Concentration of prostaglandins E and F,fructose and glycerylphosphorylcholine in ram semen obtained by electro-ejaculation or artificial vagina and in vesicular fluid

The concentration of spermatozoa in electrically ejaculated ram semen was lower than in semen obtained by an artificial vagina. Glycerylphosphorylcholine concentrations were also lower in the electrically ejaculated semen and there was a high correlation between sperm and glycerylphosphorylcholine concentrations.Seminal fructose, prostaglandin E (PGE) and prostaglandin F (PGF) levels did not differ significantly between the two methods of collection but there was greater variability between rams when they were electrically ejaculated.The concentration of fructose in the vesicular secretion of rams was less variable and higher than in seminal plasma whereas PGE or PGF concentrations were not significantly different in the two fluids.     

The effect of semen extender,seminal plasma and raw semen on uterine and ovarian blood flow in mares

Transrectal color Doppler sonography was used to evaluate the effect of intrauterine infusion of skim milk semen extender, seminal plasma and raw semen on the endometrium and blood flow in the uterine and ovarian arteries in mares. Six Trotter mares (mean age: 12 years) were examined during estrus in three cycles. Each mare received an intrauterine infusion of 20 ml of skim milk semen extender, seminal plasma or raw semen during estrus in one of three cycles. Blood flow measurements in both uterine and ovarian arteries and the determination of intrauterine fluid via sonography were performed before each infusion and 1, 3, 6, 12, and 24 h after infusion. Forty-eight hours later, the intrauterine infusion and measurements were repeated using the same time intervals. Changes in blood flow were detected using transrectal color Doppler sonography and were evaluated using the mean time-averaged maximum velocity (TAMV) of the blood flow. Cytological and bacteriological examination of uterine swabs performed 48 h after the second infusion revealed less inflammation and bacterial growth in mares infused with skim milk semen extender than in those infused with seminal plasma or raw semen. There was an increase in intrauterine fluid as early as 1 h after infusion of any of the substances. The infusion of skim milk semen extender had no effect on uterine blood flow. Within 1 h after infusion of seminal plasma or raw semen, there was an increase in the TAMV values of both uterine arteries (P<0.05). In contrast, ovarian blood flow increased only in the artery ipsilateral to the preovulatory follicle and only after the infusion of raw semen (P<0.05). In conclusion, the changes in uterine perfusion observed after intrauterine infusion may be associated with endometrial inflammation and vasodilatory components in the seminal plasma, whereas the changes seen in ovarian blood flow are possibly attributable to the interaction between sperm and oviduct.     

Seasonal thyroidal activity and reproductive characteristics of Iranian fat-tailed rams

An experiment was conducted to study seasonal variations of thyroidal activity, serum testosterone concentration and seminal characteristics of two breeds of Iranian fat-tailed sheep. Eight 3 to 4-year-old rams of Ghezel and Mehraban breeds (4 rams/breed) were randomly selected from a flock of fertile rams. Semen was collected by using an artificial vagina, and blood samples were obtained via jugular vein. Seminal characteristics, scrotal length, width, and circumference, and blood parameters were measured at 3-week intervals. Time of the year significantly (P < 0.05) affected the volume of semen, sperm concentration, percent live sperm, percent normal sperm, total sperm number, seminal pH, seminal lactate dehydrogenase (LDH) concentration, scrotal circumference, scrotal width, and serum levels of testosterone, cholesterol, triiodothyronine (T(3)), tetraiodothyronine (T(4)), free T(4) index (FT(4)I), and TSH. Scrotal length was not affected by the time of sampling (P > 0.05). The effect of breed on the serum concentrations of hormones and cholesterol was not significant. Volume of semen, percent normal sperm, and total number of normal sperm were significantly (P < 0.05) higher in Mehraban but scrotal circumference, scrotal width and scrotal length were greater in Ghezel rams. Breedxtime interaction effect was significant for T(4), FT(4)I, sperm concentration, and total sperm number. The largest values for TSH, T(4), FT(4)I, testosterone, total sperm number, percent normal sperm, percent live sperm, sperm concentration, volume of semen and scrotal circumference were found from early summer to winter and the lowest values were recorded at the end of spring and in early summer. In both breeds, high and negative correlations (r = -0.7 to -0.8) were found between LDH levels in semen and percent live sperm, percent normal sperm and total sperm number (P < 0.01). It is unlikely that such variations in seminal quality would affect the fertility under natural mating.     

Impact of seminal and serum zinc on semen quality and hormonal status: A population-based cohort study of Russian young men

BackgroundTrace elements are important factors in human reproductive health. Among them, special attention is paid to zinc, which is an essential trace element and is necessary for the normal functioning of the male reproductive system and the process of spermatogenesis. The aim of the study was to investigate the association between seminal and serum zinc concentrations and semen quality and reproductive hormone levels in population of Russian young men.MethodsThe study population consisted of 626 young Russian men (median age 22.5 years), recruited from the general population, regardless of their fertility status. Each participant provided semen and blood sample, information about his lifestyle and ethnicity. Semen quality (sperm concentration, motility and morphology), reproductive hormone levels (testosterone, estradiol, LH, FSH and inhibin B), and serum and seminal zinc concentrations were evaluated. The semen samples were analyzed according to the WHO laboratory manual (WHO, 2010). Serum hormones were measured by enzyme immunoassay, zinc concentrations were determined using spectrophotometry and direct colorimetry without deproteinization.ResultsZinc was present in the seminal plasma in a significantly higher concentration than in the blood serum (median serum Zn concentration was 23.6 μmol/L vs seminal Zn concentration 1571.8 μmol/L). The seminal zinc concentration was positively related to the total sperm count, sperm concentration, progressive motility and normal morphology (Spearman’s test: 0.221; 0.286; 0.269; 0.183, respectively, p < 0.001), while the serum Zn concentration was negatively related to serum testosterone and estradiol levels (r = −0.249 and r = −0.096, respectively, p < 0.001−0.05). It was found that the seminal Zn content in men with normal semen quality was higher compared to men with lowered semen quality (means: 6.37 and 5.03 μmol/ejaculate, respectively, p < 0.001). Similarly, the semen volume, total sperm count, sperm concentration, progressive motility, normal morphology and the serum testosterone level in men with the seminal Zn deficiency were lower than in men with the normal seminal Zn content.ConclusionBased on the results of our population-based study, seminal Zn levels were closely associated with semen parameters in young men, so Zn deficiency may be an important risk factor for lowered semen quality. Seminal Zn determinations should be considered as a useful tool in addition to other parameters in assessing male fertility.     

Trace elements and lipid peroxidation in human seminal plasma

In the present study, the concentrations of copper, iron, zinc, and malondialdehyde in human seminal plasma were measured and correlated with the sperm count and motility in human semen. Copper, iron, and zinc were analyzed by atomic absorption spectrometry, whereas malondialdehyde was measured by high-performance liquid chromatography. The malondialdehyde concentrations in asthenospermia and oligoasthenospermia were significantly higher than in normospermia. Copper and iron levels were higher in asthenospermia, whereas the zinc concentrations in both oligospermia and asthenospermia were lower than in normal controls. A negative correlation (r = -0.28, p < 0.05) between the malondialdehyde concentration and sperm motility was observed in the abnormal groups. There was no association among copper, iron, zinc, and malondialdehyde in seminal plasma. We concluded that changes in trace elements may be related to sperm quality and that lipid peroxidation, although it is not promoted in the seminal plasma by copper or iron or ameliorated by zinc, may be involved in the loss of sperm motility.     

ATP contents of human semen, seminal plasma and isolated sperm at time intervals after ejaculation

1. ATP was estimated in 105 samples of human semen, seminal plasma and sperm of normozoospermic and oligozoospermic origins at time intervals after ejaculation. 2. In semen with sperm counts up to 40 millions per ml and in seminal plasma, ATP levels were lower than in specimens with higher sperm counts. 3. In isolated sperm, the ATP content decreased with the increase in sperm density. 4. The decrease in ATP after 24 hr was the highest in sperm, lower in semen and the lowest in seminal plasma, being maximal in specimens with high sperm counts.     

Reproductive characteristics of Rayini male goats of Kerman province in Iran

An artificial vagina was used to collect semen from 14 native Iranian Rayini goats, at 15-day intervals starting on 1 July and ending on 30 December 2000. Testicular size, semen volume, sperm concentration, percent live sperm, percent normal sperm, and total number of live-normal sperm were significantly higher during the summer months. The average semen volume, percent live sperm and percent abnormal sperm during the sampling period varied between 1 and 1.4 mL, 60 and 78%, and 7 and 13%, respectively. The total number of live and normal sperm in the ejaculate during the sampling period varied from 1000 to 2500 million. Testicular size, semen volume and the total number of live and normal sperm were significantly greater in bucks weighing 55-60 kg as compared with 50-54 kg. Seminal fluid pH values were significantly lower from July to October (pH <6) than the values from November to December (pH >6.1). Lowest level of lactate dehydrogenase in the seminal fluid was recorded in early September (2.2 U/mL) and the highest level in November (2.5 U/mL). Seminal fluid K ion level increased gradually from July (52 mg/dL) to the November (97 mg/dL). Variation of seminal fluid Na ion concentration (71-74 mg/dL) was not significant during the sampling period. The correlation coefficients of total number of live-normal sperm with seminal fluid K level (r=-0.65) and LDH (r=-0.36) were negative (P<0.01). The data indicated that the semen quality and quantity of Rayini bucks were higher during summer and early autumn.     

The associations among semen quality,oxidative DNA damage in human spermatozoa and concentrations of cadmium,lead and selenium in seminal plasma

To explore the associations among semen quality, oxidative DNA damage in human spermatozoa and concentrations of cadmium, lead and selenium in seminal plasma, 56 non-smoking subjects were asked to collect semen by masturbation into a sterile wide-mouth metal-free plastic container after 3 days of abstinence. The conventional semen parameters were analysed. The concentrations of Cd, Pb and Se in seminal plasma were detected using atomic absorption spectrophotometer. 8-OHdG levels in sperm DNA were measured using HPLC-EC. The results showed that the geometric mean concentrations of Cd, Pb and Se were 0.78, 7.8 and 51.4 microg/l, respectively. The geometric mean of 8-OHdG/10(6) dG was 51.4 (95% CI: 21.5-123.0). A significant inverse correlation exists between Cd and sperm density (r=-0.28, P<0.05), and between Cd and sperm number per ejaculum (r=-0.27, P<0.05). In contrast, there was a significantly positive correlation between Se and sperm density (r=0.50, P<0.01), between Se and sperm number (r=0.49, P<0.01), between Se and sperm motility (r=0.40, P<0.01), and between Se and sperm viability (r=0.38, P<0.01). No statistically significant correlation was observed between Pb and semen quality. A significant inverse correlation was observed between 8-OHdG and sperm density (r=-0.34, P<0.01), between 8-OHdG and sperm number per ejaculum (r=-0.30, P<0.01), and 8-OHdG and sperm viability (r=-0.24, P<0.05). 8-OHdG was significantly correlated with Cd in seminal plasma (r=0.55, P<0.01). A significant but weak positive correlation was found between 8-OHdG and Pb concentration in seminal plasma (r=0.28, P<0.05). In contract, a significant inverse correlation was observed between 8-OHdG and Se concentration in seminal plasma (r=-0.40, P<0.01). The results indicate that Cd in seminal plasma could affect semen quality and oxidative DNA damage in human spermatozoa. Se could protect against oxidative DNA damage in human sperm cells. Pb did not appear to have any association with the semen quality when concentration of Pb in seminal plasma was below 10 microg/l.     

Estrogenic octylphenol affects seminal fluid production and its biochemical composition of eelpout (Zoarces viviparus)

Estrogenic chemicals such as alkylphenols (APs) have been shown to disrupt the reproductive system of male fish. In the present study, the effects of the estrogenic chemical octylphenol (100 microg g(-1)) and 17 beta-estradiol on the semen production and the biochemical composition of the seminal fluid of the viviparous eelpout (Zoarces viviparus) were investigated at the time of spawning. After 10 days of octylphenol or estrogen treatment, vitellogenin (Vtg) synthesis was induced as indicated by increased plasma vitellogenin concentration. In accordance with the increased vitellogenin concentration, hepatosomatic index (HSI), total protein concentration, and total calcium concentration were also increased, and free amino acids concentration was decreased in blood plasma. Octylphenol treatment caused a decrease in the gonadosomatic index (GSI) and the milt volume and an increase in the spermatocrit. The histological examination revealed that octylphenol affected the normal lobular structure, including the Sertoli cells. In the majority of the octylphenol-treated fish, trapped sperm cells were observed in parts of the seminiferous lobules and the sperm ducts. The biochemical composition of the seminal fluid was also affected by the octylphenol or estrogen. The seminal plasma concentrations of magnesium, calcium, and total protein were elevated, and the concentration of free amino acids was reduced in the treated fish. This study indicates that octylphenol inhibits the seminal fluid production and changes its biochemical composition in eelpouts.     

Seminal plasma addition attenuates the dilution effect in bovine sperm

Dilution of semen to low cell numbers/dose can result in a bull-dependent reduction in the post-thaw viability of cryopreserved bovine spermatozoa. It is possible that essential seminal plasma components are lacking at the greater dilution rates, thereby contributing to the deleterious effects of semen dilution. Ejaculates of 6 Holstein bulls were diluted to 120 x 10(6) sperm/mL in an egg yolk citrate extender (EYC). Split samples were further diluted to 80, 40, 20 and 4 x 10(6) sperm/mL in EYC extender with (+SP) and without (-SP) the addition of frozen/thawed seminal plasma previously obtained from a vasectomized bull. Serial dilutions for the +SP treatments were calculated and performed such that each dilution contained a volume of seminal plasma equal to the original 120 x 10(6) sperm/mL dilution. Samples were then loaded into 0.5-mL French straws yielding final sperm concentrations of 30, 20, 10, 5 and 1 x 10(6)/dose. Straws from each dilution were analyzed using 2 stain combinations: the sperm viability stain, SYBR-14 and propidium iodide (PI); or the mitochondrial-specific, membrane potential-dependent stain JC-1 along with PI. Split-plot analysis of variance indicated that within bulls, there were greater proportions of viable spermatozoa in aliquots containing added seminal plasma than in aliquots without added seminal plasma (P < 0.05). Contrast analyses showed that sperm viability significantly decreased as sperm concentration decreased in the -SP samples. Although the dilution effect was also observed in the +SP samples, the magnitude of the effect was less than in the -SP samples. At most sperm concentrations, the proportions of spermatozoa that stained with JC-1 were correlated (r > 0.84; P < 0.05) with the percentages of SYBR- 14 stained spermatozoa. Furthermore, the proportions of JC-1-stained spermatozoa were greater in the +SP aliquots than in the -SP samples at a concentration of 10 x 10(6) sperm/0.5 mL. These results suggest that the addition of seminal plasma can be beneficial to sperm viability when semen is diluted to low cell numbers/dose.     

Studies of chemical components of Angora goat seminal plasma

Ejaculates were collected by artificial vagina from 11 Angora goats, once or twice weekly, between April and July in two successive years. The mean +/- SEM ejaculate volumes each year were 0.8 +/- 0.30 and 0.98 +/- 0.52 ml; the sperm concentrations were 3.33 +/- 0.49 and 2.94 +/- 0.45 x 10(9)/ml, and the pH values were 7.01 +/- 0.34 and 7.20 +/- 0.17. The concentrations (mg/100ml) of fructose (875 +/- 97) and lactic acid (73 +/- 17) in goat seminal plasma were sufficiently high to be important substrates for maintenance of sperm motility. Only trace amounts of glucose were present in seminal plasma. The glycerylphosphorylcholine (GPC) concentration of seminal plasma (809 +/- 154 mg 100 ml ) was correlated with whole semen sperm concentration (P < 0.001), indicating that GPC is of epididymal origin. Goat sperm are not likely to utilize GPC as a substrate and its metabolizable derivatives, glycerophosphate (3.3 +/- 1.1 mg 100 ml ) and glycerol (1.8 +/- 1.0 mg 100 ml ), were not present in sufficiently high concentrations to be significant as energy sources for the sperm. The mean concentration of citric acid was 331 mg 100 ml seminal plasma. Colored semen was consistently produced by eight bucks, and in yellow, light yellow and white ejaculates, the seminal plasma riboflavin (mug/ml) concentrations were 5.38 +/- 2.89, 3.09 +/- 0.85 and 1.73 +/- 0.88, respectively. This suggests that the color is due to riboflavin, which is probably produced by the vesicular glands since the concentration of riboflavin in the seminal plasma was correlated with fructose and citric acid levels.     

Influence of seminal plasma on the fecundity of chicken spermatozoa

This study was undertaken to investigate the influence of seminal plasma on the fecundity of chicken sperm. Sperm diluted with either incubated seminal plasma (5 or 37 degrees C for 24 h) or seminal plasma from incubated whole semen (5 or 37 degrees C for 24 h) had lower fertility levels and motility scores than sperm diluted in either fresh seminal plasma or a synthetic diluent. The number of sperm with damaged membranes increased with seminal plasma derived from 37 degrees C incubation. The depressive effect of incubated seminal plasma on semen fertility was eliminated by microfiltering .(0.22 mum) the seminal plasma either before or after incubation. Filtration of seminal plasma was only effective in eliminating the depressive effect on sperm motility when filtering was done after incubation. Filtration of seminal plasma reduced the percentage of damaged sperm in all treatments. It can be concluded that there are factors in seminal plasma that are deleterious to the fecundity of chicken spermatozoa and they may be derived from degenerating sperm and/or various fluids, cells and debris collected with the semen during manual semen collection.     

The concentration of pepsinogen C in human semen and the physiological activation of zymogen in the vagina

The relationship between male infertility and the pepsinogen C content in semen has been investigated. The activation of the seminal pepsinogen C in the vagina has been studied under physiological conditions. Samples of semen from 48 vasectomized males and from 46 males of infertile couples were analyzed for pepsinogen C by radioimmunoassay. No correlation was found between the level of pepsinogen C and seminal characteristics, including sperm concentration, motility, and morphologic features. The mean concentration of pepsinogen C was 42.2 micrograms/ml; the first, second, and third quartile were 18.4, 29.6, and 57.6 micrograms/ml, respectively. No significant difference in the level of pepsinogen C was observed between semen of normal quality, semen of reduced quality, and semen with aspermia. Activation of pepsinogen C occurred within 3 h when semen was incubated at pH below 5.0 at 37 degrees C. Intravaginal activation was investigated in six experiments in which semen from two males was instilled in three females. In four experiments with two couples, post-coital activation was investigated. Pepsin C activity in vaginal fluid was detected an average of 3 h (range 2-5 h) and 5 h (4-7 h) after instillation or ejaculation, respectively. Vaginal pH had then been below 4.5 for approximately 1 h. Pepsin C activity was present in the vagina for more than 24 h thereafter. It is most likely that seminal pepsin C is without influence on the fertilizing spermatozoon. However, pepsin C may exert a local effect in the vagina by degrading seminal proteins, thus preventing an immunogenic response in females.     

Effect of semen collection method on pre- and post-thaw Guirra ram spermatozoa

In this study, we evaluated the potential effect of the method of recovery (artificial vagina or electroejaculation) on the production and quality of Guirra ram spermatozoa cryopreserved for the possible constitution of a sperm bank. In order to address this question, we evaluated the effect of semen collection method on fresh semen quality parameters, including: volume, concentration, production, microscopic analysis (abnormal sperm and intact apical ridge) and sperm motility parameters determined by CASA system. For frozen-thawed semen, we evaluated motility parameters by CASA and intact apical ridge, acrosomal status, assessed by dual staining by IP and FITC-PNA and capacitation status, assessed by M540 and Yo-pro1, using flow cytometry. The main findings from this study were: (i) that electroejaculation resulted in a lower recovery efficiency (80% of the cases), as a consequence of contamination with urine or lack of response to the electrical stimulation; (ii) the fresh seminal quality was not significantly different between recovery methods, except for the concentration of spermatozoa, but total number of spermatozoa and the consequent number of possible seminal doses for artificial insemination were similar; and, (iii) a higher number of stable and functional spermatozoa (higher number of live non-capacitated cells, higher live acrosome intact cells and live acrosome reacted cells) were found for frozen-thawed spermatozoa collected by electro ejaculation than by artificial vagina. According to our results, we are able to develop both methodologies in the creation of the Guirra sperm bank. Assuming the advantages and limitations of both methodologies, in Guirra breed, would enable the rapid constitution of a sperm bank including samples from a large number of non-trained rams in a short period of time, which will increase the genetic variability, and so guarantee the conservation of this breed.     

Species-specific interaction of seminal plasma on sperm–neutrophil binding

Bovine semen is naturally deposited in the vagina and spermatozoa migrate through the cervix into the uterus leaving the bulk of seminal plasma (SP) behind. In equine, both spermatozoa and SP are deposited directly in the uterus and SP reduces sperm binding to neutrophils and prevents the formation of DNA-based neutrophil extracellular traps (NETs). We investigated the role of bovine SP on sperm–neutrophil binding using the four most common bovine semen extenders. Contrary to equine, bovine spermatozoa removed from SP had low binding to neutrophils for up to 3 h, but as little as 10% SP increased sperm–neutrophil binding and NETs formation over time. Similar results were obtained with neutrophils isolated from peripheral blood or from the uterus. Scanning electron microscopy showed that the binding can be mediated by NETs or by direct attachment of the cell membranes for both species. The increased binding with SP reduced the number of free spermatozoa indicating that sperm transport to the site of fertilization (and thus fertility) may be hindered. Surprisingly, egg yolk negated the role of bovine SP on sperm–neutrophil binding compared to all the other semen extenders, but did not alter equine sperm binding to neutrophils. Current artificial insemination in bovine relies heavily on egg yolk extender and introduces variable amounts of SP into the uterus, which naturally remains in the vagina. Our results indicate a need to re-evaluate the composition of semen extenders and the semen processing procedures in relation to sperm transport, longevity and fertilizing ability.     

Variation in lipid profiles within semen compartments—the bovine model of aging

Semen lipid composition was examined in young and mature bulls. Given the specific roles of various semen compartments (i.e., seminal fluid, sperm head, and sperm tail) during fertilization, we hypothesized that altered fatty acid and cholesterol composition of a specific compartment might impair semen quality and sperm function. Semen samples were collected from five mature and five young Holstein Friesian bulls during the winter (December–January). Semen was evaluated by computerized sperm-quality analyzer for bulls and was centrifuged to separate the sperm from the seminal fluid. The sperm fraction was sonicated to separate its head and tail compartments. Cold extraction of lipids was performed, and fatty acids and cholesterol were identified and quantified by gas chromatography. Semen physiological features (concentration, motility, and progressive motility) did not differ between mature and young bulls. However, lipid composition within fractions varied between groups, with prominent impairments in the head compartment. In particular, the proportions of polyunsaturated fatty acids, omega-3 fatty acids, and docosahexaenoic acid in the intact sperm; seminal fluid; and sperm head were lower in semen collected from mature bulls than in that from young bulls. The finding suggests an age-differential absorption and/or metabolism through spermatogenesis. Reduced proportions of major fatty acids in mature bulls might reduce membrane fluidity, which in turn might affect the ability to undergo cryopreservation and/or oocyte-sperm fusion through fertilization.     

The effect of seminal sperm concentration on sperm transport in the reproductive tract of female rabbits treated with progesterone or diethylstilbestrol

Forty-two mature Baladi female rabbits were used in a randomized 3x2 factorial experiment to determine the effects of three treatments (control, progesterone injection: 2 mg/doe and DES injection: 0.1 mg/doe) and two semen sperm cell concentrations (1x10(6) and 60x10(6) sperm/0.25 ml semen on sperm transport and distribution in the female reproductive tract. The injections were given for three consecutive days after which rabbits were injected with 5 IU HCG and inseminated with 0.25 ml semen. The does were sacrificed 10 hrs after insemination and the sperm were recovered and counted from the oviducts, uterine horns, cervices and vagina. Total spermatozoa recovered was high when rabbits were inseminated with 60x10(6) sperm as compared to those inseminated with 1x10(6) sperm. When rabbits were injected with progesterone or DES, the number of sperm recovered relative to the total number of sperm inseminated was high in rabbits inseminated with 1x10(6) sperm, in comparison to those inseminated with 6x10(6) sperm. The number of sperm recovered was highest from cervix which was followed by vagina, uterus and oviducts. DES increased the number of the total sperm recovered while progesterone decreased the number as compared to control. This trend was also observed within the different segments of the reproductive tract and with groups inseminated with 1x10(6) or 60x10(6) sperm/0.25 ml semen. The effect of DES was more obvious with does inseminated with low sperm numbers. Significant correlation coefficients were found between the sperm numbers recovered in the uterus and oviducts and in the cervix and uterus of all groups of rabbits.     

SDS-polyacrylamide gel electrophoresis of buffalo bulls seminal plasma proteins and their relation with semen freezability

The objective of this study was to evaluate the protein profiles of seminal plasma in buffalo bulls and to examine their correlation with semen characteristics. Semen of 10 buffalo bulls were collected by a bovine artificial vagina. Semen characteristics (motility, morphology, viability and concentration) were recorded. A part of the semen sample (1 ml) was diluted by tris-egg yolk-glycerol extender, packed in French straws and was frozen in liquid nitrogen. The straws were later thawed and semen characteristics were compared with those of the fresh semen. Seminal plasma was harvested by centrifugation; treated with cold ethanol and then, underwent SDS-polyacrylamide gel electrophoresis (PAGE). Twenty five protein bands were identified on the gel, of which those of <35.5 kDa were prominent (72% of the bands). Of these protein fractions, 24.5 kDa was significantly correlated with sperm progressive motility in fresh and viability in frozen-thawed semen while 45 kDa bands were correlated with abnormal morphology in frozen-thawed semen; 55 kDa protein fractions were correlated with sperm viability of fresh semen. Progressive motility, viability and abnormal sperm morphology of frozen-thawed semen were highly correlated with these parameters in the fresh semen. In conclusion, seminal plasma protein fractions in buffalo bulls are similar to those reported in other animal species and have some correlations with semen characteristics before and after freezing.     

Biophysical and biochemical characteristics of bactrian camel semen collected by artificial vagina

Seminal characteristics were investigated in Bactrian camel in this study. Semen samples from ten mature Bactrian camel bulls were collected using a modified bovine artificial vagina. The biophysical parameters including volume, color, sperm concentration and fast forward progressive motility, percentage of live sperm and the biochemical parameters including osmolarity, pH, glucose, calcium, phosphorus, chloride, triglycerides, phospholipids, total protein, albumin and non-protein nitrogen concentrations in seminal plasma were measured. The mean time for semen collection was 5.3 +/- 0.29 min. The volume of semen varies from 1.2 to 26 (8.2 +/- 0.7 mls). The majority of semen samples (83.6%) were milky in color and consistency. The average osmolarity of semen was 316.1 +/- 1.48 mOsm/kg H(2)O. The pH of semen was slightly alkaline (7.4 +/- 0.03). The mean concentration of spermatozoa was 414.8 +/- 25.04 x 10(6)cells/ml. The fast forward progressive motility of spermatozoa was 62.4 +/- 1.57%. The percentage of live spermatozoa was 85.6 +/- 1.15. Seminal plasma concentration of glucose was 35.8 +/- 0.9 mg/dl. Non-protein nitrogen, total protein and albumin were 32.5 +/- 2.5, 2200 +/- 100 and 1100 +/- 100mg/dl, respectively. The average concentrations of phospholipids and triglycerides in seminal plasma were 36.4 +/- 2.1 and 101.6 +/- 5.5mg/dl, respectively. The concentrations of calcium, phosphorus and chloride were 8.2 +/- 0.1, 2.9 +/- 1.7 mg/dl and 97.9 +/- 2.9 mEq./l, respectively.     

Dimorphic Sperm Influence Semen Distribution in a Non-copulatory Sculpin Hemilepidotus Gilberti

We used an artificial semen emission test to examine the semen transporting role of parasperm (unflagellated sperm), which are produced along with eusperm (normal sperm) by an incomplete meiosis in the marine sculpin Hemilepidotus gilberti. After separation of contents of semen (eusperm, parasperm, and seminal plasma) by centrifugation, the distance traveled by semen discharged vertically from a syringe into seawater was compared among various test semen; eusperm semen (eusperm re-mixed with seminal plasma), parasperm semen (parasperm re-mixed with seminal plasma), normal semen (eusperm and parasperm re-mixed with seminal plasma), and natural semen (unmodified semen). Parasperm semen traveled more than 1.5 times as far as the eusperm semen. The lateral dispersion width of normal semen after emission was significantly narrower than that of eusperm semen. These findings indicate that parasperm can reduce the lateral dispersion and prolong the distance semen travels. The eusperm ratio in the lower portion of the emitted semen did not differ from that in the upper portion, indicating that eusperm evenly distribute within the ejaculate and reach the lower portion of semen. Since males cannot closely approach eggs that are deposited in the narrow space between the female's belly and the spawning substrate, restraint of lateral dispersion and prolongation of the distance semen traveled would increase the number of eusperm arriving at eggs, despite reduction of eusperm in the ejaculate.