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Biomedical subjects

R H Foote

Publications and source records attributed to R H Foote.

At least 19 recordsLinked to original sources

Within-herd use of boar semen at 5 degrees C, with a note on electronic monitoring of oestrus.

A system was designed to allow a small swine farm in a northern latitude to use its own boars for artificial insemination (AI) conveniently. Semen was collected twice weekly for 3 day use (days 0, 1 and 2), extended in an egg yolk extender and stored at 5 degrees C. Farm personnel were trained to manage the entire AI programme. For simplicity all semen collected was used for insemination. In the first test 47 gilts and 15 sows were inseminated with semen from four boars. One boar was subfertile with a farrowing rate of 36%. The averages for the other boars ranged from 71 to 100%. Then semen was collected from seven boars and all was used to inseminate 70 gilts and 55 sows with 3 x 10(9) or more sperm. Overall 63% farrowed an average of 10.1 piglets per litter. Litter size for sows was 1.5 piglets larger than for gilts. There was no difference in farrowing rate when more than 3 x 10(9) sperm were inseminated. The feasibility of initiating a complete AI programme within a small herd using herd boars was established. However, selection of the boars, use of only high quality semen, and experience with detecting oestrus was required to increase the farrowing rate. The use of various agents to protect sperm against cold shock below 15 degrees C is worthy of further investigation. A new type of electronic probe, which measures the conductivity of cervical mucus, could be helpful if a boar is not available for conventional detection of oestrus.

Animals↗

Large batch freezing of bull semen: effect of time of freezing and fructose on fertility.

Large-scale batch freezing of bull semen should be done in a processing schedule that yields the highest fertility and when it can be fitted efficiently into the work schedule. Conflicting reports have appeared on survival and fertility of bull sperm frozen within a few hours of semen collection or on the next day. To study this problem, a factorially arranged experiment with semen from 10 bulls was conducted, comparing whole milk-glycerol semen extender with and without fructose, and semen frozen in 0.5-ml straws after 4 versus 18 h of equilibration at 5 degrees C. Both fructose and 18 h of equilibration resulted in a small but significant improvement in freeze-thaw survival of sperm. A field trial followed with replicated semen collections from nine bulls processed in a whole milk-glycerol control extender frozen after 4 h of equilibration versus the addition of 1.25% (wt/vol) fructose to whole milk glycerol divided to freeze sperm after 4 and 28 h of equilibration. Semen from these bulls was used to inseminate 14,775 first-service cows. The 56-d nonreturn rates obtained for these three treatments were 74.7, 74.3, and 73.9%, respectively. As there was no difference in fertility, it would appear that programs to freeze sperm in whole milk extenders the same day of collection or the day after semen collection should yield equivalent results.

Animals↗

Sperm output and hormone concentrations in Finn and Dorset rams exposed to long- and short-day lighting.

Seasonal changes in photoperiod have a substantial effect on sexual behavior and reproduction in rams. Little information is available on sperm output from high libido versus average libido rams subjected to intensive semen collection while being exposed to controlled short versus long photoperiods. Six Finn and six Dorset rams were compared in a reversal design, which allowed rams of both breeds to be exposed to 8 h versus 16 h of light. During each of two 84-d periods rams were subjected twice to an initial depletion of epididymal sperm reserves by collecting up to 26 ejaculates of semen in 3 d, followed by up to 10 ejaculates per day, 1, 3, 5, and 7 d after the initial depletion. A total of 2673 semen samples were collected. Nearly twice as many ejaculates (63.6% of the total) were obtained from Finn rams as from Dorset rams during both the initial and subsequent 3-d sperm depletion periods. This difference in libido was associated with obtaining 33.6 +/- 3.1 x 10(9) sperm from Finn rams versus 10.0 +/- 2.2 x 10(9) sperm from Dorset rams during the initial depletion period (P<0.05). Changes in photoperiod did not affect sperm output (P>0.05) in Finn rams, but may have affected Dorset rams. With 16 h of light, prolactin was significantly (P<0.05) increased in both breeds, particularly in Finn rams. Testosterone in both breeds followed an endogenous rhythm, not affected by the change in controlled photoperiods.

Animals↗

Freezability of spermatozoa from Finn and Dorset rams in multiple semen extenders.

Ten semen extenders were tested in two experiments for cryopreservation of semen collected from four Finn and four Dorset rams. Two ejaculates of semen were combined from each ram for testing each extender treatment. The extenders consisted of a series of commonly used egg yolk-TRIS media with and without sodium and triethanolamine lauryl sulfate (STLS), a similar extender with 3-N-morpholino propane sulfonic acid (MOPS), and milk and whey extenders. In Experiment 1, extender treatments were replicated with three sets of collections from the eight rams, and in Experiment 2 with two sets. The egg yolk-TRIS-glycerol-STLS (EY(1)TSTLS) extender was significantly superior to other extenders except whole milk in protecting the sperm during freezing and thawing. In Experiment 1, a 20% egg yolk-TRIS-glycerol-STLS extender preserved 71% of the progressively motile Finn sperm (post-thaw divided by pre-freeze percentage of motile sperm), and 76% of the Dorset sperm. In Experiment 2, the corresponding values for the same EY(1)TSTLS extender used with Finn and Dorset sperm were 86 and 64%, respectively. Without STLS the egg yolk extenders were significantly less effective in protecting cryopreserved ram sperm. This egg yolk-TRIS extender, containing STLS and glycerol, may hold promise for freezing ram sperm that could be used successfully for intracervical insemination.

Animals↗

Blood catalase and haematocrit values in a breeding colony of Dutch-belted rabbits.

Rabbit seminal plasma catalase is much higher than in the semen of other mammals, and differences appear to be inherited. Because of the scarcity of information on rabbit blood catalase and haematocrit in Dutch-belted rabbits, an investigation of possible effects of gender, age and genetics on these variables was undertaken. There were 191 rabbits sampled at 2-3 months, 130 at 12 months and 61 at 18-24 months of age. There was no age effect on the haematocrit values and on blood catalase activity. At 12 months of age males had an average haematocrit value of 44% compared with 40% for females (P < 0.05). Corresponding average catalase values were 431 and 356 units/ml of blood (P < 0.05). Also catalase was measured in the semen and blood of 34 males, and males differed in both their blood and semen catalase activity (P < 0.05). The correlation between the two traits was r = 0.44. Heritability (h2) estimates, based on 231 rabbits were 0.40 for blood catalase activity, and 0.26 for haematocrit. The genetic correlation between the two variables was 0.83 (P < 0.05). These studies are consistent with the literature in that female rabbits have a slightly lower haematocrit value than males, and this is associated with a lower catalase activity. This appears to be the first report of a study that compares rabbit blood catalase in males and females of different ages. Preliminary evidence that differences may have a heritable basis is consistent with previous studies on rabbit semen catalase.

Age Factors↗

Inhibition of sperm motility does not affect live-dead separation of bull sperm by glass beads.

AIM: This study was designed to explore factors which influence binding of dead versus live sperm to glass filters. METHODS: Multiple semen collections from bulls were used to explore selective filtration of bull sperm as influenced by nonlethal inhibition of sperm motility with fluoride, killing of sperm by quick-freezing, alteration of the glass surface with silicone, and different intervals of sexual rest between semen collections. RESULTS: A comparison of glass spheres 100, 200 and 390 microm in diameter indicated that 200 microm spheres were optimal for selective filtration. Quantitative separation of live from dead sperm was demonstrated with a correlation between the percentage of motile sperm and retention of sperm by the filter of r = -0.87 (P < 0.05). Up to 0.02 mol/L NaFl did not alter the proportion of sperm retained by the filter despite inhibiting sperm motility during filtration, an inhibition which was reversible. Proportions of live-dead sperm, based upon eosin staining, were unaffected by fluoride. Coating the glass spheres with silicone greatly reduced selective filtration. Dead sperm adherence to glass was reduced and resistance to NaFl inhibition was increased by daily ejaculation versus one-week intervals of sexual rest. CONCLUSION: These studies indicate that the adherence of sperm to glass is primarily due to some form of physico-chemical change accompanying death of the sperm cell independent of active sperm motility. This attraction between the sperm plasma membrane and glass is modified by the age of the ejaculated sperm. This information is useful in evaluating different clinical procedures used for sperm separation.

Animals↗

Importance of inseminating only cows in estrus.

Reproductive efficiency is a key component in successful dairy farm management. A study was initiated to evaluate the incidence and consequences of inseminating dairy cows in the middle of the estrous cycle or while pregnant. In a research herd of 242 Holsteins, managed for reproduction under typical farm conditions, milk progesterone (P4) was assayed 3 times per wk for at least 120 d postpartum. The P4 cycle was compared with the estrus detection and breeding records and pregnancy diagnosis 6 wk after insemination. About 19% of the inseminations were performed when P4 was high in the estrous cycle and in pregnant cows. Insemination of pregnant cows led to an estimated 17% induced embryonic death or abortion. In Israel, inseminators are extensively trained to detect cows not in estrus. They reject about 16% of the cows submitted for reinsemination, with a 95% accuracy of rejection of 44% of the cows that were pregnant. The pattern of submission of cows for reinsemination in areas around New York State was similar to Israel and to the experimental herd. These results indicate that more careful submission and rejection can reduce the unnecessary use of semen, reduce abortions and minimize long calving intervals, all contributing to the success of a dairy herd operation.

Abortion, Veterinary↗

Allocation of inner cell mass and trophectoderm cells to the preimplantation blastocyst of the domestic ferret, Mustela putorius furo.

The growth of ferret preimplantation blastocysts in vivo, collected between 156 and 240 hr post coitum, was investigated. A technique, combining immunosurgery and differential fluorochrome staining, was used to discriminate between inner cell mass (ICM) and trophectoderm (TE) cells. Using the stains propidium iodide and bisbenzimide (Hoechst 33342), the ICM was stained blue and the TE was stained pink. The ICM and TE counts for 90 blastocysts, respectively, averaged 25 and 63 at 156 hr and increased exponentially to 2077 and 4137 at 240 hr. The Box-Cox procedure was used for choosing a transformation that minimized the error sum of squares for a linear regression of Y (cell count) on X (time in hr). Logarithmic transformations of the ICM, TE and total cell count gave a good fit, but the following equations obtained by the Box-Cox procedure provided the best fit, where Y is cell count and X is time in hours. For inner cell mass: Y = [(176.06 + 2.45X)/-899.44 + 1]-3.33; trophectoderm: Y = [(301.38 + 14.48X)/-6863.42 + 1]-10; and total: Y = [(2266.97 + 17.0X)/-7837.21 + 1]-5. The R2 values were 0.73, 0.84, and 0.84, respectively. The exponential growth of the ferret embryo during the time interval that measurements were made fits the general pattern described for other mammalian embryos. This report is the first to characterize the pattern of cell allocation and growth in preimplantation blastocysts of the ferret, and the first such report for a carnivore. The pattern of in vivo development provides a standard for judging the quality of in vitro produced and matured ferret embryos and, concomitantly, a means to evaluate culture systems.

Animals↗

Bull sperm surface "craters" and other aspects of semen quality.

Semen from 200 Holstein bulls in an artificial insemination center was examined for the frequency of craters on the surface of sperm heads, as visualized with the aid of differential interference contrast microscopy. Semen from 100 of these bulls was examined in more detail in 2 experiments by staining with eosin-aniline blue to determine the relationship of unstained spermatozoa, and spermatozoa with normal acrosomes with apical ridges to the incidence of craters and fertility. Only 3 of 100 bulls had a substantial incidence of craters (15 to 23%), whereas the average of the other 97 bulls in 2 experiments was 1 to 3%. The percentage of sperm cells with craters was correlated (P < 0.05) with the percentage of unstained spermatozoa (r = -0.29 and sperm cells with normal acrosomes (r = -0.52) but was not significantly correlated (r = -0.24) with the nonreturn rate. One bull with many sperm cells with craters was slaughtered, and the epididymal spermatozoa were examined. The high incidence of sperm cells with craters was limited to one side, with the testis on that side having 2 Sertoli cell tumors. The remaining 2 bulls as well as one other that produced 16% of sperm cells with craters did so only temporarily. Within a few months crater sperm production had decreased and semen quality increased. The condition usually appears to be transitory, presumably due to temporary stress.

Animals↗

Evaluation of sperm tail membrane integrity by light microscopy.

During routine evaluation of trypan blue-Giemsa stained semen smears, sperm cells can be found with unstained heads and with stained tails. It was hypothesized that these cells were immotile and should not be considered as live. Sperm motility was determined in isoosmotic, and presumably isotonic trypan blue-stained wet preparations. Bull, ram and boar semen smears were stained with hypoosmotic trypan blue-Giemsa to compare the relationship between the percentage of stained sperm tails and the percentage of sperm tails remaining straight under hypoosmotic conditions. Actively moving spermatozoa with unstained heads, but with stained tails were never observed in wet preparations. The correlation coefficient found between the percentage of sperm with stained tails and the percentage with straight tails was 0.81, 0.94 and 0.85 for bull, ram and boar spermatozoa, respectively. Results of this study show that sperm cells with an intact head membrane, but a stained and presumably membrane-damaged tail are not motile. Therefore these cells should be included in the dead category rather than alive in the usual live-dead studies with vital stains.

Animals↗

Nonsurgical collection and nonsurgical transfer of preimplantation embryos in the domestic rabbit (Oryctolagus cuniculus) and domestic ferret (Mustela putorius furo).

The objective of this study was to develop nonsurgical methods of embryo collection and transfer in domestic rabbits (Oryctolagus cuniculus) and domestic ferrets (Mustela putorius furo) to serve as models for use in mammals in which surgical procedures are the usual means for applying embryo transfer technology. Specially designed transcervical catheters were used together with a fibre optic endoscope to visualize and then catheterize the rabbit and ferret cervices. Five consecutive transcervical uterine flushes in each of eight superovulated female rabbits 78-89 h after an ovulatory injection of LH resulted in the retrieval of 187 embryos, for an average of 23 embryos per rabbit. A total of 116 embryos were nonsurgically transferred to the uteri of ten recipients, and resulted in 23 young (20%). Eight rabbits (80%) produced young with an average litter size of 2.88 (range 1-7). Ten consecutive transcervical uterine flushes in each of 37 female ferrets 145-178 h after an ovulatory injection of hCG resulted in the retrieval of 324 embryos, an average of 8.76 embryos per ferret. A total of 251 embryos from 27 donors were nonsurgically transferred to the uteri of 31 recipients, and resulted in 65 young (26%). Twenty-eight of the recipients (90%) were initially pregnant, as indicated by postpartum necropsies, and twenty-two ferrets (71%) produced young. The average litter size was 2.95 (range 1-7). This is the first report of live births resulting from the nonsurgical collection of embryos from a donor followed by nonsurgical transfer of those same embryos to a synchronous recipient. The methods reported here can serve as models for use in other mammals in which direct visualization and manipulation of the cervix are not possible, and will be particularly useful in endangered species.

Animals↗

Gonadotropin-releasing hormone improves reproductive performance of dairy cows with slow involution of the reproductive tract.

Eighty multiparous Holstein cows were assigned randomly at calving to receive either 100 microg of GnRH or saline 13 or 14 d postpartum (PP). From 4 to 28 d PP the cows' reproductive organs were palpated weekly per rectum, and cows were subclassified within each group as undergoing slow (delayed) cervical and uterine involution (abnormal) or as normal cows. Last milk obtained after removing the milking machine was assayed for progesterone 3 times a week for 120 d PP. Fourteen of the 80 cows were removed from the experiment because of culling or various veterinary treatments of pathologic conditions that could confound analysis of the GnRH treatment effects. As expected, the treatment of normal cows with GnRH had no significant effects on the first estrus or the first estrous cycle PP, on services per conception, days open, or any other reproductive trait measured. However, in the abnormal group of cows receiving saline, first rebreeding after calving was delayed (81 vs. 67 d), fewer were pregnant by 105 d PP (23 vs. 64%), and number of days open was greater (121 vs. 87 d) compared with those receiving GnRH; all were significant (P<.05). Treated abnormal cows were equivalent to the control normal cows. Thus, GnRH given 13 to 14 d PP to cows characterized as undergoing slow involution of the reproductive system, but with no other clinical problems, seems to assist in promoting rapid normal reproductive function. Subsequent losses due to culling were greatly reduced.

Animals↗

Fertility of bull sperm frozen and stored in clarified egg yolk-Tris-glycerol extender.

Egg yolk-Tris-glycerol extender, which is widely used in commercial artificial insemination (AI), was modified by replacing the 20% egg yolk (vol/vol) with a supernatant from egg yolk centrifuged at 50,000 x g for 2 h and then used to cryopreserve bull sperm. Preliminary studies showed that 20% egg yolk interfered with biochemical assays, which could be overcome by centrifugation. In Experiment 1, semen from 4 Holstein bulls was frozen in the experimental Tris extender and compared with the whole milk-glycerol control. A total of 2256 first services resulted in 72.6% 60- to 90-d nonreturns for the control and 71.0% for the Tris extender. In Experiment 2, semen from 10 Holstein bulls was frozen in the experimental Tris extender. Half of the semen was used immediately and half was stored in liquid nitrogen for 1 yr before distribution. The nonreturn rates based on 8878 first services for 7 bulls that completed both parts of the trial were 70.9% initially and 71.6% 1 yr later. This time trend difference of 0.7% was comparable to 0.6% for the AI mean for Holstein sires used at the same time. These fertility results and previous laboratory studies indicate that the conventional egg yolk-Tris-glycerol might be simplified for cryopreserving bull sperm. The experimental Tris extender also was suitable for making biochemical measurements.

Animals↗

Short communication: an electronic probe versus milk progesterone as aids for reproductive management of small dairy herds.

A simple probe especially designed to take electrical resistance measurements at different positions in the anterior vagina of a cow was compared with milk progesterone determinations on 108 cows. Milk samples were taken 3 x weekly, 21 to 60 d postpartum, at the time of insemination, and 21 to 23 d later. Electrical resistance measurements were made on a similar schedule. In 10 other herds, 187 cows had only milk samples taken. No cows with high milk progesterone values became pregnant when inseminated, but the electrical resistance values were less accurate in designating which cows were suitable or unsuitable for insemination. Both low milk progesterone and low electrical resistance values 21 to 23 d after insemination provided an early and accurate indication of a need for reinsemination. These indicators were consistent with 94 to 100% of these cows being diagnosed as not pregnant 6 to 8 wk later. Daily probing, starting about 19 d after a previous insemination, could serve as an early check of pregnancy and assist in identifying cows for immediate reinsemination.

Animals↗

Resazurin reduction and other tests of semen quality and fertility of bulls.

AIM: This study was undertaken to compare the reduction in color of two dyes methylene blue (MBRT) and resazurin dye (RRT) with other tests of bull semen quality and to examine their relationship to fertility. METHODS: One hundred sixty-four ejaculates from 59 bulls were examined, processed, and used for 30,016 inseminations. RESULTS: Bulls used in artificial insemination have been selected for high semen quality and fertility, and semen from these bulls averaged 80.6% unstained sperm, only 11% had abnormalities, and fertility ranged from 64% to 76%. The MBRT and RRT were run with standardized sperm numbers to prevent sperm concentration from influencing the dye reduction time. A short RRT was correlated with higher fertility (r = -0.26) but MBRT was not correlated (r = -0.05, P > 0.05). The correlations were low, but are typical and reflect the statistical effect of the large binomial variance associated with pregnancy or nonpregnancy being coded as 1 or 0. CONCLUSION: The fact that the RRT was significantly correlated with fertility when sperm numbers were standardized, and published reports that resazurin is useful for monitoring other semen characteristics, indicate that this dye may provide valuable information not given by other simple laboratory tests.

Animals↗

Osmotic effects on volume and motility of bull sperm exposed to membrane permeable and nonpermeable agents.

Factorially arranged experiments were designed to study prefreeze packed cell volume (PCV) changes and associated percentages of motile and unstained bull sperm in simple macromolecule-free Tyrode's solution and egg yolk-Tris (EYT), varying in osmolarity, and with addition of rapidly permeating cryoprotectants, glycerol and 1,2-propanediol, and nonpermeating substances, sucrose and NaCl. The percentage of motile and unstained sperm was assessed after resuspending sperm in 300 mOsm/L Tyrode's solution. At 25 degreesC PCV increased in Tyrode's solution as osmolarity was decreased from 250 to 150 mOsm/L and decreased as Tyrode's solution was increased to 400 mosmol/L. The relationship of PCV to the reciprocal of the osmolarity was essentially linear over the range of 150 to 400 mOsm/L, but PCV did not decrease further in solutions ranging from 500 to 1000 mOsm/L. The percentage of motile sperm declined to zero in Tyrode's solution at 700 mOsm/L, but 40% of the sperm were still unstained in 1000 mOsm/L solutions. The addition of glycerol or 1,2-propanediol had little effect on PCV. With glycerol or 1,2-propanediol added to 308 mOsm/L Tyrode's solution to give a total of 1267 mOsm/L, there were 49 and 56% motile sperm, respectively, compared to 1% with NaCl added to give 787 mOsm/L. The PCV and percentage of motile sperm suspended in EYT responded to osmotic changes similar to those reported for Tyrode's solution at both 25 and 5 degreesC. Some sperm remained motile after initial exposure to 800 mOsm/L solutions. These findings may have application in improving bull sperm cryopreservation.

Animals↗

Survival of bull sperm frozen at different rates in media varying in osmolarity.

The effects of freezing procedures, osmolarity, trehalose, and sucrose on survival of bull sperm in whole milk (WM) and egg yolk-Tris (EYT), semen extenders used worldwide, were studied. Sperm were added to extenders at 25 degreesC, cooled slowly to 5 degreesC, glycerolated, packaged in 0.5-ml straws, and frozen. Different freezing rates were accomplished in two steps. Straws were transferred from +5 degreesC to nitrogen vapor at temperatures ranging from -10 to -100 degreesC in the first step and to liquid nitrogen in the second step. Straws were thawed in water at 35 degreesC. A substantial decrease in sperm motility occurred between -10 and -20 degreesC, as abrupt nucleation occurred following supercooling to -13 degreesC. To study the interactions between osmolarity x cooling rate, WM and EYT extenders were prepared to yield media measuring 220 to 420 mOsm/L. The optimal first-step range of cooling in the two-step procedure was -30 to -70 degreesC, and the highest proportions of motile sperm after freezing and thawing were 61 to 62 in 260 to 300 mOsm/L WM and 63 to 64% in 300 to 340 mOsm/L EYT, equivalent to the results with the control procedure used commercially. As the cooling rate increased (first step to -100 degreesC) sperm motility was much higher in hypertonic than in hypotonic extenders (P < 0.05), indicating the importance of partial dehydration before rapid cooling. Replacing part of EYT and WM with equivalent solutions (same mOsm/L) of sucrose or trehalose had no appreciable effect. These results provide a basis for further investigating simple freezing systems that might be more effective in preserving bull sperm than those currently available.

Animals↗