Effects of ovalbumin on the motility and fertilising ability of fowl spermatozoa stored for 24 H at 4 degrees C

1. The effects of 0.1, 0.5, 1 and 3 g ovalbumin/100 ml diluent on spermatozoa stored for 24 h at 4 degrees C in TES diluent or BPSE diluent with or without seminal plasma were studied. 2. There was no effect of ovalbumin on the motility of spermatozoa stored in whole semen, except at an incorporation rate of 3 g/100 ml, when motility was reduced for spermatozoa stored in TES diluent. When sperm were stored in the absence of seminal plasma, ovalbumin stimulated motility but this effect was transitory. 3. The effects of ovalbumin on fertilisation rates were diluent- and concentration-dependent. Ovalbumin concentrations of 0.1 and 0.5 g/100 ml increased the fertilising ability of whole semen stored in TES diluent but 1 g/100 ml ovalbumin decreased it. However, 0.5 g ovalbumin/100 ml had no effect on the fertilising ability of spermatozoa stored in BPSE diluent, irrespective of the presence or absence of seminal plasma.     

'Serum like' albumin of fowl seminal plasma and effects of albumin on fowl spermatozoa stored at 4 degrees C.

1. Using immunoelectrophoretic and immunodiffusion methods, serum-like albumin was detected in fowl seminal plasma. Immunodiffusion showed seminal plasma albumin concentration to be 4 mg/ml, corresponding to half of the total proteins (8 mg/ml). 2. Replacing seminal plasma with a diluent containing either 1, 4, or 16 mg/ml albumin increased motility of spermatozoa stored for 24 h at 4 degrees C, 16 mg/ml being the more effective dose. 1 and 4 mg/ml had no effect on the fertilising ability of fowl spermatozoa stored for 24 h at 4 degrees C in both young (28-35 weeks old) and old birds (50-55 weeks old). 16 mg/ml albumin had no effect on fertilising rates in young but depressed it in old birds. 3. These results indicate that seminal plasma albumin may be one of the mobility stimulating factors of seminal plasma. However it does not protect fertilising ability better than the diluent alone.     

日粮补锌对荷斯坦种公牛精液品质的影响

试验选用６头荷斯坦种公牛,研究了锌对性反射时间、精液品质、精清酶活性及血、精清锌与睾酮浓度的影响。结果表明：添加锌能显著提高精液的鲜精活力、精子密度、冻后精子活力和顶体完整率（Ｐ＜０．０５）;性反射时间明显减少（Ｐ＜０．０１）;鲜精精清中ＡＫＰ和ＧＯＴ活性组间差异不显著,但ＬＤＨ活性２组显著高于１组（Ｐ＜０．０５）;冻后精清ＡＫＰ、ＧＯＴ和ＬＤＨ活性组间差异不显著（Ｐ＞０．０５）。添加锌能显著提高血清锌浓度（Ｐ＜０．０１）,但对精清锌含量的影响不明显（Ｐ＞０．０５）。血清睾酮浓度１组显著高于２组（Ｐ＜０．０５）,而精清睾酮浓度组间差异不显著（Ｐ＞０．０５）。     

Effect of seminal plasma concentration and various extenders on postthaw motility and glass wool-Sephadex filtration of cryopreserved stallion semen

OBJECTIVE: To compare the effect of semen extender and seminal plasma on postthaw motility and filtration through a glass wool-Sephadex (GWS) filter for frozen stallion semen. SAMPLE POPULATION: 7 stallions from which we collected > or = 3 ejaculates/stallion. PROCEDURES: 4 experiments were conducted to evaluate postthaw quality of frozen stallion semen. Kenney extender was compared with glucose-EDTA extender by use of various dilution rates that resulted in differing concentrations of seminal plasma. Stallions known to produce semen with poor postthaw quality were used to investigate whether a particular extender or dilution rate could improve ability of such semen to survive freeze-thaw procedures. RESULTS: Use of Kenney extender as the centrifugation extender significantly improved postthaw motility and GWS filtration, compared with glucose-EDTA. Extending semen at a dilution of 1:3 was significantly better than 1:1 for both motility and GWS filtration. In addition, including seminal plasma at a concentration of 5% in the cryopreserved semen resulted in significantly higher yield of spermatozoa after GWS filtration, compared with complete removal of SP or use of seminal plasma at 25%. Lastly, semen with poor postthaw quality had significantly improved postthaw quality in regard to motility and GWS filtration when semen was frozen with seminal plasma at a concentration of 5%, compared with semen frozen with seminal plasma at a concentration of 25%. CONCLUSIONS AND CLINICAL RELEVANCE: Use of Kenney extender at a high dilution (> or = 1:3) immediately after collection of semen can improve postthaw quality of frozen stallion semen.     

Influence of chicken native breeds on some physical and biochemical characteristics and short-term storage of semen

1. The major objective of this study was to examine the influence of 24-h storage of semen at low temperature on semen characteristics and fertilising ability of spermatozoa in two native breeds (Kadaknath-KN, Aseel Peela-AP) and White Leghorn (WL) chicken. 2. Various physical and biochemical properties of freshly ejaculated semen of KN and AP were investigated. Fertility was examined in freshly-ejaculated as well as 24-h-stored (3°C) semen diluted (1:3) with Beltsville Poultry Semen Extender. 3. No significant difference was observed in sperm motility among the different breeds whereas live counts were higher in WL than the native breeds. Body weight, semen volume and sperm concentration were highest in AP, followed by KN and WL. A similar trend was observed in the percentage of dead and morphologically-abnormal spermatozoa. 4. The activity of acid and alkaline phosphatase in seminal plasma were higher in WL than KN, whereas the opposite trend was recorded for glutamic oxaloacetic and pyruvic transaminases. The cholesterol content of semen was highest in AP, followed by KN and WL. Cholesterol was much lower in seminal plasma compared with whole semen but there were no differences between breeds. Mean values of the methylene blue reduction time test were higher in WL than in the native breeds. 5. Fertility and hatchability, using freshly-diluted semen, were poorer in the native breeds than in WL. The pattern of fertility deteriorated further, especially in native fowls, when the birds were inseminated with 24-h-stored semen. 6. In conclusion, variation in physical and biochemical characteristics of semen in native breeds compared to WL correlated with poor fertility after short-term storage of semen.     

A Comparison of the Ability of Three Commercially Available Diluents to Maintain the Motility of Cold Stored Stallion Semen

The objective of this study was to compare the ability of three commercially available extenders to promote poststorage motility of stallion spermatozoa stored at 5°C with and without centrifugation to remove the seminal plasma. Diluents tested included skim milk glucose (SKMG), INRA 96, and VMD-Z. All diluents were tested with (-SP) and without (+SP) centrifugation to remove most of the seminal plasma. In experiment I, after 48 and 72 hours of storage, total (TM) and progressive (PM) motility values were higher (P ≤.05) for those aliquots subjected to the INRA 96-SP as compared with either SKMG treatment. After 72 hours of storage, PM of spermatozoa stored in VMD-Z-SP was superior to that of spermatozoa stored in SKMG regardless of the presence of seminal plasma (P ≤.05). In the second experiment, after 48 hours of storage, PM of spermatozoa subjected to the INRA 96-SP and VMD-Z-SP treatments were superior (P ≤.05) to those for all treatments that had been stored without removal of seminal plasma. Removal of the seminal plasma and resuspension of the sperm pellet with either INRA 96 or VMD-Z resulted in TM after 48 hours of storage that were similar to those obtained after 24 hours of storage.     

Effect of phenylephrine, ergonovine, oxytocin and norepinephrine as an extender ingredient on viability of bovine spermatozoa

We evaluated effects of three concentrations of phenylephrine, ergonovine, oxytocin and norepinephrine (myometrial stimulants) on viability of spermatozoa when they were included in a seminal extender. Using a split ejaculate technique, ejaculates from each of 10 bulls were extended in egg-yolk citrate to a final concentration of 35 x 10(6) sperm/ml, including 20 mg/ml, 2 mg/ml and .2 mg/ml, of phenylephrine or ergonovine, 20 IU/ml, 2 IU/ml and .2 IU/ml oxytocin or 200 micrograms/ml, 20 micrograms/ml and 2 micrograms/ml norepinephrine prior to freezing. Extended semen without a myometrial stimulant served as the control. Percentage of intact acrosomes was determined prior to freezing for all treatments. Motility and percentage of intact acrosomes were determined immediately after thawing (0 h) and again after 4 h incubation at 37 degrees C. Percentage of intact acrosomes was reduced (P less than .05) prior to freezing by phenylephrine (20 mg/ml) and ergonovine (20 mg/ml) (phenylephrine = 56%; ergonovine = 63%; control = 74%). The same doses of phenylephrine and ergonovine reduced (P less than .05) post-thaw motility and percentage of intact acrosomes at both 0 and 4 h compared with controls. Sperm exposed even to the intermediate concentration (2 mg/ml) of ergonovine had lower (P less than .05) motility 4 h post-thaw. No other compound or concentration of compound or concentration of compound affected percentage of intact acrosome or motility. There were no two or three-way bull x compound and concentration interactions (P greater than .2).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of different processing techniques on motility and acrosomal integrity of cold-stored stallion spermatozoa

Two experiments were conducted to test whether stallionand/or semen processing techniques influenced spermatozoal motility and acrosomal status following cold storage. Ejaculates from each of 18 stallions (N=54) were collected and split. In Experiment I, a skim milk-glucose extender (SKMG) was added to the semen following a 5, 15 or 30 minute delay post-collection. Following each delay, sperm were packaged at a final concentration of 25 million progressively motile sperm per ml (PMS/ml) in a commercially available skim milk-glucose extender (SKMG). In Experiment II, sperm were packaged at concentrations of 25, 50, and 75 million PMS/ml both in the presence and absence of seminal plasma (SP) utilizing SKMG and SKMG plus PBS, respectively. In both experiments, aliquots were cooled, stored, and the percentage of progressively motile and acrosome intact spermatozoa were determined at 24 and 48 hours post-collection. In Experiment 1, delayed dilution resulted in a lower recovery of PMS. In Experiment II, removal of SP resulted in higher percentages of PMS following cold storage. Increasing the concentration of spermatozoa during packaging decreased the percentage of PMS; however, removal of SP reduced the harmful effects on spermatozoa motility. These data suggest that reducing the time that spermatozoa remain in an undiluted state and removal of SP maximize recovery of progressively motile, acrosome-intact spermatozoa. In addition, individualizing the processing techniques for each stallion may enhance spermatozoal survival following cold storage.     

Influence of the Freezing Technique (Nitrogen Liquid vs Ultrafreezer of −152°C) and Male-to-Male Variation Over the Semen Quality in Canarian Mastiff Breed Dogs

This study was carried out to assess the in vitro quality of canine semen frozen in an ultrafreezer at -152 degrees C and to evaluate the male-to-male variation of frozen semen in five male dogs of the Canarian Mastiff breed. Four ejaculates of each dog were processed individually (5% glycerol and 0.5% Equex) to reach a final concentration of 100 x 10(6) spermatozoa/ml. Then, two freezing techniques were tested to assess the seminal quality (sperm motility, live spermatozoa and abnormal sperm cell percentages) at 1, 30, 60, 120 and 360 days after freezing: (i) semen was frozen and stored in liquid nitrogen; (ii) semen was frozen and stored in the ultrafreezer at -152 degrees C. After freezing-thawing, both freezing protocols showed no significant differences in sperm motility and the percentages of live and abnormal spermatozoa. On the other hand, the microscopic characteristics of spermatozoa in fresh semen were practically similar among males; however, after the semen processing and freezing, significant differences were observed (p < 0.05) among males, especially as regards sperm motility. This inter-individual variability was detected in both freezing protocols, showing that the male-to-male variation in the seminal quality post-freezing was independent of the freezing technique used. The in vitro results obtained in the Canarian Mastiff breed confirmed that the use of ultra-freezers at -152 degrees C is a potential alternative to liquid nitrogen for storing canine semen for long periods of time.     

Autologous Whole Ram Seminal Plasma and its Vesicle-free Fraction Improve Motility Characteristics and Membrane Status but not In Vivo Fertility of Frozen–Thawed Ram Spermatozoa

Motility characteristics (assessed subjectively and with computer-assisted semen analysis) and membrane status (after staining with chlortetracycline) of washed and non-washed frozen-thawed ram spermatozoa were evaluated after incubation in buffer and buffer containing autologous whole seminal plasma or one of its two fractions: the pellet of membrane vesicles obtained by ultracentrifugation (and used at three times normal protein concentration) or the vesicle-free supernatant fraction. Whole seminal plasma and supernatant, but not membrane vesicles, improved the motility characteristics of spermatozoa after 3 and 6 h of post-thaw incubation compared with the control buffer. Resuspension and incubation with whole seminal plasma, supernatant or membrane vesicles lowered the proportion of acrosome-reacted frozen-thawed spermatozoa compared with the control buffer. Unwashed frozen-thawed semen from three rams, incubated with autologous whole seminal plasma or its fractions and inseminated using cervical or intrauterine artificial insemination, had no effect on pregnancy rates of ewes in synchronized oestrus. However, fertility was higher after laparoscopic than cervical insemination (44.9 vs 12.3%, p < 0.001). In conclusion, resuspension and incubation of frozen-thawed ram spermatozoa in autologous whole seminal plasma or its vesicle-free supernatant fraction improved their motility characteristics and, with membrane vesicles, membrane status, but these benefits were not reflected in improved fertility after cervical or intrauterine insemination.     

Effect of heterologous seminal plasma and semen extenders on motility of frozen-thawed ram spermatozoa

Ram seminal plasma increases the fertility of frozen-thawed ram spermatozoa deposited into the cervix. The aim of the current study was to compare the effect of ram seminal plasma to that of bull seminal plasma, dog prostatic fluid, protein-free TALP TrilEq (Triladyl with 0.5 mt of Equex STM paste added to each 100 mt) and heat-treated skim milk on longevity and percentages of progressively motile and aberrantly motile frozen-thawed ram spermatozoa. Three ejaculates from each of 6 rams were extended in TrilEq, pooled and frozen in straws as a single batch per ram. One hundred and eight straws (3 straws from each ram for each fluid) were thawed in random order. Once thawed, a straw was emptied into a tube with 0.85 ml of the appropriate fluid at 37 degrees C and kept at that temperature for 6 h. Motility was assessed at x200 magnification immediately (time zero) and 2, 4 and 6 h after thawing. Progressive motility decreased from each time to the next (P < 0.05) and was 39.0 % (0 h), 26.0 % (2 h), 19.6 % (4 h) and 12.6 % (6 h); SEM 1.24, n = 108 for each group. Ram seminal plasma resulted in higher progressive motility than bull seminal plasma, lower than milk, and similar to the other fluids. Ram seminal plasma resulted in lower aberrant motility than protein-free TALP and similar aberrant motility to other fluids. The effect of ram seminal plasma and dog prostatic fluid was very similar. The effect of ram seminal plasma on the fertility of frozen-thawed ram spermatozoa deposited into the cervix is not due an exceptionally beneficial effect on the motility of spermatozoa.     

Biochemical observations on beagle dog semen.

Semen samples were collected at weekly intervals for six weeks from eight sexually mature beagles previously shown to produce normal ejaculates. Seminal plasma and sperm fractions were separated by centrifugation and the sodium, potassium, alanine and aspartate aminotransferases, acid and alkaline phosphatase concentrations in the two fractions determined. Regression analysis of the mean weekly values obtained from physical and biochemical examination of the ejaculates showed that sodium ion concentration was highest in seminal plasma. The highest levels of aminotransferases were found in sperm fractions. Those enzymes may be indices of abnormal or damaged spermatozoa. Acid and alkaline phosphatase activity was 100 times greater in seminal plasma than in sperm fractions. Phosphatase concentrations are likely to be dependent on prostate activity. Measurement of acid phosphatase in canine semen therefore may be a useful index of prostate function. The motility of the semen samples was independent of the potassium concentration in seminal plasma. However, there was some evidence of a correlation between sperm motility and the enzyme and sodium content of seminal plasma.     

Exploring the metabolome of seminal plasma in two different horse types: Light versus draft stallions

The application of the ‘omics’ studies in the field of animal reproduction has been aimed at identifying novel biomarkers of fertility since the last few years. When assessing reproductive efficiency in horses, breed should also be taken into account as it can influence semen quality and fertility. Considering the growing interest in metabolomic analysis to evaluate male fertility, we aimed to investigate the metabolomic profile of seminal plasma in two different horse breeds. Twelve healthy stallions, n.6 American Quarter Horse (AQH) and n.6 Italian Draft Horse (IDH) stallions, regularly used for artificial insemination, were included in the study. Two semen collections, performed 30-day apart, were considered for the assessment of semen parameters including gel-free volume, spermatozoa (spz) concentration, spz progressive motility and seminal plasma analysis by ¹H-NMR.Semen characteristics differed between IDH and AQH (p < .05) as well as the first cycle conception rate that was higher in AQH than IDH (p = .001). Metabolomic analysis quantified 56 molecules in equine seminal plasma, with 11 metabolites showing different concentrations in IDH compared to AQH (p < .05).This study provided evidence of differences in seminal plasma metabolites' concentrations between studied horse types, highlighting specific metabolomic fingerprints characterizing AQH and IDH sperm.     

Effect of semen collection in extender solution on the characteristics of goat spermatozoa

The objective of this experiment was to investigate whether the motility parameters and acrosome integrity of goat ejaculated spermatozoa are affected by collecting semen into tubes containing an extender, and thereby determine the significance of reducing contact between seminal plasma and the sperm membrane at ejaculation. Semen were collected from three goats into tubes containing 0, 1 or 10 ml extender, or collected into tubes containing 10 ml extender supplemented with 0.1, 1 or 5% BSA. Sperm motion parameters were evaluated immediately after collection, after washing, and during a 3-h thermal resistance test. Acrosome integrity was assessed using FITC-PNA staining. Semen collection into tubes containing 10 ml extender produced higher sperm motility, progressive motility, and acrosome integrity than that using a smaller volume of extender. Furthermore, collection into 5% BSA-containing extender exhibited higher sperm characteristics and maintained high sperm motility and progressive motility throughout incubation. In conclusion, semen collection into tubes with a large volume of extender, especially extender containing higher concentrations of BSA, improved the quality of ejaculated spermatozoa, strongly suggesting that the in vitro functional characteristics of the spermatozoa were abruptly modified by flash sperm contact with accessory sex gland fluid at ejaculation.     

Prostasomes of Canine Seminal Plasma – Zinc‐Binding Ability and Effects on Motility Characteristics and Plasma Membrane Integrity of Spermatozoa

Prostasomes are small lipid membrane‐confined vesicles that are involved in various fertilization‐related processes. The aim of this study was to demonstrate canine seminal plasma prostasomes' ability to bind zinc ions, as well as examining their effects on sperm motility characteristics and plasma membrane integrity during cold storage. Ejaculates, collected from five cross‐bred dogs (n = 50), were subjected to ultracentrifugation followed by gel filtration (GF) on a Superose 6 column. Prostasomes appeared as a single fraction in the elution profile. Transmission electron microscopy (TEM) analysis of canine prostasomes revealed the presence of membrane vesicles with diameters ranging from 20.3 to 301 nm. The zinc‐affinity chromatography on a Chelating Sepharose Fast Flow – Zn^{2 +} showed that from 93 to 100% of the prostasome proteins bind zinc ions (P⁺Zn). SDS‐PAGE revealed that canine P⁺Zn comprised four protein bands, with low molecular weights (10.2–12 kDa). We have also shown a positive effect of prostasomes (p < 0.05), especially variant B (2% of total seminal plasma protein) on canine sperm motility parameters after 2 h storage at 5°C (TMOT%, 44.75 ± 5.18) and PMOT%, 12.42 ± 1.59) and VAP, VSL, VCL, when compared with Control (TMOT%, 7.30 ± 1.41 and PMOT%, 1.70 ± 0.42). Higher percentage of spermatozoa with intact plasma membrane (SYBR/PI dual staining) and intact acrosome (Giemsa stained), after 2 h storage at 5°C, was showed, in variant A (1.5% of total seminal plasma protein) and B, when compared with Control and variant C (2.5% of total seminal plasma protein). The prostasomes' effect on motility and plasma membrane integrity of canine cold‐stored spermatozoa may be related to their ability to bind zinc ions and regulate their availability to the sperm.     

Evaluation of spermiation indices with multiple sperm collections in endangered sterlet (Acipenser ruthenus)

This study investigated the effects of multiple collections of sperm on endangered sterlet (Acipenser ruthenus) sperm functional parameters [spermatozoa motility and curvilinear velocity (VCL)] as well as on protein concentration and osmolality of seminal plasma. The average sperm volume and mean spermatozoa concentration per male were significantly altered with multiple collections. On the other hand, no significant effect of multiple collections on protein concentration of seminal plasma was observed. In all experimental groups, moderate impact of sequential collection on osmolality (p < 0.05) of seminal plasma was observed. Ninety to 100% of motile spermatozoa were observed at 15 s after activation, with an average VCL of 181.12 ± 19.10 μm/s. After 90 s, average VCL decreased to 130 ± 26 μm/s. Motility was maintained for up to 4 min. The maximum percentage of motile spermatozoa was observed after the third collection of sperm. The spermatozoa VCL increased significantly with subsequent collections. The results of this study provide new data on the effects of multiple collections on quantitative and qualitative parameters of sperm in sterlet. The data confirmed that the sequential stripping has no negative effect on the percentage of motility and spermatozoa velocity. This should be beneficial for the development of sterlet aquaculture programs.     

The effect of vitamin a supplementation on seminal characteristics and vitamin a absorption in stallions

Fourteen mature stallions were paired based on age and pretreatment spermatozoal output. One member of each pair was assigned to either 1) control (3 ml corn oil) or 2) treated (132,000 IU retinyl palmitate in 3 ml corn oil) experimental groups. Treatments were added to oat rations every other day. Seminal characteristics (gel free volume, gel volume, total seminal volume, percent progressively motile spermatozoa, number of spermatozoa per ml, percentage morphologically normal spermatozoa and spermatozoal membrane stability) and total scrotal width of each stallion were recorded before (February) and after three months of vitamin A supplementation (June). Plasma vitamin A was measured at 0,6,12,24, and 48 hours following the first and last treatments to document absorption. There were no treatment effects (p>.05) on seminal characteristics or scrotal width. Seasonal increases were recorded in gel-free volume, total seminal volume, percent spermatozoal motility, total spermatozoal output, percentage of morphologically normal spermatozoa, and total scrotal width. Plasma vitamin A was lower during the second collection period (June) than the first (February) in both treatment groups. Peak plasma vitamin A was observed 48 hours following ingestion of the first dose of the vitamin but at 12 hours following the last treatment.     

Assessment of copper and zinc status of farm horses and training thoroughbreds in south-east Queensland

The copper and zinc concentrations in the blood of stabled thoroughbred horses and in Australian Stock Horses mares at pasture, either late pregnant or lactating were determined by an atomic absorption spectroscopic method. The plasma concentration of the trace elements in these apparently normal horses were generally below the "normal" range. The plasma copper, caeruloplasmin copper, whole blood copper and plasma zinc concentrations in the stabled thoroughbreds were 0.76 +/- 0.19 micrograms/ml (n = 82), 0.56 +/- 0.14 micrograms/ml (n = 83), 0.75 +/- 0.18 micrograms/ml (n = 82) and 0.47 +/- 0.09 micrograms/ml (n = 83) respectively. The plasma copper and zinc concentrations of all the brood mares at pasture (pregnant and lactating) were 0.56 +/- 0.20 micrograms/ml and 0.47 +/- 0.11 micrograms/ml (n = 30). The plasma copper concentration of the pregnant group of mares (0.64 +/- 0.18 micrograms/ml; (n = 14) was greater than that of the lactating mares (0.49 +/- 0.21; (n = 16). Variation in the plasma copper concentration was also identified between stabled and farm horses, between horses of different stables and between horses of different ages. The proportion of plasma copper bound to caeruloplasmin was 73 +/- 11.8%. These low concentrations of copper and zinc in the plasma of apparently normal horses are of clinical significance since recent evidence has indicated that copper deficiency appears to promote the development of skeletal abnormalities in foals. An alternative to the use of a single plasma sample to identify the copper or zinc deficient horse was discussed.     

Effect of seminal plasma fractions on stallion sperm survival after cooled storage

This study aimed to evaluate stallion sperm survival after 24 h of cooled storage in the presence of seminal plasma (SP) derived from the sperm-rich fractions (SRF) or sperm-poor fractions(SPF) of the ejaculate, without SP, or in the presence of SP from other stallions. Ejaculates were collected from four stallions using an automated phantom, which separated the semen into five cups. Centrifuged and washed spermatozoa from cup 2 (SRF) were mixed with skim milk extender to a concentration of 100 x 10(6) sperm/ml and then 1:1 (v/v) with SP from the stallion's own or another stallions' second (SP-SRF) or last cup (SP-SPF). Skim milk extender (K) and skim milk extender supplemented with modified Tyrode's medium (KMT) were used as control treatments. After a 24-h storage period in a transport container, spermatozoa were evaluated for motion characteristics and plasma membrane integrity by calcein acetoxymethyl (AM)/propidium iodide staining. The percentage of spermatozoa with intact plasma membranes after storage was lower in SP-SRF than in SP-SPF, and the highest in K (P < 0.05). Progressive motility (PMOT) was lower for sperm stored in SP-SRF than for sperm stored in SP-SPF (P < 0.05), but there was no significant difference in total motility (TMOT). Sperm stored in KMT (P < 0.05) registered the highest TMOT and PMOT percentages. Osmolarity was significantly higher and pH lower in K than in KMT or SP. Treatment with SP-SPF from three stallions benefited the PMOT of sperm from one stallion. These preliminary findings suggest that SP from SRFs may be more harmful during storage than SP from SPFs. Removal of SP improves sperm survival in KMT extender, and exchanging SP between stallions seems to influence sperm survival.     

Distinct Effects of Boar Seminal Plasma Fractions Exhibiting Different Protein Profiles on the Functionality of Highly Diluted Boar Spermatozoa

The aim of this study was to evaluate how different protein profiles of seminal plasma (SP) fractions affect sperm functionality in vitro. Ejaculates from three boars were separated into six fractions. The fractions differed from each other in their sperm content, in their total SP protein content, and their spermadhesin PSP-I/PSP-II and heparin-binding protein (HBP) concentrations. Spermatozoa were mainly recovered in fraction 2 (sperm-rich fraction, >1800 × 10⁶ spermatozoa/ml), whereas the pre-sperm fraction 1 and the post-sperm fractions 4–6 contained low numbers of spermatozoa (<500 × 10⁶/ml). Except in fraction 2, the total SP protein concentration and the concentration of both, spermadhesin PSP-I/PSP-II and the HBPs increased with fraction order. Distinct time-dependent effects were observed on motility characteristics and membrane integrity of highly diluted boar spermatozoa upon incubation with a 10% dilution of the SP from each fraction. The highest sperm viability was recorded after exposure for 5 h to fraction 2, followed by fractions 1 and 3. The percentages of motile spermatozoa also differed significantly among fractions after 5 h of incubation. Spermatozoa incubated with SP of fractions 1–3 showed the highest percentage motility. We conclude that different SP fractions exert distinct effects on the functionality of highly diluted boar spermatozoa. Fractions 1–3 appear to promote sperm survival, whereas fractions 4–6 seem to be harmful for preserving the physiological functions of highly diluted boar spermatozoa.