min of exposure to LPC-BSA at 39 degrees C, the mean (+/- SE) percent motile sperm for control, +Chol, and -Chol treatments was 57.0 +/- 4.9, 60.0 +/- 4.7, and 57.0 +/- 6.8, respectively. Corresponding values for percent AR were 14.0 +/- 3.4, 20.3 +/- 4.4, and 39.7 +/- 1.2. These results suggest that loss of Chol from bovine sperm may be an early step in sperm capacitation in this species.

Acrosome↗

PubMed1988

Cholesterol efflux from bovine sperm: II. Effect of reducing sperm cholesterol on penetration of zona-free hamster and in vitro matured bovine ova.

Several reports have indicated that sperm capacitation includes loss of membrane cholesterol (Chol) with a concomitant decrease in the Chol-to-phospholipid (PL) ratio. Methods were developed for quantifiable removal of bovine sperm Chol, which predisposed sperm to induction of the acrosome reaction upon addition of lysophosphatidylcholine (LPC). The objective of this study was to evaluate the effect of Chol removal from bovine sperm on penetration of zona-free hamster and intact bovine ova in vitro. Washed ejaculated bovine sperm were incubated (2 h, 39 degrees C) in a modified Tyrode's solution (TALP) containing 1) Chol-free liposomes (-Chol, 50 x 10(6) sperm and 600 nmol phospholipid/ml); 2) liposomes containing 30 mol% Chol (+Chol, 2 x 10(8) sperm and 300 nmol total lipid/ml); or 3) no liposomes (Control). We have previously shown that net Chol efflux from sperm is 31% of the total sperm Chol with -Chol liposomes and less than 1% with control media. Sperm were then washed twice and challenged with LPC bound to bovine serum albumin (BSA) using celite as a carrier. Treated sperm (25 x 10(6)) were incubated immediately with either zona-free hamster ova (HO) or in vitro matured bovine ova (BO) in 50-microliters droplets of TALP under medical fluid in an atmosphere of 5% CO2 in air (3 h, 39 degrees C). Ova were fixed in ethanol:acetic acid, stained with 1% orcein, and examined.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1988

Production of chimeric rabbits from morulae by a simple procedure.

Experiments were conducted to develop a simple and reliable technique to produce chimeric rabbits from morula stage embryos. In Experiments 1 and 2, an in-vitro test of viability was initially performed by culturing embryos to the blastocyst stage. Ninety-three percent of the "chimeric" embryos developed to the blastocyst stage compared to 94% for controls when embryos were manipulated soon after collection (Exp. 1). Eight-one percent chimeric embryos and 78% control embryos developed to blastocyst stage when embryos were held at room temperature for 4 hr (Exp. 2). In Experiment 3, enough morula-stage embryos were available from true breeding Dutch-belted and albino rabbits to form potentially 67 diverse "color" pairs. These micromanipulated pairs of morulae were successfully combined to produce 64 chimeric embryos (96%, 64/67). They were transferred to the uteri of seven recipient does and three became pregnant producing 13 young. Four of the young exhibited substantial overt chimerism (31%) and one more was a possible chimera.

Animals↗

PubMed1988

An enzyme-linked immunoassay of milk progesterone as a diagnostic aid in embryo transfer programs.

The objective of this study was to use an enzyme-linked immunosorbent assay to measure progesterone (P(4)) in donor cows as an indicator of reproductive normality and response to superovulatory treatment. Forty-eight Holstein cows were superovulated, and blood and milk samples were collected four times: 1) at initial follicle stimulating hormone treatment, 2) just prior to administration of prostaglandin F(2)-alpha, 3) at estrus and 4) at the time of embryo recovery, 6 to 7 d after estrus. Cows were inseminated twice during estrus. The P(4) profiles (ng/ml) obtained at the four collections, were, respectively, as follows: blood plasma, 2.4, 2.7, 0.2 and 11.2; defatted milk analyzed by radioimmunoassay, 3.7, 3.8, 0.5 and 14.6; preserved whole milk analyzed by ELISA, 6.3, 7.8, 0.7 and 19.1; frozen whole milk analyzed by ELISA, 9.8, 10.0, 0.8 and 21.1. All four assays resulted in the expected pattern of P(4) values for most cows at the four stages sampled. Approximately 10% of the cows had P(4) concentrations outside of the expected range of values. A majority of these cows were aberrant, based upon behavior and palpation of the reproductive organs. The rapidly available P(4) results from the ELISA were useful in making appropriate decisions regarding management of donor cows.

Journal Article↗

PubMed1988

Objective analysis of stallion sperm motility.

An image-analysis system utilizing a microcomputer and CellSoft computer-assisted semen analysis software package was evaluated to assess stallion sperm motility characteristics. Analyses were performed at 37 degrees C on a 6 microl drop of diluted semen placed on a glass slide and covered with an 18 mm(2) coverslip. Four groups of 25 cells each per slide, four slides per ejaculate and four ejaculates from each of three stallions were analyzed in a nested model. The percentage of motile sperm cells, mean velocity (microm/sec), mean linearity, and mean angular head displacement (microm) were measured. Statistical analysis of variance components showed that within ejaculates, more variation was accounted for in the differences among groups of 25 cells than among slides. Predicted standard deviations calculated for combinations of slides and groups of cells showed that a combination of two slides from which a total of 400 cells were analyzed resulted in a mean intra-assay coefficient of variation (CV) of 5.7% for the four measured variables. The following are individual coefficients of variation: percentage of motile cells (7.8%), mean velocity (6.4%), mean linearity (1.9%) and mean angular head displacement (6.6%). When ejaculate differences were included in the model and predicted standard deviations were calculated for a single ejaculate, the mean inter-assay CV was 9.2%. Mean velocity (6.4%) and mean linearity (4.7%) were more repeatable among ejaculates than either the percentage of motile sperm (14.4%) or angular head displacement (11.2%). It was concluded that this system is precise enough to determine differences in motility characteristics of stallion semen samples.

Journal Article↗

PubMed1988

Factors limiting reproductive efficiency in selected laboratory animals.

The mouse, rat, hamster, and rabbit have had remarkable success in surviving in the wild, which attests to their high reproductive capability. In fact, Parkes refers to mice conceiving at each postpartum estrus having the potential for 13 litters per year. Paufler et al. used GnRH to repeatedly cause ovulation in 27 rabbits a few days after parturition, resulting in an average pregnancy rate of 71.5%, 7.0 young born, 5.8 young weaned, and 50 young per doe per year. All four species produce an excess of spermatozoa relative to the requirements for fertilization. The rabbit is suitable for semen collection, artificial insemination, and most of the techniques one might wish to model for reproductive studies in both males and females. A major limitation to reproduction in these species is that reproductive capacity fails long before the finite oocyte population formed prenatally is depleted. The uterus of the aged female appears to be the major cause of reproductive failure, as fertilized eggs replaced in such a uterus usually soon deteriorate. With in vitro techniques, much is still to be learned about harvesting, maturing and fertilizing oocytes and identifying those most likely to result in formation of a healthy neonate.

Animals↗