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PubMed1994

Preservation of ejaculated and epididymal stallion spermatozoa by cooling and freezing.

The suitability of ejaculated and epididymal stallion spermatozoa for cooled storage (5 degrees C) and cryopreservation was examined in 5 ejaculates from each of 6 stallions and in spermatozoa recovered from the cauda epididymidis after castration of these stallions. The percentage of progressively motile spermatozoa, examined by subjective estimation (cooled samples) or by computerized analysis (frozen-thawed samples), was used as parameter. In ejaculated semen samples containing 5 and 25% seminal plasma in a skim milk glucose extender, the lower amount of seminal plasma supported spermatozoal motility significantly better throughout storage at 5 degrees C. Addition of 5 or 25% seminal plasma to perfused epididymal spermatozoa (0% seminal plasma) resulted in a significant stimulation of spermatozoal motility by 25% seminal plasma at 0 h (P<0.05) and to a lesser extent at 24 and 48 h. Post-thaw motility of ejaculated as well as epididymal spermatozoa was not influenced by slow cooling to 15 degrees or 5 degrees C with or without glycerol prior to rapid freezing in liquid nitrogen vapor. During cooled storage, seminal plasma had a stimulatory effect on epididymal spermatozoa and depressed motility in ejaculated spermatozoa. Results on cryopreservation indicate that freezability of equine spermatozoa is already determined when spermatozoa leave the tail of the epididymis.

Journal Article↗
PubMed1996

Tissue kallikrein activity in seminal plasma of bovine ejaculates with different quality.

Tissue kallikrein activity was monitored in seminal plasma from 3 groups of bovine ejaculates: those with normal total sperm motility (78.43%), with reduced sperm motility (49.29%), and with reduced sperm count (0.68 x 10(9) cells/ml). The tissue kallikrein activity was measured spectrophotometrically by using the specific chromogenic substrate S-2266. It was found that the semen samples with normal sperm motility manifested 1.083 microkat/L, on an average, or 29% higher than the activity recorded in ejaculates with reduced sperm motility (P < 0.05). After storage of a group of ejaculates of normal quality for 5 h at room temperature, sperm motility dropped by approximately 80%, expressed as a percentage of the initial motility, while the tissue kallikrein activity in the respective seminal samples decreased by 23%. No significant differences were found in kallikrein activity between ejaculates with normal and reduced sperm counts. It is concluded that a relationship exists between the level of tissue kallikrein activity in the seminal plasma of bovine ejaculates and sperm motility.

Journal Article↗
PubMed1996

The occurrence and identification of round spermatids in the ejaculate of men with nonobstructive azoospermia.

OBJECTIVE: To evaluate the occurrence of round spermatids in the ejaculate of men suffering from nonobstructive azoospermia as a prerequisite for the therapeutic use of round spermatid injection into oocytes. DESIGN: Prospective study. SETTING: University-based diagnostic and research laboratory. PATIENT(S): Azoospermic men previously thought to be definitively infertile whose cases were revisited in view of the novel method of fertilization by round spermatid injection into oocytes. INTERVENTION(S): Smear preparations of cells from patients' ejaculates were stained by three different methods: Papanicolaou, fluorescein-labeled Pisum sativum agglutinin binding, and antiacrosin antiserum immunolabeling. MAIN OUTCOME MEASURE(S): Search for round spermatids in smear preparations of cells from azoospermic ejaculates. RESULT(S): Of 124 azoospermic men examined, 36 (69%) had round spermatids in the ejaculate. the quantity of round spermatids was not related to serum FSH concentration. CONCLUSION(S): Variable quantities of round spermatids can be detected in the ejaculate of most patients with nonobstructive azoospermia. High serum FSH levels do not predict the absence of spermatids.

Acrosin↗
PubMed1977

Antibody-coated bacteria in the ejaculate: a possible test for prostatitis.

An immunofluorescence technique was used to study antibody coating of bacteria in ejaculates from 14 healthy individuals and 51 patients with complaints compatible with the diagnosis of prostatitis. Quantitative bacteriological cultures in the ejaculate were positive in 2 healthy individuals (14%) and in 25 patients with prostatic symptoms (49%). Antibody-coated bacteria could be demonstrated in 25 patients with prostatic symptoms (49%), 8 of whom had negative bacterial cultures, but in none of the healthy individuals. All 5 patients with epididymitis had antibody-caoted bacteria in the ejaculate. The 13 patients with antibody-coated bacteria in the ejaculate were given antibiotic treatment and the bacteria disappeared in 8 cases. The results document the presence of a (presumably local) immune response in bacterial prostatitis. Antibody coating of bacteria in the ejaculate seems to be helpful in the diagnosis of bacterial prostatitis.

Antibodies↗
PubMed1996

Ejaculation increases scrotal surface temperature in bulls with intact epididymides.

The objective of the study was to determine the effects of ejaculation on scrotal surface temperature (SST) measured with infrared thermography in bulls. In 18 Holstein bulls (18 mo old), sexual stimulation and spontaneous ejaculation (into an artificial vagina) increased SST at the bottom of the scrotum (0.9 degrees C; P < 0.0001). In 11 Angus bulls (1 yr old) electroejaculation increased both bottom and average SST (1.7 degrees C; P < 0.005 and 0.9 degrees C, P < 0.05), while in 12 Simmental cross bulls (2 yr old) electroejaculation significantly increased top, bottom and average SST (1.0, 1.2 and 1.1 degrees C, respectively). However, there was no significant increase in SST following electroejaculation in 15 Simmental cross bulls (2 yr old) with caudal epididectomies. The increase in SST was attributed to a localized increase in SST over the cauda epididymides, perhaps due to heat produced by contraction of the cauda epididymides during ejaculation. The results support the hypothesis that spontaneous ejaculation or electroejaculation increases SST and that this response is mediated by the cauda epididymides. Infrared thermography of the scrotum for evaluation of scrotal/testicular thermorégulation for clinical or research purposes should be performed before semen collection since thermography conducted soon after ejaculation may be misleading.

Journal Article↗
PubMed1967

Lipid composition and metabolism in testicular and ejaculated ram spermatozoa.

1. Spermatozoa collected directly from the testis of the conscious ram contain 25% more phospholipid than ejaculated spermatozoa. The concentration of lecithin, phosphatidylethanolamine and ethanolamine plasmalogen was greater in testicular spermatozoa; little difference was observed in choline plasmalogen. Both types of spermatozoa had significant amounts of cardiolipin and alkyl ether phospholipid. 2. The fatty acids in the phospholipid extracted from testicular spermatozoa have a very high content of palmitic acid. The phospholipids of ejaculated spermatozoa contained less palmitic acid, but more myristic acid. 3. Ejaculated spermatozoa contained less acyl ester and cholesterol. It is suggested that lipids are a source of substrate for spermatozoa during their passage through the epididymis. 4. Testicular spermatozoa when incubated with [U-(14)C]glucose incorporated more radioactivity into the glycerol part of the phospholipid and neutral lipid fractions than did ejaculated cells. The distribution of radioactivity in the individual phospholipids and neutral lipids was similar for both cell types. No radioactivity was detected in choline plasmalogen, which accounted for approx. 40% of the total phospholipid. 5. Testicular spermatozoa incorporated more radioactivity from glucose into formate than into acetate, whereas a higher proportion of radioactivity was found in acetate in ejaculated cells. 6. The implications of these lipid changes in the process of spermatozoal maturation are discussed.

Animals↗
PubMed2000

Inactive free : total prostate specific antigen ratios in ejaculate from men with suspected and known prostate cancer, compared with young control men.

OBJECTIVE: To measure free : total prostate specific antigen (PSA) ratios in ejaculate from men with suspected and known prostate cancer, and in young control men, to determine if this ratio might be useful in discriminating benign from malignant prostatic conditions. Patients, subjects and methods Forty-seven men with prostate cancer (positive biopsies), 52 men with suspected prostate cancer but who had negative biopsies and 28 young men (< 30 years old) and with no family history of cancer, provided either a single ejaculate specimen (total 59) or multiple specimens (total 193) on subsequent occasions. Free and total PSA were measured using appropriate assays. All specimens were diluted in a PSA-negative female serum pool. RESULTS: The median free : total PSA ratios were 0.76-0.81 among the patient groups and control men, and there was no statistical difference between the groups. These data presumably only reflect the inactive component of free PSA, given that any alpha2-macroglobulin or alpha1-antichymotrypsin in the assay serum diluent was likely to have bound the active free PSA component in these samples. Similar results were obtained from those providing single and multiple samples, suggesting that a single specimen is sufficient to reflect the seminal plasma free : total PSA ratio over that period. There was no relationship between seminal plasma free : total PSA ratio and age for the controls or the positive biopsy group, although there was a negative relationship (i.e. a decline with age) that almost reached significance in those with negative biopsies (P = 0.058, R2 = 0.07). CONCLUSIONS: This is the first report of free : total PSA ratios in the ejaculate of men with suspected and known prostate cancer compared with young control men. Although no significant changes were detected in the free : total PSA ratios in ejaculate, these results may be confounded by differences in ratios with age, as is the case for serum PSA or different molecular forms of PSA. Indeed, these data suggest that a large proportion of free PSA in seminal plasma may be inactive. Further studies are needed to determine the potential utility of measuring free : total PSA, or other candidate markers, in ejaculate to better discriminate benign from malignant prostate disease.

Adult↗
PubMed1994

High concentration of soluble interleukin-2 receptors in ejaculate with low sperm motility.

The various cytokines are involved in infection and immunological reactions and thus may be involved in impairment of sperm function. The present study determined the concentrations of interleukin-1, interleukin-6 and soluble interleukin-2 receptors in ejaculates of normal donors and patients suffering from pure asthenozoospermia and patients suffering from oligoteratoasthenozoospermia. Ejaculates of patients attending the fertility clinic in Hadassah Mount Scopus Hospital, Israel, were evaluated. The patients were divided into three groups: (i) controls (n = 20), (ii) pure asthenozoospermia (n = 30), (iii) oligoteratoasthenozoospermia (n = 36). Significantly higher concentrations of soluble interleukin-2 receptors were found in the ejaculates of patients with pure asthenozoospermia 2243.1 +/- 473.6 mu/ml (mean +/- SE) as compared with controls 673.5 +/- 281.3 mu/ml (P < 0.05) and oligoteratoasthenozoospermic patients 1012.3 +/- 206.9 mu/ml (P < 0.05). Interleukin-1 and interleukin-6 concentrations were comparable in all groups studied. Our results show that human ejaculates contain interleukin-1, interleukin-6 and soluble interleukin-2 receptors. High concentrations of soluble interleukin-2 receptors in the ejaculates of asthenozoospermic patients may be related to either infectious or immunological processes.

Humans↗
PubMed2004

Sexual conflict and anti-aphrodisiac titre in a polyandrous butterfly: male ejaculate tailoring and absence of female control.

Males of the green-veined butterfly Pieris napi synthesize and transfer the volatile methyl salicylate (MeS) to females at mating, a substance that is emitted by non-virgin females when courted by males, curtailing courtship and decreasing the likelihood of female re-mating. The volatile is released when females display the 'mate-refusal' posture with spread wings and elevated abdomen, when courted by conspecific males. Here, we assess how the amount of MeS released by courted females changes over time since mating, and whether it is influenced by the frequency with which females display the mate-refusal posture. We also assess whether males tailor the anti-aphrodisiac content of ejaculates with respect to the expected degree of sperm competition, by comparing how males allocate MeS proportionately to first and second ejaculates in relation to ejaculate mass. The results show that females housed for 5 days in individual cages where they were able to fly and oviposit normally, released similar amounts of MeS. However, females housed together for the same period of time, causing them to frequently display the mate-refusal posture, released significantly lower levels of MeS than the individually housed females. This indicates that female display of the mate-refusal posture depletes their anti-aphrodisiac stores, and suggests that females are unable to voluntarily control their release of the anti-aphrodisiac. A comparison of relative proportion of MeS transferred by males in their first and second ejaculates showed that proportionately more MeS was allocated to the first ejaculate, in accordance with the idea that these are tailored to delay female re-mating.

Analysis of Variance↗
PubMed2003

Female life span and fertility are increased by the ejaculates of preferred males.

In animals with internal fertilization, sperm competition among males can favor the evolution of male ejaculate traits that are detrimental to females. Female mating preferences, in contrast, often favor traits in males that are beneficial to females, yet little is known about the effect of these preferences on the evolution of male ejaculates. A necessary condition for female preferences to affect the evolution of male ejaculate characteristics is that females select mates based on a trait correlated with ejaculate quality. Previous work has shown that females of the variable field cricket, Gryllus lineaticeps, prefer males that produce calling songs containing faster and longer chirps. In this study, we tested the hypothesis that females receive more beneficial ejaculates from preferred males. Females were placed on either a high- or a reduced-nutrition diet then mated twice to a male of known song phenotype. Females received only sperm and seminal fluid from males during these matings. There was no effect of male song phenotype on any fitness component for females on the high-nutrition diet. Reduced-nutrition females mated to males that produced preferred song types, however, lived longer, produced more eggs, produced more fertile eggs, and had a higher proportion of their eggs fertilized than those mated to other males. The life-span benefit was positively associated with male chirp duration, and the reproductive benefits were positively associated with male chirp rate. We explored two possible mechanisms for the life span and reproductive benefits. First, a path analysis suggested that part of the effect of male chirp duration on female life span may have been indirect; females mated to males that produced longer chirps showed delayed oviposition, and females that delayed oviposition lived longer. Males that produce longer chirps may thus transfer fewer or less potent oviposition stimulants to females in their seminal fluid. Second, there was a positive correlation between male chirp rate and the number of sperm transferred to females. The fertility benefit may thus have resulted from females receiving more sperm from males that produce faster chirps. Finally, there was a negative phenotypic correlation between male chirp rate and chirp duration, suggesting that females may have to trade off the life span and reproduction benefits when selecting a mate.

Animal Communication↗
PubMed1986

Ejaculate composition after masturbation and coitus in the human male.

The levels of fructose, acid phosphatase, citric acid, zinc, maltase, testosterone, prostaglandin E, putrescine, spermidine and spermine were compared in ejaculates collected by masturbation and after coitus in the same individuals. Coitus was associated with a significantly larger semen volume and increased concentrations and total amounts of prostaglandin E and polyamines in the ejaculate. In contrast, the concentrations of the conventional glandular parameters remained relatively unaffected, although a slight dilution of the seminal vesicle contribution to the ejaculate occurred after coitus. In the majority of cases, ejaculates provided by masturbation were significantly enriched in spermatozoa, but the total contributions under both conditions remained the same. It is concluded that the assessment of semen quality in male individuals using the conventional parameters of accessory gland function can be made with ejaculates provided by masturbation or coitus.

Acid Phosphatase↗
PubMedDate not supplied

Determination of fluconazole concentrations in the prostatic and seminal vesicle fluid (split ejaculate).

The concentrations of fluconazole, a systemic antifungal agent, were determined in the split ejaculate of seven healthy volunteers. Following once-daily oral administration of fluconazole 200 mg over a period of 5 days, the fluconazole levels were determined in ejaculate samples which had been divided into two consecutive fractions upon collection. Simultaneously, a blood sample was drawn for the determination of fluconazole serum levels. It was shown that the fluconazole concentrations determined in both ejaculate fractions corresponded to those found in the serum. As the first fraction of the ejaculate originates in the prostate, the prostatic fluconazole concentrations are likely to be similar to the fluconazole levels found in th ejaculate.

AIDS-Related Opportunistic Infections↗
PubMed2006

Motility and plasma membrane integrity of spermatozoa in fractionated stallion ejaculates after storage.

With the aim of investigating properties of stallion seminal plasma to eventually improve semen-handling techniques, sperm motility and plasma membrane integrity were analysed in different fractions of the ejaculates after storage. Semen was collected using a computer-controlled automated phantom that separates the ejaculates into five successive cups. Samples containing seminal plasma and skim milk extender were compared with samples stored in skim milk extender after the removal of seminal plasma by centrifugation. Fractionated ejaculates were stored cooled for 24 h after dilution with extender (Expt 1) or frozen in liquid nitrogen (Expt 2). In Expt 1, cup 1 was pre-sperm fluid, cups 2 and 3 sperm-rich fractions, and cup 4 sperm-poor fractions. In Expt 2, cups 1 and 2 were sperm-rich fractions, and cups 3 and 4 sperm-poor fractions. One sample (WE) represented the whole ejaculate in both experiments. Motility parameters were determined with a Hamilton-Thorn Motility Analyzer, and plasma membrane integrity was assessed using carboxyfluorescein diacetate and propidium iodide staining and fluorescence microscopy. The removal of seminal plasma lowered motility values, but not plasma membrane integrity, in both experiments. No significant differences between cups were observed after cooled storage. The cups differed significantly in most post-thaw motility parameters, and the sperm-rich fraction showed higher post-thaw motility than the whole ejaculate.

Animals↗
PubMed1993

Lack of expression of HLA antigens on immature germ cells from ejaculates with antisperm antibodies.

PROBLEM: The lack of expression of HLA antigen on immature germ cells from ejaculates with antisperm antibodies has been reported. METHOD: The expression of human leukocyte antigens on immature germ cells from ejaculates with antisperm antibodies (ASA) was investigated by indirect immunofluorescence using a panel of monoclonal antibodies (MAb) and automated flow cytometry. Patients were divided into two groups: fertile (prevasectomic; N = 10), and ejaculates with ASA (10 samples with IgG and IgA ASA, and five semen samples with only IgG ASA). ASA were detected on sperm using the direct immunobead test. After centrifuging semen samples on a Ficoll-Hypaque gradient, round cells obtained at the gradient interface were gated by a flow cytometer. The "immature germ cell window" was defined in terms of cellular volume and granularity. RESULTS: The percentage of gated round cells from semen samples that reacted with anti-CD45 was always less than 5%, and with anti-CD44 less than 3%. This lack of reactivity of gated round cells with MAb specific for leukocytes and epithelial cells suggests that they were immature germ cells. Immature germ cells were unreactive with W6/32 and anti-beta-2-microglobulin MAb, which suggests that these cells do not express HLA class I molecules. Similarly, no reactivity of the immature germ cells with the MAb that recognize HLA class II molecules was found. No significant differences were observed in the expression of HLA molecules on immature germ cells between the different semen samples studied: fertile, and ejaculates with ASA. CONCLUSION: The presence of ASA in ejaculate is not associated with abnormal HLA antigen expression on immature germ cells.

Animals↗
PubMed1984

A review of female ejaculation and the Grafenberg spot.

A review of recent works related to the concept of female ejaculation, defined as a partial, infertile homologue to male ejaculation, indicated that we have insufficient evidence to accept or reject the concept on scientific grounds. The claim that "female ejaculate" is similar to semen from a vasectomized male is without foundation. Although recent discussion of female ejaculation has tended to focus on whether or not the fluid sometimes expelled from the urethra upon orgasm is invariably urine, the Bartholin's glands are another potential source of orgasmic expulsions which could properly be called "female ejaculate." "Grafenberg spot" is a neologism referring to an area which some women report is erotically sensitive when massaged via the anterior vaginal wall. "Urethral sponge" is another term which seems to refer to this anatomical/phenomenological reality. The relationship between stimulation of this area and subsequent orgasmic expulsion has yet to be studied systematically.

Acid Phosphatase↗
PubMed1996

Chromatin defects in normal and malformed human ejaculated and epididymal spermatozoa: a cytochemical ultrastructural study.

Cytochemical defects in chromatin were examined by transmission electron microscopy (TEM) after the staining by alcoholic phosphotungstic acid (PTA) of normal and malformed ejaculated spermatozoa from 35 male partners of infertile couples, and in six sperm samples retrieved from the caput epididymidis of men affected by obstructive azoospermia. PTA staining was also analysed in normal ejaculates of fertile men after incubation of the washed spermatozoa with dithiothreitol (DTT) to reduce disulfides to thiols, or with DTT followed by iodoacetamide, a blocking agent for thiol groups. PTA stained 63 (27-100)% of malformed heads and 25 (10-100)% of normal sperm heads (median (range) n = 35; P = 0.0001, Wilcoxon matched pairs test). The percentage of normal heads stained by PTA was negatively correlated with the percentage of heads of normal form, with condensed chromatin and a normal acrosome (Spearman r = 0.75; P = 0.0001), and positively correlated with the percentage of malformed heads after conventional TEM analysis (Spearman r 0.60; P = 0.0001). Staining with PTA in normal heads was not correlated with the presence of non-condensed chromatin in otherwise normal sperm heads evaluated by conventional TEM analysis. In spermatozoa recovered from the caput epididymidis, 15% of normal heads were stained with PTA, significantly fewer than in ejaculated sperm samples (P = 0.014). The reduction of disulfides to thiols was associated with PTA staining of all normal heads, and this was prevented by incubation with iodoacetamide. We conclude that PTA staining of the nuclei of human ejaculated spermatozoa may indicate a defect of chromatin condensation, owing to an excess of free thiol groups. The lower percentage of normal epididymal sperm heads that stained with PTA in cases of obstructive azoospermia compared with ejaculated sperm may be related to an overoxidation of thils owing to the ageing of spermatozoa.

Chromatin↗
PubMed1987

Phosphate, zinc, calcium, citric acid, and acid phosphatase in human ejaculates as related to coagulation/liquefaction.

Variations in the coagulation-liquefaction property of human seminal coagula have rarely been correlated to their ingredients. A number of seminal components were evaluated in relation to the degree of coagulation and the liquefaction time of human ejaculates. Correlation analyses suggested that the vesicular coagulatory activity can better be assessed by the phosphate level rather than the fructose and N-acetylamino sugar levels. The results showed that the fresh ejaculates with a smaller amount of coagulum possess significantly higher prostatic but lower vesicular activities in comparison with that of the better coagulating samples. The study also revealed that liquefaction time-based fractionation of the whole ejaculate seems to give an idea about the origin of some seminal components like split ejaculation technique. It was concluded that the material characteristics of human ejaculate can be approximated simply by determining its degree of coagulation or liquefaction time.

Acid Phosphatase↗
PubMedDate not supplied

Morphologically distinct sperm subpopulations defined by Fourier shape descriptors in fresh ejaculates correlate with variation in boar semen quality following cryopreservation.

This study investigated two hypotheses: 1) that consistent between-boar variation in frozen semen quality exists and is genetically determined, and 2) that morphologically distinct subpopulations of spermatozoa exist within fresh boar ejaculates and that the incidence of these subpopulations is correlated with semen quality following cryopreservation. Five ejaculates were collected from each of 15 boars (5 boars from each of 3 breeds). An objective sperm morphology analyzer used Fourier shape descriptors to describe variation in the morphology of 300 spermatozoa per ejaculate before freezing. Semen was diluted into a commercial freezing buffer (700 mOsm/kg, 3% glycerol) and 5 straws (0.5 mL) per ejaculate were cryopreserved (to -5 degrees C at 6 degrees C/min, then -5 degrees C to -80 degrees C at 40 degrees C/min). Semen was assessed for percentage of motile cells and motility characteristics (with computer-aided sperm analysis), plasma membrane integrity (SYBR-14 positive), and acrosome integrity (fluorescein-labeled peanut agglutinin positive). Consistent between-boar variability was detected for post-thaw sperm motility (P < .01), membrane integrity (P < .01), acrosome integrity (P < .01), curvilinear velocity (P < .01), straight-line velocity (P < .05), beat cross-frequency (P < .05), and amplitude of lateral head displacement (P < .01). Three morphologically distinct subpopulations of spermatozoa, defined by Fourier descriptors, were detected. The proportion of these subpopulations within the fresh ejaculate correlated with semen quality assessments made following cryopreservation. These findings support the hypothesis that consistent interindividual variation in sperm freezability is genetically determined and may relate to processes that occur during spermatogenesis. Subsequent characterization of these genetic differences between "good" and "poor" freezers may ultimately identify biophysical components of the spermatozoa that are essential for successful cryopreservation.

Acrosome↗