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Biomedical subjects

R H Foote

Publications and source records attributed to R H Foote.

At least 73 records · Page 4Linked to original sources
PubMed1993

Factors affecting preservation and fertility of bull sperm: a brief review.

This paper is a brief review of the factors that determine the number of sperm required for insemination to obtain high fertility and ways that sperm viability might be prolonged. Damage to sperm during freezing results in a requirement, after thawing, of about 6 x 10(6) motile sperm (> 10 x 10(6) total) per insemination to achieve near-maximal fertility, whereas 2.5 x 10(6) motile fresh sperm result in high nonreturn rates. Multiple inseminations to bracket the time of ovulation are usually not economical except in superovulated cows. Earlier unpublished work on sperm packaging for slow release in the cow and methods for stabilizing membranes to increase sperm survival time in the cow are discussed. Current studies are directed towards reducing catabolic metabolism of sperm and studying membrane changes during freezing and thawing and during incubation with bovine oviduct epithelial cells. Studies with bull sperm indicate that the choline and ethanolamine phosphoglyceride components of their membranes represent an unstable configuration. Exposure of sperm to liposomes with the sterol cholesterol can alter the phospholipid bilayer and increase capacitation time. Similar approaches may produce sperm with a longer fertilizing life following insemination. New procedures in vitro permit low cost modelling of fertilization, which will facilitate research by reducing the cost of studies in vivo.

Animals↗
PubMed1993

Development of rabbit zygotes cultured in protein-free medium with catalase, taurine, or superoxide dismutase.

The present study was designed to examine the influence of several potential antioxidants in a synthetic medium completely devoid of macromolecules. Antioxidants may be beneficial as additives to a synthetic medium because they could serve as scavengers of toxic free radicals in media lacking serum, serum albumin, or other macromolecules that may serve as scavengers in more complex media. Rabbit zygotes were cultured for 72 h at 39 degrees C in media containing varying concentrations of the anti-oxidants in a gas phase consisting of 5% CO2:95% humidified air. At 72 h embryo development was recorded, and embryos were fixed and stained with Hoechst 33342 DNA stain to facilitate counting of cells. In experiment 1, concentrations of catalase ranging from 250 to 1000 IU did not affect the proportion of zygotes developing into blastocysts or the cell number (p > 0.05). In experiment 2, 2.5-10 mM taurine increased the proportion of zygotes developing into expanding blastocysts (70-78% vs. 40% for controls) and increased cell number from 100 to 154-159 (p < 0.05). In experiments 3 and 4, superoxide dismutase was tested over a range of 100-2400 IU/ml. The response reached a plateau at 600 IU/ml, with positive responses in rate of embryo development and growth similar to that found with taurine in experiment 2. These studies indicate that culture of zygotes into blastocysts in a macromolecule-free medium with 5% CO2:95% air is substantially improved when agents that could serve as antioxidants are included. Other mechanisms of action are not excluded by these studies.

Animals↗
PubMed1993

Culture of rabbit zygotes into blastocysts in protein-free medium with one to twenty per cent oxygen.

Embryos were collected from superovulated Dutch rabbits 19 h after injection of LH and insemination. The embryos were at the one-cell stage at that time and those judged to be normal by the absence of granular cytoplasm and regular shape were distributed randomly within donors into culture dishes containing 500 microliters of a macromolecule-free medium consisting of RPMI-1640 and low glucose Dulbecco's modified Eagle's medium, 1:1, without a cover of oil. In Expt 1, O2 concentrations of 5, 10 and 15%, with 5% CO2 plus 90, 85 and 80% N2, respectively, were tested. In Expt 2, O2 concentrations of 1, 5 and 20% were combined with 5% CO2 and the remaining gas was N2. After 84 h in culture at 39 degrees C, embryos were examined for stage of development and stained with Hoechst 33342 so that the number of cells could be counted. In Expt 1, the proportion of embryos reaching the hatching blastocyst stage after 84 h in culture in 5, 10 and 15% O2 was 48, 38 and 21%, (P < 0.01) and the cell number per embryo averaged 258, 226 and 188, respectively (P < 0.01). In Expt 2, the proportion of hatching embryos after 84 h in culture in 1, 5 and 20% O2 was 67, 72 and 29% (P < 0.01), respectively. Cell numbers in the 1 and 5% O2 concentrations were higher than in the 20% O2 concentration (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Survival of bull spermatozoa seeded and frozen at different rates in egg yolk-tris and whole milk extenders.

Six factorially arranged experiments were designed to study effects of seeding, freezing, and thawing rates in whole milk and egg yolk-Tris extenders commonly used for commercial cryopreservation of bull sperm. In these extenders, semen normally is supercooled to -13 or -14 degrees C unless the sperm are seeded. When sperm were supercooled or seeded, either mechanically or with immobilized silver iodide, and frozen to -196 degrees C, the postthaw percentages of motile sperm were 59, 57, and 64%, respectively. Freezing rates of -15, -25, and -35 degrees C/min gave similar sperm survival rates and were superior to -5 degrees C/min. For milk, the critical freezing temperature extended to -75 degrees C before transfer to liquid nitrogen gave good results. For egg yolk-Tris extender, transfer to liquid nitrogen was less critical once -50 degrees C had been attained. Thawing of sperm in water baths at 25 and 45 degrees C gave similar results, and both temperatures were superior to 5 degrees C. The postthaw percentage of motile sperm in egg yolk-Tris was equal or superior to that of sperm frozen in milk. A freezing rate of -15 degrees C/min to -100 degrees C and thawing at 25 degrees C consistently gave good results.

Animals↗
PubMed1993

Fertility of bull spermatozoa frozen in whole milk extender with trehalose, taurine, or blood serum.

Fertility of bull semen processed in heated whole milk-glycerol control semen extender with various additives was compared in six field trials. The additive in field trials 1 and 2 was 25.6 g of trehalose/L of the glycerol fraction of whole milk. Whole milk was heated to 95 degrees C for 10 min, cooled, and filtered 1 d before use (trial 1) or 3 d before use (trial 2). In field trial 3, 3.0 g/L of taurine were added to the glycerol fraction of whole milk. In field trial 4, specially prepared bovine serum (15% vol/vol) was included in the glycerol fraction of whole milk. Field trials 5 and 6 were larger fertility studies with trehalose in extenders prepared the day before use (trial 5) and 1 and 3 d before use (trial 6). Control and treated semen were coded and distributed randomly over a large group of professional inseminators. The 59-d nonreturn rates for control and treated semen, respectively, were as follows: trial 1, 74.1 and 73.7%; trial 2, 71.3 and 73.1%; trial 3, 74.9 and 70.9%; and trial 4, 75.1 and 71.6%. No significant differences resulted in trials 1 to 3, but bovine serum decreased the non-return rate in trial 4. Trials 5 and 6 resulted in nonsignificant improvement in fertility with added trehalose. Thus, these additives, useful as cryopreservatives or membrane protectors in other systems, did not enhance the fertility of sperm frozen in whole milk.

Animals↗
PubMed1992

Comparison of spermatozoal movement and semen characteristics with fertility in stallions: 64 cases (1987-1988).

Information pertaining to evaluation of single ejaculates of semen and records for 2 consecutive breeding seasons were obtained. In all, data for 99 individual breeding seasons (n = 43 Standardbreds and 56 Thoroughbreds) were evaluated. Included in each semen evaluation was examination of semen characteristics and computer-aided analysis of spermatozoal movement characteristics. On the basis of the analysis of breeding records for 4,175 mares (7,017 estrous cycles), a per-estrous cycle fertility rate was calculated from data for 96 of the breeding seasons. Stallions with lower fertility than the mean overall season fertility had significantly (P less than 0.01) lower mean values for subjective appraisal of the percentage of motile and progressively motile spermatozoa and for percentage of morphologically normal spermatozoa. Lower mean values were obtained for computer-aided movement analysis of the percentage of motile and progressively motile spermatozoa, and for mean velocity of motile spermatozoa. Semen characteristics, including spermatozoal movement characteristics, and fertility were significantly (P less than 0.05) correlated for Thoroughbred and Standardbred stallions when analyzed individually and when data for both breeds were combined. Characteristics most highly correlated (P less than 0.01) with fertility data for both breeds combined were: subjective appraisal of the percentage of motile (r = 0.40) and progressively motile (r = 0.46) spermatozoa; percentage of morphologically normal spermatozoa (r = 0.36); and computer-aided analysis of percentage of motile spermatozoa (r = 0.34). However, on the basis of evaluation of a single ejaculate for each stallion, the variation in these characteristics only accounted for approximately 20% of the observed variation in fertility rate.

Animals↗
PubMed1992

Increasing carbon dioxide from five percent to ten percent improves rabbit blastocyst development from cultured zygotes.

One-cell rabbit zygotes were cultured at 39 degrees C in basal synthetic medium II (BSM-II) with 5%, 10%, or 15% CO2 and humidified air to determine the effect of CO2 concentration on development in vitro. After 4 days in culture, 37% of the embryos grown in 10% or 15% CO2 had reached the hatching blastocyst stage, but only 10% of the embryos were hatching when cultured under 5% CO2 (P = 0.01). Over all blastocysts, cell numbers were 207, 246, and 205 for the 5%, 10%, and 15% CO2 treatments, respectively. In a second experiment to determine if there was a beneficial effect, particularly at the blastocyst stage, of a higher concentration of CO2, embryos were cultured 4 days in either 5% or 10% CO2 or for 2 days in 5% CO2 followed by 2 days in 10% CO2. The numbers of blastomeres per embryo and embryo diameter were greater (P < 0.05) in embryos cultured continuously in 10% CO2 or in 10% CO2 only during days 3 and 4 of culture than in embryos cultured continuously in 5% CO2. In a third experiment, one-cell rabbit zygotes were cultured with 5% or 10% CO2 in a defined, protein-free medium consisting of 1:1 RPMI 1640 and Dulbecco's modified Eagle's medium. The proportion of embryos hatching and cell counts were significantly greater (P < 0.01) when cultured in the presence of 10% CO2. These data indicate that a 10% CO2 atmosphere exerts a beneficial effect on the development of zygotes into expanding and hatching rabbit blastocysts in vitro.

Animals↗
PubMed1992

A comparison between in vitro fertilization and microinjection of immobilized spermatozoa from bulls producing spermatozoa with defects.

The objectives of this study were to compare the fertilization rate of bovine in vitro matured oocytes by in vitro fertilization (IVF) and by microinjection of a single spermatozoon (MI) and to relate these rates with fertility reported for these bulls in artificial breeding. Bull A (Holstein) had a nonreturn rate of 75%. Semen from this bull is routinely used in our standard IVF procedure. Bull B (Ayrshire), used regularly in artificial breeding and related to bull D, had a nonreturn rate of 69.2%. Bull C (Brown Swiss), with a chromosomal translocation and trisomy, achieved a nonreturn rate of 42%. Bull D (Ayrshire) produced nonmotile spermatozoa (SPZ) and had an abnormality described as "tail stump defect." No pregnancies sired by bull D have been reported. Oocytes were either fertilized in vitro by capacitated SPZ or by microinjection of a single immobilized SPZ into the ooplasm. SPZ were treated with 0.1 microM A23187 and used for IVF. For microinjection SPZ were cocultured for 5 h with bovine oviduct epithelial cells (BOEC) and then immobilized by freezing and thawing twice without cryoprotectant. A single batch of killed SPZ (stored at -25 degrees C) was used for all microinjections. All oocytes were cultured in Medium 199 for 22 h at 39 degrees C and subsequently fixed, stained, and examined for evidence of fertilization (i.e., female and male pronucleus formation, SPZ decondensation). Fertilization rates following IVF with semen from bulls A, B, C, and D were 80%, 54%, 1%, and 2%, and following microinjection were 39%, 22%, 21%, and 34%, respectively.

Animals↗
PubMed1992

Biochemical changes in bull spermatozoa during capacitation in vitro.

To evaluate the metabolic changes of bull spermatozoa (SPZ) during capacitation in vitro, SPZ were incubated for 0, 5 or 10 hours in the presence (co-culture) and absence (control) of monolayers of bovine oviduct epithelial cells, which promote capacitation-like changes in vitro. There was little change in the oxygen uptake of the SPZ after 5 hours, but after 10 hours there was a decrease, particularly in the co-cultured sample. After 5 hours there was little change in the cyclic adenosine monophosphate (cAMP) concentration of the co-culture or control SPZ, but by 10 hours the levels of cAMP decreased in both the co-cultured and control SPZ (P=0.06). The concentration of adenosine triphosphate (ATP) was somewhat decreased after 5 hours in both the co-cultured and control SPZ and the percentage of decline was much higher after 10 hours. Overall, there was no significant change in oxygen uptake or cAMP and ATP levels specifically associated with capacitation of bull SPZ.

Journal Article↗
PubMed1992

Fertilization of bovine oocytes after microsurgical injection of spermatozoa.

The objective of this study was to compare the fertilization rate of bovine oocytes matured in vitro (22, 25 or 28 hours) and in vivo (30 to 35 hours after standing estrus) following the microinjection of a single spermatozoon. A single motile spermatozoon was injected into the perivitelline space (Experiments 1 to 9), and a single immotile spermatozoon was injected into the ooplasm (Experiments 10 to 15). A single ejaculate of frozen-thawed semen was used throughout. The spermatozoa were injected either without treatment or after treatment with heparin (100 microg/ml), or Ca ionophore A23187 (0.1 microM), or co-cultured for 5 hours with bovine oviduct epithelial cells (BOEC), or they were co-cultured for 5 hours with BOEC and immobilized by freezing and thawing twice without cryoprotectant, or they remained untreated. Oocytes were placed in a droplet of hyperosmotic solution of 0.1 M sucrose in PBS to enlarge the perivitelline space (Experiments 1 to 9) or in PBS (Experiments 10 to 15). Small amounts of polyvinyl pyrrolidone (PVP) without spermatozoa were injected as a control for parthenogenetic activation. After injection, oocytes were incubated in Medium 199 for 22 hours at 39 degrees C, and they were stained with 1% aceto-orcein and examined for evidence of fertilization or parthenogenetic activation. Low rates (9 to 11%) of fertilization resulted from injection into the perivitelline space of oocytes matured for 22 hours in vitro irrespective of spermatozoa treatment. Fertilization rates were higher in oocytes matured in vivo after injection into either perivitelline space (66%) or ooplasm (74%) than in oocytes matured in vitro (9 to 44% fertilization). Surprisingly, in oocytes matured in vivo, there was no difference in the proportions fertilized by spermatozoa injection into ooplasm and parthenogenetically activated by injection of medium alone (74 and 66%, respectively).

Journal Article↗
PubMed1992

Nuclear totipotency of cultured rabbit morulae to support full-term development following nuclear transfer.

The rabbit was used as a model for nuclear transfer. A critical step in nuclear transfer is oocyte activation, which was evaluated in this research. Optimal field strength of an electric stimulus for activation was examined. A significantly higher activation rate in all criteria tested was achieved when oocytes were activated electrically with a field strength of 2.4 kV/cm versus 1.2 or 1.8 kV/cm. Also, electrical stimulation with combined alternating current (AC) and direct current (DC) was superior to DC stimulation alone for activation. In another study involving 586 oocytes, exposure of oocytes to cytochalasin B for 1 h followed by activation with electrical stimulation significantly improved development of the oocytes to blastocyst stage compared to oocytes without cytochalasin B pre-exposure (38% vs 26%, p less than 0.05). Cytochalasin B exposure alone (control), however, had no effect on activation. Exposing oocytes to activation medium without electrical stimulation also activated some oocytes. In the nuclear transfer experiment, blastomeres from 8-cell embryos cultured for 20-24 h to the 32-64-cell stage were used as nuclear donor cells. Of 491 oocytes used, 459 (93%) survived the enucleation and fusion procedure, 370 (81%) fused, and 284 (77%) developed into 2-4-cell embryos. A total of 243 of these 2-4-cell embryos were transferred to 15 pseudopregnant recipients and produced 8 young (3%). Although the efficiency is low, this study demonstrated that rabbit morulae cultured for 20-24 h to the 32-64-cell stage as nuclear donors for transfer remain totipotent.

Animals↗
PubMed1992

Synaptonemal complex analysis of a three-breakpoint translocation in a subfertile bull.

Somatic chromosome analysis of a subfertile Brown Swiss bull demonstrated a three-breakpoint translocation involving chromosomes 1, 8, and 9 in G- and R-banded karyotypes. Based on standard bovine chromosome nomenclature, the translocation was defined as t(1;8;9)(q43;q13;q26). Synaptonemal complex analysis of the chromosome aberration by electron microscopy revealed a hexavalent configuration in 52 of 53 pachytene cells. Twenty-seven cells (51%) had a completely paired hexavalent configuration showing distinctly nonhomologous pairings between normal and/or translocated chromosomes involved in the exchanges. Thirteen cells showed a hexavalent configuration with centrally unpaired chromosome segments but with completely paired terminal arms. In 13 cells (including one at zygotene) the translocation chromosomes formed an open hexavalent, and in one cell there were two completely paired trivalents. Thirty-two cells at diakinesis-MI demonstrated 28 configurations, including one large hexavalent. Testicular histology, testis size, and seminal characteristics were normal.

Animals↗
PubMed1992

Viability and acrosome staining of bull, boar and rabbit spermatozoa.

A practical and reliable staining procedure was developed to distinguish the viability and acrosomal status of bull, boar and rabbit spermatozoa. The first stain with trypan blue or Congo red is rapid and avoids artifacts. This stain is precipitated by neutral red during the 2 min required for fixation. The precipitate gives a high contrast black color, resistant to the subsequent rinsing and persists during the time required for staining the acrosome with Giemsa. Ten classes of spermatozoa are distinguished (live or dead with intact acrosomes, loose acrosomes, damaged acrosomes, no acrosome, or with no acrosome and no postacrosomal ring). The intact acrosomes are purple, the loose acrosomes are dark lavender and the damaged acrosomes are pale lavender. The anterior part of the head of live spermatozoa with no acrosome is white or light pink and the same area of dead spermatozoa is white or pale gray. The postacrosomal ring is red. The postacrosomal area of the head of live spermatozoa is white or light pink and the same part of dead spermatozoa is black, dark violet or gray. The procedure did not give satisfactory results for stallion spermatozoa.

Acrosome↗
PubMed1991

The repeatability and effect of season on seminal characteristics and computer-aided sperm analysis in the stallion.

The within-stallion repeatability and effect of season on sperm movement characteristics, determined by computer-aided sperm analysis (CASA), were compared with those of other seminal characteristics. The computer-aided determinations of sperm movement were more repeatable than the seminal characteristics of gel-free volume and sperm cell concentration based on coefficients of variation obtained from the analysis of multiple ejaculates from the same stallions. A significant (P<0.05) seasonal effect on the computer-aided movement characteristic of mean sperm linearity was observed, with a reduction in sperm linearity in the winter months. The percentage of motile and progressively motile cells and mean sperm velocity determined by CASA also tended to be lower in the winter months. These changes in sperm movement characteristics paralleled changes in other seminal characteristics. The use of CASA may have value in the potential fertility evaluation of a stallion in that it can provide relatively precise quantification of sperm movement characteristics from the evaluation of only a few ejaculates. Some CASA derived sperm movement characteristics may be lower during the physiological nonbreeding season than during the breeding season.

Journal Article↗
PubMed1991

Behavior of bull spermatozoa in bovine uterine tube epithelial cell co-culture: an in vitro model for studying the cell interactions of reproduction.

Freshly ejaculated bull semen was centrifuged and spermatozoa were resuspended in modified sperm TALP. Bovine uterine tube epithelial cell monolayers (BUTC) were obtained from cows in the periovulatory phase of estrus. In Experiment 1, sperm aliquots were assigned to culture wells containing either BUTC, BUTC-conditioned TALP, or control TALP. Sperm heads attached to the monolayers within 1 h of co-culture. Attached spermatozoa showed vigorous tail motion. At 5, 8 and 11 h of incubation at 39 degrees C, the percentage of unattached sperm cells with intact acrosome membranes and percentage of motility of these cells was measured. Sperm-BUTC co-cultures were also fixed in situ for electron microscopy. Unattached spermatozoa in co-culture had more (P<0.05) acrosomal membrane loss, showed hyperactive motion and had an overall decrease in motility as compared to sperm cells in control or conditioned medium. Evaluation by electron microscopy showed BUTC attached spermatozoa to behave in the co-culture system similar to reports for spermatozoa found in uterine tubes in vivo. Microvilli of the BUTC appeared to actively entrap the spermatozoa. Mucus-type granules could be seen on acrosomal regions and vesiculation of acrosomal membranes was seen in some cells. In Experiment 2, 43% of the 12 x 10(6) sperm cells added to 2-cm(2) BUTC bound within 4 h of co-culture. By 7 h of co-culture 19% of the previously bound sperm cells had been released from the BUTC. Released cells had limited motility and were mostly dead (73%). Sperm cells remaining on the monolayer at 7 h showed vigorous tail motion and were gradually released from the BUTC over 48 h. Spermatozoa in co-culture interacted with the BUTC in a manner much like that seen in vivo, and sperm capacitation changes were stimulated by this interaction.

Journal Article↗
PubMed1991

Factors affecting response of groups of dairy cows managed for different calving-conception intervals.

Factors which positively or adversely affect the response of cow groups to different planned conception periods were analyzed and quantified. Cows were randomly assigned to groups for breeding at postpartum intervals of 35 to 59, 60 to 90 and 120 to 150 days in a research herd, or 60 to 90 and 120 to 150 days in a cooperating commercial herd, with a planned preceding dry period of 60 days in both herds. Breeding at 35 to 59 and 60 to 90 days post partum resulted in a lower conception rate among multiparous cows but had no effect on primiparous cows. Genital disorders such as metritis, retained placenta, vulva inflammation and vaginitis, did not affect the start of luteal activity or length of the subsequent cycles, but caused a significant delay of the first behavioral estrus and, consequently, a delay of the first insemination in the earlier breeding groups. About 30% of the cows in the different categories were inseminated three or more times and were considered to be repeat breeders. Response rates (number of cows conceiving on time/number assigned) in the later breeding group improved due to higher submission rates (number of cows submitted on time/number assigned), and the conception rates were higher (number of cows conceiving on time/number submitted). The rates in all groups were adversely affected by repeat breeding and genital disorders. The latter delayed the first insemination and the conception rate.

Journal Article↗