Effect of dietary alpha-tocopheryl acetate and ascorbic acid on rabbit semen stored at 5 degrees C

The objective of this research was to verify the effects of dietary alpha-tocopheryl acetate (50 vs. 200 mg/kg diet) and ascorbic acid (0 vs. 1 g/L water) on the relative amounts on semen and motion characteristics, oxidative stability and fertilizing ability of rabbit spermatozoa stored for 24 h at 5 degrees C. A high amount of dietary (alpha-tocopheryl acetate significantly increased the level of Vitamin E in the semen (0.90 vs. 0.41 micromol/L) and its oxidative stability after storage (Thiobarbituric Acid Reactive Substances-- TBARS 15.88 vs. 20.90 nmol Malondialdehyde--MDA/mL). Ascorbic acid showed a different effect in relation to the Vitamin E status of animals: when associated with the higher level of Vitamin E it increased the (alpha-tocopherol and the oxidative stability of semen (2.67 micromol/L and 12.25 nmol MDA/mL, respectively), whereas both parameters were reduced with lower Vitamin E (0.13 micromol/L and 21.20 nmol MDA/mL). Semen traits were not modified by the separate supplementation of supranutritional levels of vitamins, whereas their combination significantly improved the viability and the kinetics of spermatozoa (e.g. track speed: 95.13 vs. 71.31 microm sec(-1)) with an increase in fertility rate (70.0 vs. 63.3; P=0.06) that could be considered almost significant.     

Oxidative status and semen characteristics of rabbit buck as affected by dietary vitamin E,C and n-3 fatty acids

The aim of this study was to investigate the effect of dietary supplementation of long chain fatty acids (C > or = 20 LCP) of the n-3 series, vitamin E and vitamin C on the antioxidant capacity of rabbit buck and on semen characteristics. Fifty male rabbits at 30 days were randomly assigned to five different diets: Control (50 mg x kg(-1) diet alpha-tocopheryl acetate), Vitamin E (200 mg x kg(-1) diet alpha-tocopheryl acetate), n-3 (2% ROPUFA oil + 50 mg x kg(-1) diet alpha-tocopheryl acetate), n-3 + E (2% ROPUFA oil + 200 mg x kg(-1) diet alpha-tocopheryl acetate) and n-3 + E C (2% ROPUFA oil + 200 mg x kg(-1) diet alpha-tocopheryl acetate + 0.5 g x L(-1) vitamin C in the drinking water). The levels of vitamins E and C and reactive oxygen metabolites (ROMs) in the blood plasma were evaluated at different ages. The antioxidant capacity and ROMs of seminal plasma, the fatty acid profile of sperm phospholipids, the semen traits and the oxidative processes during storage (24 h at + 4 degrees C) were carried out weekly for 5 wk starting from the 5th month of age. Vitamin E addition showed enough antioxidant protection only when associated with no lipid enriched diets. The n-3 supplementation modified the fatty acid profile of the spermatozoa membrane and simultaneously enhanced oxidative processes. Only the association with supranutritional levels of vitamins E and C inhibited the oxidative processes and improved the characteristics of fresh and stored rabbit semen.     

Impact of changes in composition of storage medium on lipid content and quality of turkey spermatozoa

Turkey semen quality is damaged by long term in vitro storage. The objective of the present study was to determine whether changes in energy substrates and antioxidants of semen extender could limit loss of quality and lipid content of turkey spermatozoa during storage. Spermatozoa were incubated in extenders based on Beltsville Poultry Semen Extender (BPSE) to which different energy substrates (acetate, pyruvate and hydroxybutyric acid) or antioxidant (Vitamin E) had been added. Semen was stored at 4 degrees C for 48 h and changes in quality, phospholipid and malondialdehyde (MDA) content of semen were evaluated. Among the different substrates studied, only acetate was able to limit the loss of motility and ATP content after 48 h in vitro storage. Losses of spermatozoal phospholipids were similar when gametes were incubated in an extender without any substrate or in normal BPSE (784-675nmol/10(9) spz versus 837-703 nmol/10(9) spz). However, motility and ATP content were significantly more affected after 48 h of storage in samples incubated without substrates than in BPSE (motility, 2.2 versus 0; ATP, 10 nmol/10(9) spz versus 3 nmol/10(9) spz). The addition of Vitamin E to the extender did not modify either the MDA or phospholipid content of fresh or stored spermatozoa, but increased the motility of stored semen. In conclusion, acetate is an essential substrate for in vitro storage. Spermatozoal phospholipids decreased during storage, but this did not seem to originate from metabolism of endogenous fatty acids. The positive effects of Vitamin E on semen storage did not originate from preservation of lipid oxidation.     

Effect of organic selenium on turkey semen quality during liquid storage

The aim of the present study was to evaluate the effect of dietary organic selenium on the turkey semen during storage. Twenty males (BUT, Big 6, 40 weeks of age) were divided into control (n = 10) and experimental group (n = 10). The turkeys in the both groups were fed with a commercial diet containing 0.1 ppm Se in the form of sodium selenite. The experimental birds were additionally supplied with 0.3 ppm organic Se in the form Sel-Plex™ (Alltech, Inc.). After 30 days of feeding, the semen samples were collected twice a week for the 3 weeks of the study and diluted 1 + 1 (v/v) with TUR-2 diluent, and stored in a water bath (+10 to 15° C) for 6 h. The percentage of motile spermatozoa, the sperm viability (live/dead spermatozoa), total lipids, phospholipids and total cholesterol were assessed in fresh and stored semen. The fertilizing ability of semen was assessed by artificial insemination of 30 hens per group with dose containing 200 × 10⁶ spermatozoa weekly. After 6 h of semen storage, the motility of spermatozoa decreased significantly in the control group (by 8.7 relative percent, P < 0.05) and only by four relative percent (P > 0.05) in experimental group reflecting a protective effect of dietary Se supplementation. The proportion of live spermatozoa was higher in fresh semen and significantly lower in stored semen. The positive effect of Se supplementation was observed on the lipid composition of stored semen: the concentration of the total lipids and phospholipids in the seminal plasma from control group significantly increased, while in the experimental group remained constant. Better semen integrity in the experimental group was associated with an improved fertilizing ability of spermatozoa: the fertility rate of stored spermatozoa in the control group was 88%, while in the experimental group was 90.5%.     

Oxidative and nitrosative stress markers in bus drivers

Exposure to ambient air pollution is associated with many diseases. Oxidative and nitrosative stress are believed to be two of the major sources of particulate matter (PM)-mediated adverse health effects. PM in ambient air arises from industry, local heating, and vehicle emissions and poses a serious problem mainly in large cities. In the present study we analyzed the level of oxidative and nitrosative stress among 50 bus drivers from Prague, Czech Republic, and 50 matching controls. We assessed simultaneously the levels of 15-F(2t)-isoprostane (15-F(2t)-IsoP) and 8-oxodeoxyguanosine (8-oxodG) in urine and protein carbonyl groups and 3-nitrotyrosine (NT) in blood plasma. For the analysis of all four markers we used ELISA techniques. We observed significantly increased levels of oxidative and nitrosative stress markers in bus drivers. The median levels (min, max) of individual markers in bus drivers versus controls were as follows: 8-oxodG: 7.79 (2.64-12.34)nmol/mmol versus 6.12 (0.70-11.38)nmol/mmol creatinine (p<0.01); 15-F(2t)-IsoP: 0.81 (0.38-1.55)nmol/mmol versus 0.68 (0.39-1.79)nmol/mmol creatinine (p<0.01); carbonyl levels: 14.1 (11.8-19.0)nmol/ml versus 12.9 (9.8-16.6)nmol/ml plasma (p<0.001); NT: 694 (471-3228)nmol/l versus 537 (268-13833)nmol/l plasma (p<0.001). 15-F(2t)-IsoP levels correlated with vitamin E (R=0.23, p<0.05), vitamin C (R=-0.33, p<0.01) and cotinine (R=0.47, p<0.001) levels. Vitamin E levels also positively correlated with 8-oxodG (R=0.27, p=0.01) and protein carbonyl levels (R=0.32, p<0.001). Both oxidative and nitrosative stress markers positively correlated with PM2.5 and PM10 exposure. In conclusion, our study indicates that exposure to PM2.5 and PM10 results in increased oxidative and nitrosative stress.     

Effect of dietary antioxidant supplementation on fresh semen quality in stallion

In this study, the effect of dietary supplementation of organic selenium, vitamin E, and zinc on raw semen characteristics was evaluated. Ten stallions with normal fertility were divided into two groups: a control group (CG), in which standard diet was provided, and a treated group (TG), in which the standard diet was supplemented with 1500 mg of α-tocopherol acetate, 360 mg of zinc, and 2.5 mg of organic selenium on a daily basis. Semen parameters on fresh semen were evaluated three times in all stallions before antioxidant supplementation (T0) and 30 (T1), 60 (T2), and 90 (T3) d after supplementation. Dietary supplementation with experimental antioxidants resulted in a significant increase in average path velocity (121.9 ± 3.1 μm/sec in TG vs 118.9 ± 4.3 μm/sec in CG), straightness (86.2 ± 2.4 % vs 82.6 ± 3.9 % in TG and CG respectively), viability (75.6 ± 10.2 % in TG vs 72.3 ± 6.9 % in CG) and total seminal plasma antioxidants levels (2.7 ± 0.5 mmol/l vs 1.9 ± 0.4 mmol/l in TG and CG respectively) while progressive motility 69.7 ± 11 % vs 62.2 ± 9.3 % in TG and CG stallions respectively) and abnormal sperm morphology (8.2±1.5 % in TG vs 14.4±4 % in CG) significantly improved in treated stallions after 60 d of supplementation. In contrast with previously reported in other species, a negative effect of antioxidant supplementation on semen concentration was recorded in the TG. A positive correlation between progressive motility and total antioxidants in seminal plasma in both treated and control stallions suggested that motility is affected by oxidative-antioxidative status, and that dietary antioxidant supplementation could increase the ability of spermatozoa to contrast reactive oxygen species or the ability of seminal plasma to reduce the oxidative stress. The improvement of semen parameters after antioxidant supplementation was not linear, and after 30 d (or 60 d for some parameters), a further increase was not noted. This evidence suggested that in our standard conditions, dietary intake of these antioxidants could be slightly under the dietary requirement and further evaluation of the actual nutrition requirements of organic selenium, zinc, and vitamin E in the stallion are needed.     

Influence of dietary alpha-tocopheryl acetate supplementation on cholesterol oxidation in retail packed chicken meat during refrigerated storage

This study examined the effect of dietary alpha-tocopheryl acetate on lipid and cholesterol oxidation in retail packed chicken meat during refrigerated storage. Male broiler chicks were randomly assigned to five pens containing 30 chicks each, which was subjected to one of five dietary treatments (0, 50, 100, 200, or 400 IU of alpha-tocopheryl acetate/kg diet). Five different levels of alpha-tocopherol were supplied to the chicks from 3 to 6 weeks. After 42 d of feeding all the broilers were slaughtered, and the carcasses were packed in polyethylene bags individually, bags similar to those used in the retail trade, and stored for 12 d at 4 degrees C. Growth performance and fatty acid composition were not affected by the dietary alpha-tocopherol levels. The alpha-tocopherol content in breast and thigh muscles increased as the level of dietary alpha-tocopherol increased. The supplementation with 200 or 400 IU of alpha-tocopherol was more effective in reducing the level of lipid oxidation (P<0.05) and total cholesterol oxidation products (P<0.05). Therefore, an increase in the dietary alpha-tocopherol level from 200 to 400 IU/kg feed causes major improvements in the oxidative stability of chicken meat during refrigerated storage.     

Selenium, glutathione peroxidases, and some other antioxidant parameters in blood of patients with chronic renal failure

In the present study several parameters associated with oxidative stress were examined in the blood of 25 chronic renal failure (CRF) patients and the results were compared with 18 healthy subjects. Mean creatinine concentration in patients was 1,216 +/- 292 micromol/l. Selenium (Se) concentration in red cells, whole blood and in plasma of CRF patients (106 +/- 32.5, 59.0 +/- 16.7 and 42.4 +/- 13.8 ng/ml, respectively) was significantly (0.0001 < P 0.01) lower (by 20-42%) compared with the controls. Red cell and plasma glutathione peroxidase (GSH-Px) activities (16.6 +/- 3.4 U/g Hb and 93.7 +/- 32.9 U/l plasma) were lower by 12 and 53% (P < 0.05 and < 0.0001, respectively) in patients than in healthy subjects. GSH concentration in red cells of patients (2.81 +/- 0.45 mmol/l) was significantly (P < 0.001) higher (by 20%) than in control group. Malonyldialdehyde (MDA) concentration (expressed as thiobarbituric acid-reactive substances) in red cells of patients (725 +/- 155 nmol/g Hb) was significantly (P < 0.001) higher (by 28%) than in control group. No significant difference was observed in the activity of superoxide dismutase in pLasma between the two groups. In conclusion, our results confirm that the aLterations in Se levels in blood components and in GSH-Px activity in plasma show that the kidney plays an important role in Se homeostasis and in plasma GSH-Px synthesis.     

Combined effect of DHA and alpha-tocopherol enrichment on sperm quality and fertility in the turkey

The aim of the present study was to characterize the dietary effects of n-3 LC-PUFA and alpha-tocopheryl acetate (vE) on the quality, phospholipid fatty acid composition, alpha-tocopherol content (alpha-T) and in vitro susceptibility to lipid peroxidation in turkey semen. Fertility of fresh semen was also evaluated. Male turkeys were randomly divided and fed either a control diet or a fish oil and vE rich diet (FO diet) from 40 to 60 weeks of age. The FO diet increased the proportion of n-3 fatty acids in spermatozoa and as a consequence the (n-3)/(n-6) ratio also increased. These changes did not affect the proportion of n-9 PUFAs, particularly of C22:3n-9, in semen. The sperm content of alpha-T was dependent by the dietary supplementation of the vitamin and the sperm content was more than doubled supplying 120 mg kg(-1) of feed to the males compared to the 60 mg kg(-1) of feed in the control diet. In agreement with the major content of alpha-T in spermatozoa collected from the FO group were significantly less susceptible to in vitro induced oxidation. The reproductive capacity of the male breeders was not affected by the diet; however the result is considered of some relevance for field conditions where even very small changes have economic interest being applied to large bird population.     

Effects of dexamethasone treatment (to mimic stress) and Vitamin E oral supplementation on the spermiogram and on seminal plasma spontaneous lipid peroxidation and antioxidant enzyme activities in dogs

The objective was to determine if treatment with dexamethasone (to mimic stress) has a deleterious effect on the spermiogram and on the composition of seminal plasma in the dog and whether adverse effects were reduced by oral supplementation with Vitamin E. Eighteen adult male Rottweiler dogs were randomly allocated in a 2 x 2 factorial treatment design (with or without dexamethasone treatment versus with or without Vitamin E supplementation). Dogs in the supplemented group received 500 mg of alpha-tocopherol (Vitamin E)/dog/day per os for 10 weeks. Dexamethasone (0.01 mg/kg/day i.m.) was given once daily for 7 days, starting 7 days after the onset of Vitamin E supplementation. Food intake, body condition score and body weight were assessed daily. Semen collections (digital manipulation) were performed twice weekly for 14 weeks and blood samples (for plasma concentrations of cortisol and testosterone) were collected once a week. Dexamethasone treatment significantly reduced ejaculate volume and increased thiobarbituric acid-reactive substances (TBARS) in the seminal plasma. In contrast, supplementation with Vitamin E increased sperm motility, vigor and concentration and decreased the percentage of major sperm defects. In conclusion, dexamethasone treatment (to mimic stress) had a deleterious effect on the spermiogram and on the seminal plasma lipid peroxidation in dogs; however, some of these effects were prevented by oral supplementation with Vitamin E.     

Freeze-thawing as the factor of spontaneous activation of spermatozoa motility in common carp (Cyprinus carpio L.)

In the present study, we investigated the possibility of spontaneous carp spermatozoa activation by freeze-thawing. To evaluate this, the parameters of spermatozoa motility percentage, velocity, ATP content level and fertility rate of sperm were used. The motility and velocity of spermatozoa activated by freeze-thawing were characterized by motile spermatozoa with a median value of 16% and a velocity of 98 μm/s. In addition, the motility and velocity of sperm from the thawed samples were significantly lower than in the control (median value of 100% for sperm motility and 175 μm/s for sperm velocity). Furthermore, a spontaneously activated spermatozoa motility terminated within five minutes post-thaw time. After freeze-thawing the ATP level significantly decreased with post-thaw time (46 nmol ATP/10⁹ and 10 nmol ATP/10⁹ at 25 s and 10 min after thawing, respectively). Fertility of spermatozoa was not significantly affected within 10 min post-thaw. On the other hand, the fertility of frozen-thawed sperm was significantly lower if compared to fresh sperm. We conclude that the freeze-thawing procedure spontaneously activated spermatozoa motility in common carp. However, this activation did not negatively affect the fertility of frozen-thawed sperm.     

Dietary vitamin E supplementation affects tissue lipid peroxidation of hybrid tilapia,Oreochromis niloticus x O. aureus

A feeding trial was conducted to evaluate the effects of dietary vitamin E contents on the growth, ascorbate induced iron-catalyzed lipid peroxidation in post-mortem muscle and liver tissue, and Raman spectral changes in lens of juvenile hybrid tilapia (Oreochromis niloticus x O. aureus). Experimental fish were fed practical diets supplemented with 0, 50, 100, 200, 450 and 700 mg alpha-tocopheryl acetate/kg diet for 14 weeks. There was no significant difference in weight gain, feed conversion ratio and protein efficiency ratio among fish fed test diets (P>0.05). Protein content of fish fed diet containing the lowest vitamin E level was the lowest (P<0.05) among all groups. No difference was found in other body constituents among test fish (P>0.05). The thiobarbituric acid-reactive substances produced by iron-catalyzed lipid peroxidation in muscle and liver tissue of fish fed the diet without alpha-tocopheryl acetate supplementation were significantly (P<0.05) greater than those from fish fed diets containing higher levels of alpha-tocopheryl acetate. Dietary vitamin E supplementation increased the antioxidant capability of tilapia tissues against lipid peroxidation. Further, dietary vitamin E supplementation also influenced the lens cortical membrane structure of tilapia.     

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.     

Increased lipid peroxidation in pregnant women after iron and vitamin C supplementation

Iron overload could promote the generation of free radicals and result in deleterious cellular damages. A physiological increase of oxidative stress has been observed in pregnancy. A routine iron supplement, especially a combined iron and vitamin C supplementation, without biological justifications (low hemoglobin [Hb] and iron stores) could therefore aggravate this oxidative risk. We investigated the effect of a daily combined iron supplementation (100 mg/d as fumarate) and vitamin C (500 mg/d as ascorbate) for the third trimester of pregnancy on lipid peroxidation (plasma TBARS), antioxidant micronutriments (Zn, Se, retinol, vitaminE, (β-carotene) and antioxidant metalloenzymes (RBC Cu-Zn SOD and Se-GPX). The iron-supplemented group (n=27) was compared to a control group (n=27), age and number of pregnancies matched. At delivery, all the women exhibited normal Hb and ferritin values. In the supplemented group, plasma iron level was higher than in the control group (26.90±5.52 mmol/L) and TBARs plasma levels were significantly enhanced (p<0.05) (3.62±0.36 vs 3.01±0.37 mmol/L). No significant changes were observed in plasma trace elements and red blood cell antioxidant metalloenzymes. Furthermore, the α-tocopherol plasma level was lowered in the iron-supplemented groups, suggesting an increased utilization of vitamin E. These data show that pharmalogical doses of iron, associated with high vitamin C intakes, can result in uncontrolled lipid peroxidation. This is predictive of adverse effects for the mother and the fetus. This study illustrates the potential harmful effects of iron supplementation when prescribed only on the assumption of anemia and not on the bases of biological criteria.     

Role of seminal plasma in damage to turkey spermatozoa during in vitro storage

In vitro storage of turkey spermatozoa is performed without consideration of the potential role of seminal plasma on sperm functions. We report the effects of seminal plasma on membrane permeability, lipid metabolism, energy status, motility and fertility of turkey spermatozoa stored at 4 or 20 degrees C. Phospholipid content (1077 nmol/10(9) spz versus 1219 nmol/10(9) spz at 48 h) and membrane permeability of spermatozoa were significantly damaged by the presence of seminal plasma after 48 h of storage at 4 degrees C, whereas damage to ATP content and fertility occurred earlier damaged by this presence (fertility after 24h storage 51% with seminal plasma versus 71% without). At 20 degrees C, seminal plasma decreased the phospholipid content of spermatozoa in the first hour of storage (1326 nmol/10(9) spz versus 1636 nmol/10(9) spz). Twenty-four hours later, this effect was masked by intense lipid peroxidation. These results show that seminal plasma is deleterious to storage of turkey spermatozoa at 4 degrees C and is involved in phospholipid metabolism of spermatozoa. Lipid peroxidation could be responsible for the acceleration of the degradation of sperm phospholipids during storage at 20 degrees C. However, lipid peroxidation seems not to be active at 4 degrees C. In this case, we suggest that phospholipase activation may contribute to sperm degradation, especially in the presence of seminal plasma.     

Performance of rabbits and oxidative stability of muscle tissues as affected by dietary supplementation with oregano essential oil

The effect of dietary supplementation with oregano essential oil on the performance of rabbits, and the susceptibility of the produced raw and thermally treated muscle tissue to lipid oxidation during refrigerated storage, were investigated. A total of 96 weaned rabbits were separated into four equal groups with three subgroups each. One group was given the basal diet and served as control, two groups were administered diets supplemented with oregano essential oil at levels of 100 and 200 mg/kg diet, whereas the remaining group was given a diet supplemented with alpha-tocopheryl acetate at 200 mg/kg. During the 42-day experimental period, body weight and feed intake were recorded weekly and the feed conversion ratio was calculated. Feeding the experimental diets to rabbits, performance parameters were not affected. Therefore, dietary oregano essential oil exerted no growth-promoting effect on rabbits. With increased supplementation of oregano essential oil, malondialdehyde values decreased in both raw and thermally treated muscles during refrigerated storage. This finding suggests that dietary oregano essential oil exerted a significant antioxidant effect. Dietary supplementation of oregano essential oil at the level of 200 mg/kg was more effective in delaying lipid oxidation compared with the level of 100 mg/kg, but inferior to dietary supplementation of 200 mg alpha-tocopheryl acetate per kg. This study indirectly provides evidence that antioxidant compounds occurring in oregano essential oil were absorbed by the rabbit and increased the antioxidative capacity of tissues.     

Antioxidant effect of vitamin E and glutathione on lipid peroxidation in boar semen plasma

The protective effect of vitamin E and reduced glutathione (GSH) against lipid peroxidation in boar semen plasma was studied. The lipid peroxidation, measured by the test for thiobarbituric acid reactive substances (TBARS), doubled in the presence of the lipid peroxidation Fe²⁺-sodium ascorbate-inducing system. The ascorbate-induced TBARS were inhibited by about 62% through the water-soluble vitamin E analog (TROLOX) and about 57% by GSH. In the in vivo experiments, 7 wk of oraldl-α-tocopherol acetate (1000 IU/d/animal) administration caused a significant fall in the level of the semen plasma TBARS, from 2.2±0.09 to 1.2±0.13 nmol MDA/mL. The semen plasma superoxide dismutase (SOD) and GSSG tended to increase with the time of vitamin E administration, but the increment did not reach a significant level by the seventh week. The vitamin E supplementation significantly increased the number of spermatozoa per 1 cm³ of ejaculate. The protective role of vitamin E and GSH with respect to boar semen against fatty acid peroxidation and a positive influence of vitamin E supplementation on semen quality have been evidenced.     

Ability to activate oocytes and chromosome integrity of mouse spermatozoa preserved in EGTA Tris-HCl buffered solution supplemented with antioxidants

Potential methods for cryopreservation of mouse spermatozoa are freeze-drying, desiccation, and suspension in EGTA Tris-HCl buffered solution (ETBS: 50 mM NaCl, 50 mM EGTA, and 10 mM Tris-HCl). To determine the duration that mouse spermatozoa suspended in ETBS-based solutions could retain their normal characteristics without freezing, spermatozoa collected from the cauda epididymis were suspended in ETBS or in ETBS supplemented with the antioxidants, dimethyl sulfoxide (DMSO), or DL-alpha-tocopherol acetate (Vitamin E acetate; VEA) diluted in DMSO, then held at ambient temperature (22-24 degrees C) for up to 9 days. When oocytes were injected with spermatozoa preserved in ETBS alone, activation rates of oocytes and chromosome integrity at the first cleavage metaphase decreased at 1 day (P < 0.001) and 2-4 days (P < 0.01) following treatment. When oocytes were injected with spermatozoa preserved in ETBS supplemented with DMSO or VEA/DMSO, chromosome integrity did not decrease significantly (through 9 days of preservation). Although DMSO maintained sperm chromosome integrity more effectively than VEA/DMSO up to 2-4 days (91 and 67%, normal karyotypes in DMSO and VEA/DMSO, respectively), VEA/DMSO helped to maintain the ability of spermatozoa to activate oocytes, but did not enhance the maintenance of sperm chromosome integrity. These results suggested that deterioration of spermatozoa preserved in ETBS alone was delayed by supplementation with antioxidants.     

Freeze-thawing as the factor of spontaneous activation of spermatozoa motility in common carp (Cyprinus carpio L.)

In the present study, we investigated the possibility of spontaneous carp spermatozoa activation by freeze-thawing. To evaluate this, the parameters of spermatozoa motility percentage, velocity, ATP content level and fertility rate of sperm were used. The motility and velocity of spermatozoa activated by freeze-thawing were characterized by motile spermatozoa with a median value of 16% and a velocity of 98 μm/s. In addition, the motility and velocity of sperm from the thawed samples were significantly lower than in the control (median value of 100% for sperm motility and 175 μm/s for sperm velocity). Furthermore, a spontaneously activated spermatozoa motility terminated within five minutes post-thaw time. After freeze-thawing the ATP level significantly decreased with post-thaw time (46 nmol ATP/10⁹ and 10 nmol ATP/10⁹ at 25 s and 10 min after thawing, respectively). Fertility of spermatozoa was not significantly affected within 10 min post-thaw. On the other hand, the fertility of frozen-thawed sperm was significantly lower if compared to fresh sperm. We conclude that the freeze-thawing procedure spontaneously activated spermatozoa motility in common carp. However, this activation did not negatively affect the fertility of frozen-thawed sperm.     

Effect of dietary Vitamin E on the cloacal gland, foam and semen characteristics of male Japanese quail

This experiment was to investigate the effects of increasing the level of dietary Vitamin E (Vit. E) on cloacal gland size, foam production and semen characteristics of male Japanese quail (Coturnix coturnix Japonica). One hundred and eighty male Japanese quail chicks (day old) were randomly distributed to three dietary treatments for a period of 25 weeks. Each treatment comprised of three replicates each containing 20 chicks. The basal diet contained 15 IU Vit. E/kg and the two experimental diets were supplemented with 150 and 300 IU Vit. E/kg (diets T2 and T3, respectively). DL alpha-tocopherol acetate was used as the source of Vit. E. All chicks were provided feed and water ad libitum. Foam characteristics, in terms of frequency of foam discharge (24h), cloacal gland index and foam weight were significantly higher (P<0.05) in T2 group. Body weight, testes weight (left and right) and plasma testosterone concentrations did not differ significantly. Semen characteristics (semen volume, sperm motility, % live sperm, % hatchability and sperm concentrations) did not differ significantly (P>0.05). Percentages of abnormal and dead spermatozoa were significantly (P<0.05) lower and fertility was higher (P<0.05) in the T2 group. From this study, it can be concluded that moderate supplementation of dietary Vit. E may be beneficial for foam production, cloacal gland and improve the semen characteristics in male Japanese quail.