Showing papers on "Semen published in 1983"

Unique seminal quality in the South African cheetah and a comparative evaluation in the domestic cat.

Abstract: Analysis of 40 semen samples collected by electroejaculation from 18 cheetahs revealed no major differences in seminal traits among Transvaal, South West (Namibia) or hybrid (Transvaal X South West) males. However, mean spermatozoal concentration (14.5 X 10(6) spermatozoa/ml of ejaculate) and percent motility (54.0%) were less in cheetahs than in domestic cats (147.0 X 10(6) spermatozoa/ml of ejaculate, 77.0% motility) subjected to the same electroejaculation regimen. On the average, cheetah ejaculates contained 71.0% morphologically abnormal spermatozoa compared to 29.1% aberrant spermatozoal forms in the domestic cat. These results indicate that seminal characteristics in the cheetah are markedly inferior compared to the domestic cat, particularly with respect to the incidence of pleiomorphic spermatozoa. Because a recent parallel study demonstrates that the cheetah lacks genetic variation, it appears likely that spermatozoal abnormalities are a genetic consequence of genomic homozygosity characteristic of this endangered species.

Effects of osmolality and potassium on motility of spermatozoa from freshwater cyprinid fishes

Abstract: Spermatozoa of freshwater Cyprinidae (goldfish, carp, crucian carp and dace) remained immotile when the semen was diluted in solutions of NaCl, KCl, mannitol or glucose iso-osmolar to the seminal plasma (300 mosmol kg-1). The spermatozoa became motile in media containing these solutes if the osmolality was lower than that of the seminal plasma, suggesting that motility is suppressed by the osmolality of the seminal plasma in the sperm duct and initiated by a decrease of osmolality upon spawning into fresh water. Potassium was a major component of seminal plasma, having a concentration 20-30 times higher than that in the blood plasma in goldfish and carp. Sodium concentration in seminal plasma was lower than that in blood plasma. Potassium increased viability and speed of sperm movement at a concentration below that in the seminal plasma, whereas sodium and the nonelectrolytes were less effective. Potassium released with spermatozoa at spawning may therefore stimulate motility which has already been initiated by the decrease of osmolality.

Quantification of the X- and Y-chromosome-bearing spermatozoa of domestic animals by flow cytometry.

Abstract: The relative content of DNA in spermatozoa presumed to be the X- and Y-chromosome-bearing gametes from bulls, boars, rams and rabbits and the amount of DNA in spermatozoa of cockerels was determined by flow cytometry. Differences in the relative content of DNA and proportions of the presumed X- and Y-sperm populations in cryopreserved semen from Holstein, Jersey, Angus, Hereford and Brahman bulls were also determined. Spermatozoa were washed by centrifugation using a series of dimethyl sulfoxide solutions made in isotonic sodium citrate, fixed in ethanol, treated with papain and dithioerythritol to loosen the chromatin structure and remove cellular organelles, and stained quantitatively for DNA with the fluorochrome 4'-6-diamidino-2-phenylindole (DAPI). Approximately 5000 stained sperm nuclei, which were nonviable due to the removal of other cellular organelles during the washing procedure, were measured for DNA in an epi-illumination flow cytometer. A single distinct peak for cockerel spermatozoa and two symmetrical, overlapping peaks for species with X- and Y-spermatozoa were seen. This and other evidence strongly supports the interpretation that the peaks represent the X- and Y-sperm populations. The content of DNA in sperm nuclei from cockerels, bulls, boars, rams and rabbits, as determined by fluorescence flow cytometry, corresponded to biochemical estimates of DNA per sperm cell. Analyses of the bimodal histograms by computer-fitting two Gaussian distributions to the data showed the means of the peaks differed by 3.9, 3.7, 4.1 and 3.9% for bulls, boars, rams and rabbits, respectively. In four replicate analyses of semen from 25 bulls representing 5 breeds, the average population of sperm nuclei in the Y-peaks ranged from 49.5 to 50.5% for all breeds. The X-Y peak differences did not vary within each breed, but were significantly different when the breeds were compared. Spermatozoa from Jersey bulls had larger X-Y peak differences (P less than 0.001) than spermatozoa from Holstein, Hereford, and Angus bulls; spermatozoa from Brahman bulls had smaller X-Y differences (P less than 0.004). It is suggested from the evidence obtained in these studies that flow cytometry can be used to assess the proportion of X- and Y-spermatozoa in semen of domestic animals and is thereby applicable to verification of the effectiveness of enrichment techniques for X- or Y-spermatozoa.

Effects of potassium and osmolality on spermatozoan motility of salmonid fishes

Abstract: In salmonid fishes, rainbow trout and masu salmon, and the plecoglossid fish, ayu, seminal plasma had an osmolality around 300 mosmol kg-1 isotonic to the blood plasma, and contained a higher concentration of potassium than the blood plasma. Spermatozoa of salmonid fishes were motile when semen was diluted 1:100 with solutions of sodium chloride or mannitol, over the osmotic range of 0–300 mosmol kg-1. They were immotile in sodium chloride solution containing several mM potassium. This indicates that osmolality is not an essential determinant of sperm motility in the Salmonidae, and that sperm motility in these species is suppressed by the seminal potassium in the sperm duct, and initiated by a decrease in potassium concentration surrounding spawned spermatozoa when they are released into fresh water.

Isolation of motile spermatozoa by density gradient centrifugation in Percoll

Abstract: A procedure using centrifugation in density gradients composed of Percoll was developed for isolation of spermatozoa from mammalian semen. To evaluate the technique, rabbit, human, or bovine semen was layered over continuous Percoll gradients ranging in density from 1.02 to 1.13 gm/ml and centrifuged at 1,500g for 45 min. After centrifugation, the seminal plasma remained above the gradient, whereas the spermatozoa and seminal particles were distributed within the gradient according to their buoyant densities. Unlike most washing techniques, no sperm pellet was formed; instead, the spermatozoa were concentrated into a compact band above the most dense layer of Percoll. The spermatozoa recovered from the gradient were easily resuspended by gentle techniques. Thus, the mechanical stress to the spermatozoa was minimized. Osmotic stress to the spermatozoa was also negligible as the Percoll gradients were isotonic throughout. Spermatozoa obtained by this technique possessed motility equivalent to that of spermatozoa in the unfractionated semen. Sperm suspensions recovered from the gradients contained less than 5% of the nonspermatozoal particles present in the original samples of unfractionated semen. Soluble seminal components were also efficiently removed from the spermatozoa. Thirty billion bovine spermatozoa could be fractionated on a single gradient without loss of effectiveness. Recovery of spermatozoa from these preparative separations averaged 80%. These results demonstrated that Percoll was a superior medium for efficient density gradient isolation of motile spermatozoa free of contamination by other seminal constituents.

Semen Analysis in Testicular Cancer and Hodgkin's Disease: Pre‐ and Post‐Treatment Findings and Implications for Cryopreservation

Abstract: Summary— Over a 7–year period, seminal analysis has been performed on 208 patients with testicular tumours, after orchiectomy, but before any other treatment. Only 22% of 54 patients with seminomas, and 29% of 154 patients with teratomas or mixed tumours, had sperm counts exceeding 10 million per ml. Very low sperm counts were observed in some patients who had previously fathered children. Post-treatment sperm counts were done in 117 patients, 80 of whom had received multiple drug chemotherapy: 42 of these men had pre- and post-treatment sperm counts. Overall, 24% of men receiving chemotherapy recovered sperm counts greater than 10 million per ml up to 3 years after therapy. Surprisingly, such recovery was seen in 35% of 23 men with initially poor sperm counts, but in only 26% of 19 with good initial counts. Only 27% of 49 patients with Hodgkin's disease had initial sperm counts of more than 10 million per ml; after chemotherapy only 1 of 29 patients recovered to this level. Only one quarter of these young men had semen which was adequate for cryopreservation. Artificial insemination with semen preserved in liquid nitrogen has been performed in 15 couples: 2 normal babies have been produced and a third pregnancy is progressing normally.

Has the fertility of Danish men declined through the years in terms of semen quality? A comparison of semen qualities between 1952 and 1972.

Abstract: The semen analysis of 1,077 men examined in 1952 was compared with the semen analysis of 1,000 men examined in 1972 in order to assess if fertility in Danish men has declined during the period. The men on both occasions were examined because of a fertility problem. In 1952 6.2% of the men had azoospermia compared with 3.9% in 1972 (P less than 0.05). Between 1952 and 1972 there was a fall in sperm count (P less than 0.01, median 73.4, and 54.5 mill/ml), a deterioration in spermatozoa motility (P less than 0.001), an increase in number of abnormal spermatozoa (P less than 0.01, median 26.0%, and 44.8%), and a deterioration in fertility class according to the Hammen system (P less than 0.001). Semen volume and number of immobile spermatozoa did not change. This study of Danish men, like studies from other industrialized countries, indicates a fall in semen quality and male fertility since the early fifties.

Rapid analysis of normal and abnormal cell types in human semen and testis biopsies by flow cytometry.

Abstract: A simple, rapid procedure is described that quantitates RNA content and DNA content/chromatin condensation for each of many possible cell types and differentiation levels of the cells present in human semen. A fresh semen sample (1-6 hr postemission) or frozen sample (allowing samples to be accumulated and sent to a laboratory) is treated with a detergent solution, stained with acridine orange (AO), and measured by flow cytometry (FCM); approximately 10 minutes are required to measure 5,000 cells per sample and analyze the data with computer assistance. The following can be learned from a single measurement: a) the percentage of each cell type in semen including, i) mature sperm, ii) immature sperm precursor cells, representing all stages of development from spermatogonia to mature sperm, iii) somatic cells, e.g., leukocytes; b) normality/abnormality of sperm nuclear chromatin condensation. These measurements can be correlated with cell types in testis biopsies identified by two-parameter FCM measurements (RNA versus DNA) using AO as the fluorescent probe.

Movement characteristics and acrosomal status of rabbit spermatozoa recovered at the site and time of fertilization.

Abstract: Rabbit spermatozoa were recovered from the oviductal ampullae 11 h postcoitus by an oil microflush technique. Their movement was evaluated in the ampullar fluid, or in ampullar fluid diluted with in vitro fertilization medium, in slide preparations which were approximately 25 micron or 100 micron deep. The movement of these sperm was compared with the movement of ejaculated sperm in diluted semen. Movement parameters measured from videotapes recorded by a high-speed camera were coded according to treatment and entered into a microcomputer for statistical analysis. A total of 157 spermatozoa were recovered from the oviducts of 16 does: 152 were motile and 126 were free-swimming. Nearly all of the free-swimming sperm swam in trajectories whose average paths were circular. The flagellar beat pattern of the circular swimmers was asymmetric and nearly planar, and the sperm did not roll. Spermatozoa observed in 25-micron slide preparations produced smaller flagellar bends than sperm swimming in 100-micron preparations and tended to swim in larger circles which were oriented in the plane of the slide. Spermatozoa observed within the cumulus matrix moved in a slow, erratic, sinuous manner, but resumed rapid circling upon leaving the matrix. It was concluded that the ampullar sperm were hyperactivated, retaining this physiological condition as they entered the cumulus. The movement qualitatively resembled that of hyperactivated guinea pig and hamster spermatozoa because these species effectively swim in circles. In contrast, 80% of the ejaculated spermatozoa swam in linear trajectories, resulting from relatively symmetrical, flagellar beat patterns. The percentage of rolling spermatozoa and the rolling frequencies were less in the 25-micron than the 100-micron slide preparations. Thus, the movement parameters of both ampullar and ejaculated spermatozoa were affected by the geometry of their observation chambers. This influence should be taken into account when observing sperm motility in vitro. It could also be important in vivo, where changes in sperm movement in response to epithelial surfaces might provide an advantage for reaching the cumulus mass. Ninety-eight percent of the motile ampullar sperm were observed to have acrosomes, including all spermatozoa found within the cumulus matrix.

Errors Inherent in the Performance of a Routine Semen Analysis

Abstract: Summary— A group of 26 medical laboratory technicians and pathologists performed a semen analysis on an aliquot of the same sample of semen The mean sperm concentration obtained was 467 ± 106/ml ± 401 (2 sd) with a range of values lying between 10 and 98 ± 106/ml and giving a coefficient of variation of 443% Similar wide ranges were seen in the visual estimation of percentage motility and abnormal morphology Results obtained from three technicians, all highly experienced in performing semen analyses, produced a mean sperm concentration of 66 times 106/ml with a coefficient of variation of 378% When the sperm concentration in semen was low, replicate analyses of four separate semen specimens produced coefficients of variation that were unacceptably high

Sperm morphology assessment as an indicator of human fertilizing capacity.

Abstract: Semen samples from 95 men were examined by routine semen analysis and specific histologic staining for sperm morphology. The men were classified into fertile and infertile groups on the basis of clinical evaluation and in vitro testing, using the zona-free hamster egg penetration assay. Thirty men were designated as fertile, as they had fathered children and their sperm showed penetration of greater than 20% of the zona-free hamster eggs with the in vitro fertilization test. Subjects classified as infertile were men from infertile couples whose wives showed no evidence of infertility and whose in vitro fertilization ability was 10% or less. The semen analysis parameters of the fertile and infertile groups were significantly different. Fertile men had mean values of 108 X 10(6) sperm/ml, 61% motile, 64% normal forms (sperm with oval morphology), and 69% penetration in vitro. The mean values for infertile men were significantly lower: 42 X 10(6) sperm/ml, 45% motile, 32% normal forms, and 3.2% penetration in vitro. The importance of the morphology parameter was revealed by comparison of the percentage of penetration with count, motility, and morphology. Penetration correlated best with morphology (r = 0.730) as compared with motility (r = 0.451) and count (r = 0.605). The distribution of abnormalities in the infertile group revealed 81.6% with abnormal morphology (less than 50%), 53.8% with abnormal motility (less than 50%), and 38.5% with abnormal count (less than 20 million/ml). As a single parameter, decreased number of normal forms appears to be a good indicator for clinical infertility if in vitro fertilization testing is not available.

Fertilizing ability of cock spermatozoa from the testis epididymis and vas deferens following intramagnal insemination.

Abstract: A reexamination of the fertilizing ability of cock spermatozoa from the testis, epididymis and vas deferens was accomplished through the use of intramagnal insemination. Intramagnal insemination of spermatozoa taken from the testes, epididymides and vas deferentia resulted in fertility levels of 85-90% during the first week and levels of 67-90% the second week after a single insemination. In contrast, vaginal insemination of testicular spermatozoa resulted in a total absence of fertile eggs. Vaginal and intrainagnal insemination of ejaculated spermatozoa resulted in fertility levels of 70 and 71%, respectively, during the first week and levels of 42 and 53%, respectively, the second week after a single insemination. There were no significant differences in hatchability of fertile eggs from hens inseminated with semen from different regions of the male tract or by different routes of insemination.

Influence of caffeine on movement characteristics, fertilizing capacity and ability to penetrate cervical mucus of human spermatozoa

Abstract: Summary. When added to frozen\p=n-\thawedhuman semen, the 3 doses of caffeine tested (2, 5 and 10 mM) induced a significant increase in the percentage of motile spermatozoa but did not influence the quality of movement. Considerable variability was noted between samples in their responsiveness to caffeine which, at the 5 and 10 mM doses, was significantly correlated with the degree of motility lost during cryostorage. Caffeine treatment of frozen\p=n-\thawedhuman spermatozoa also increased the number of spermatozoa penetrating cervical mucus in unit time, by increasing the frequency rather than the success of collisions between spermatozoa and the cervical mucus interface. When caffeine-stimulated spermatozoa were washed free of seminal plasma containing this compound they were no longer at an advantage with respect to their motility or fertilizing ability. When 2 mM-caffeine was added to washed suspensions of capacitated spermatozoa it failed to stimulate motility but did significantly enhance the fertilizing ability of the spermatozoa, indicating a possible clinical role for this compound in in-vitro fertilization therapy.

Effects of Occupational Exposure to Lead on Sperm and Semen

Abstract: Semen qualities were studied in samples from workers exposed to lead in a factory for storage batteries. The study included two groups with different degrees of exposure. The mean lead-blood concentration (Pb-B) values for the men in the two groups during the six months preceding the study were 45 µg/100 ml and 22 µg/100 ml, respectively. Each man delivered at least one semen sample during a test period and most of the men participated in both test periods, which took place within a six-month interval. The size of each of the four groups, was 16 or more men. The semen qualities assessed included sperm count, sperm motility (qualitative and quantitative), sperm morphology, sperm chromatin stability when exposed to sodium dodecyl sulphate, and release of LDH-X into the seminal plasma. The secretory function of the prostate was assessed by analyses of such markers as acid phosphatase, zinc, and magnesium. Fructose was used as the marker for the secretory function of the seminal vesicles. All semen samples had values within the expected limits for that population. A subtle but significant difference was found between the two groups for sperm chromatin stability, indicating that the exposure to lead had decreased the stability of the spermatozoa. Moreover, a decreased secretory function of the accessory genital glands was noted more frequently among the men with the higher degree of exposure than among those in the other group. For all other semen variables, there were no differences between the groups.

Fertility of Fresh and Frozen -Thawed Semen of the Angora Goat

Abstract: In six experiments, involving a total of 954 does, fresh and frozen-thawed semen processed in a Tris-based diluent was used for artificial insemination,

Fertility of unfrozen and frozen stallion spermatozoa extended in EDTA-lactose-egg yolk and packaged in straws.

Abstract: The fertility of frozen-thawed semen was compared with that obtained using fresh semen extended in skim milk. Semen for freezing was obtained in June from four stallions of unknown fertility; two ejaculates were collected 1 to 2 h apart every 3 or 4 d. The gel-free fraction of the ejaculate was mixed 1:1 with a glucose-EDTA solution (disodium ethylenediaminetetraacetic acid) and centrifuged at 650 x g for 15 min. The spermatozoa were resuspended in an EDTA-lactose-egg yolk extender containing 5% glycerol. Semen was frozen in .5-ml French straws containing 250 x 10(6) progressively motile spermatozoa before freezing. Only for 31% of the 54 ejaculates frozen was post-thaw spermatozoal motility greater than or equal to 50% of the percentage of progressively motile spermatozoa observed during evaluation of the neat semen. Spermatozoa in second ejaculates apparently were more susceptible to the adverse effects of dilution and centrifugation than spermatozoa in first ejaculates. Only samples containing greater than 30% progressively motile spermatozoa after freezing and thawing (at 38 C) were used for insemination. In June and July, 101 mares were inseminated daily with semen from one of three stallions beginning on d 2 and continuing through the end of estrus for one cycle. Mares were inseminated with semen in one straw or with 250 x 10(6) progressively motile spermatozoa extended in 10 ml of skim milk. Because of the poor survival of spermatozoa that had been frozen and thawed, mares inseminated with frozen-thawed semen received 100 to 130 x 10(6) progressively motile spermatozoa as compared with 250 x 10(6) progressively motile spermatozoa for mares inseminated with fresh semen. One cycle pregnancy rates, based on rectal palpation 50 to 60 d after ovulation, were 29% using frozen-thawed semen and 66% using fresh semen (P less than .05). Values for individual stallions were 19, 24 and 47% with frozen semen and 47, 61 and 67% with fresh semen. Routine use of frozen stallion semen is not recommended at this time.

A study of factors affecting the success of human fertilization in vitro. II. Influence of semen quality and oocyte maturity on fertilization and cleavage.

Abstract: The incidence of in vitro fertilization was analyzed with respect to the degree of cumulus dissociation (expansion) at the time of oocyte recovery and also the semen quality. Of the oocytes surrounded by perfectly ("++") or moderately ("+") dissociated cumuli, 78.6% and 30.8%, respectively (P less than 0.001), were fertilized when the husband's semen analysis was in the normal range. The proportion of fertilized oocytes was not decreased in cases of polyzoospermia (greater than 130 X 10(6) spermatozoa/ml), but was decreased (P less than 0.05) when the semen analysis revealed other anomalies: oligozoospermia (less than 15 X 10(6) spermatozoa/ml), asthenozoospermia (less than 50% motile cells) or teratozoospermia (greater than 50% abnormal spermatozoa). The proportion of fertilized eggs cleaving in vitro was unaffected by semen quality but was lower when "+" cumulus oocytes were collected than when "++" cumulus oocytes were obtained (58.3% vs. 87.0%, P less than 0.02). In vitro incubation of the oocyte prior to insemination increased the incidence of fertilization by about 28% for both "+" (22.2 to 50.0%) and "++" (65.7 to 93.9%) cumulus oocytes. Finally, 67.6% of "++" cumulus oocytes developed into embryos when the insemination with spermatozoa from normal semen samples was delayed by several hours, compared with only 29.0% when the conditions were suboptimal ("+" cumulus oocyte, abnormal semen analysis or no delay prior to insemination). Eight pregnancies began following the replacement of 38 embryos in 34 patients. Six spontaneous abortions occurred, and chromosomal abnormalities were proven in the two cases analyzed. Two pregnancies continued for more than 3 months, resulting in term deliveries of two normal babies.

Homogeneous Mixing of Honeybee Semen by Centrifugation

Abstract: SummaryDistribution of semen in the spermatheca of the queen honeybee (Apis mellifera comica) was determined by using genetic markers (cd, pe, di). With washing and centrifugation the semen was mixed homogeneously. If the semen was not treated the distribution was not homogeneous, and sperms of each mutant fertilized the eggs laid by the queen at constant frequencies.