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.     

Summer heat stress alters the mRNA expression of selective-uptake and endocytotic receptors in bovine ovarian cells

Summer heat stress (HS) is a major factor in decreased reproductive performance in high-producing dairy cattle, possibly by affecting the steroidogenic capacity of ovarian follicles and ovarian follicular dynamics. In the present study, mRNA expression of cholesterol receptors was determined in bovine ovarian cells. Two endocytotic receptors (very-low-density lipoprotein receptor (VLDLr) and low-density lipoprotein receptor (LDLr)), and two selective-uptake receptors (scavenger receptor class B type 1 receptor (SRB1) and the lipoprotein-receptor-related protein 8 (LRP8)) were evaluated. Ovarian follicles in four diameter categories were evaluated from cows under non-heat stress (NHS) and HS conditions. As follicle size increased, expression of mRNA in NHS cows increased for the selective-uptake receptors, SRB1 and LRP8, and decreased (P<0.004) for the endocytotic receptors, LDLr and VLDLr. In contrast, in HS cows, mRNA expression did not significantly change (with increasing follicle diameter) for either receptor type. With increasing follicle diameter, cholesterol and fatty acid concentrations in the follicular fluid did not change in HS cows, whereas in NHS cows, cholesterol increased (P<0.008) and fatty acid decreased (P<0.0001). These changes paralleled those in the different lipoprotein fractions in the follicular fluid. In follicles from HS cows, the altered mRNA expression patterns for the endocytotic and selective-uptake receptors caused changes in the regulation of cholesterol supply at critical stages of folliculogenesis, which may play a role in the low turnover rates of ovarian follicles during the summer.     

The contribution of bulls and cows to the seasonal differences in the fertility of dairy cattle in Israel

Semen collected from dairy Israeli-Friesian bulls in the winter and summer was pelleted-frozen and used for inseminations in the collection season, in the other season, and after a 12-month storage period.The semen quality, as assessed by the percentage of ejaculates with initial poor motility and low sperm density and the percentage of spermatozoa surviving the freezing-thawing process, and those revealing acrosomal and tail abnormalities after thawing, was slightly and non-significantly lower in summer than in winter.The fertility of semen collected in both seasons was not affected by a 12-month storage period. The use of winter semen in summer did not improve the low fertility of cows in that season compared with those inseminated with summer semen.It appears from the results of this study that the summer decrease in the fertility of the inseminations is due to the adverse effect of this season on the fertility of the cows.     

Seasonal changes of semen quality and freezability in Franches-Montagnes stallions

The objective of this study was to investigate seasonal changes of semen quality parameters in Franches-Montagnes stallions and to compare the freezability of ejaculates collected in autumn and winter. Experiments were performed using 15 stallions from the National Stud Farm in Avenches (Switzerland). Ejaculates were collected and evaluated every month during 1 year as well as cryopreserved in autumn and winter (September to February). In fresh semen the gel-free volume, concentration, motility and morphology (normal sperm, major defects, vacuoles and acrosome defects) were evaluated and in frozen-thawed semen the motility as well as the viability (SYBR-14/PI) were performed. To analyse seasonal differences four periods of 3 months each were defined as autumn (September, October, November), winter (December, January, February), spring (March, April, May) and summer (June, July, August). During the 1-year experiment all fresh semen quality parameters demonstrated a clear seasonal and individual pattern. The gel-free volume was significantly (P<0.05) higher in spring and summer compared to autumn and winter while sperm concentration was significantly (P<0.05) lower in spring than at any other time of the year. Total sperm number was significantly (P<0.05) higher and sperm motility significantly (P<0.05) lower in summer than in other seasons. Regarding sperm morphology, normal sperm was significantly (P<0.05) higher in autumn than in winter and summer and major defects were lowest (P<0.05) in autumn. In frozen-thawed semen motility was significantly (P<0.05) improved in the ejaculates collected in autumn compared to winter, while viability showed no obvious differences. Our results clearly demonstrate that individual and seasonal differences occurred in semen quality of Franches-Montagnes stallions. Ejaculates collected in autumn (September, October, November) demonstrated good quality, especially regarding sperm morphology, and were more suitable for cryopreservation because of better motility in frozen-thawed semen collected during autumn than in winter.     

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.     

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.     

Seasonal changes in semen quality and freezability in the Warmblood stallion

The objective of this study was to investigate seasonal changes in stallion semen quality and to determine the best time for semen cryopreservation. Experiments were performed using 10 Warmblood stallions from the National Stud Farm in Avenches (Switzerland). Ejaculates were collected and frozen every other week during 1 year from January to December 1999. Volume, concentration, and motility, and the number of morphologically normal sperm and sperm with major defects (abnormal heads, acrosome defects, nuclear vacuoles, proximal droplets, abnormal midpieces) were evaluated. For all frozen-thawed semen samples motility as well as viability (SYBR-14/PI) was tested, and the hypoosmotic swelling test (HOS) was performed. To analyze seasonal differences 4 periods of 3 months each were defined: autumn (September, October, November), winter (December, January, February), spring (March, April, May) and summer (June, July, August). During the 1 year experiment all semen quality parameters showed a clear seasonal pattern. The volume, total sperm count and motility in fresh semen were significantly higher (P<0.05) in summer than in winter, while sperm concentration was significantly lower in summer compared to the other seasons. Regarding morphology, normal sperm was significantly lower (P<0.05) in summer than at any other time of the year and higher values (P<0.05) were found for major defects in summer than in spring and autumn. In frozen-thawed semen motility was significantly (P<0.05) improved in autumn when compared to spring and summer. Viability was lowest in summer and differed significantly (P<0.05) from other seasons. The HOS test revealed significantly more (P<0.05) membrane damaged spermatozoa in winter than in spring, summer and autumn. Our results demonstrate that in our climatic conditions clear seasonal differences occur in semen quality of fresh and frozen-thawed semen and that cryopreservation of stallion semen should preferably be performed in autumn.     

Detection of bovine papillomavirus type 2 DNA in commercial frozen semen of bulls (Bos taurus)

Papillomaviruses are found in epithelial lesions and are linked to different carcinogenic processes in humans and other animals. Although bovine papillomavirus (BPV) has been characterized as epitheliotropic, the presence of viral DNA has been detected in other sample types, including fresh semen. The aim of this study was to evaluate the presence of BPV DNA in spermatozoa and seminal plasma samples of commercial frozen semen taken from bulls (Bos taurus) and its effects on semen function. PCR assays were conducted with specific primers to detect BPV types 1-6 in 40 semen samples of dairy Gir bulls. The semen quality was assessed by the use of parameters such as motility, vigor, acrosomal integrity and DNA integrity. BPV-2 DNA was detected in all of the sperm cell samples and all the seminal samples; however BPV-1, 3, 4, 5 and 6 could not be detected. The presence of BPV DNA was apparently not a cause of reduced sperm function. This is the first record of BPV-2 DNA the commercial frozen semen taken from dairy Gir cattle by several companies that provide semen. Further studies are needed to assess the viability of the virus and the extent to which it can be spread through semen.     

Cholesterol concentration in seminal plasma as a predictive tool for quality semen evaluation

The aim of this study was to investigate the relationship between lipid composition of bovine serum and seminal plasma, seasonality, and semen quality. The experiment was carried out in two groups of Simmental breeding bulls: Group I (ages 2 to 4 yr) and Group II (ages 5 to 10 yr). Blood samples were collected from jugular vein, and bovine semen was sampled with an artificial vagina once per season. Serum concentrations of total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triacylglycerols, nonesterified fatty acids (NEFAs), and lipoprotein electrophoretic patterns were determined. Seminal plasma concentrations of total cholesterol, HDL-C, and LDL-C were assayed. Serum concentration of triacylglycerols in young bulls was significantly higher in winter compared with that in autumn, whereas serum NEFA concentration was significantly higher in autumn compared with that in other seasons. Serum concentration of total cholesterol, LDL-C, and LDL lipoproteins in older bulls was significantly higher in winter than in spring. Seminal plasma concentration of total cholesterol in young bulls was significantly higher in spring compared with that in summer, whereas in older bulls it was significantly higher in winter compared with that in autumn samples. Sperm volume of both groups was significantly higher in summer compared with that in autumn and winter. Sperm motility in young bulls was lowest in summer and differed significantly from the values recorded in other seasons. The changes observed in seminal plasma cholesterol concentration were associated with extracellular lipid use and appeared to be applicable as a biochemical marker of sperm quality.     

Seminal plasma damages sperm during cryopreservation, but its presence during thawing improves semen quality and conception rates in boars with poor post-thaw semen quality

To determine the effects of seminal plasma during and after cyopreservation on post-thaw sperm functions in semen from poor freezability boars, seminal plasma was removed immediately after collection, and sperm was subjected to cooling and freezing. Removal of seminal plasma did not significantly affect post-thaw sperm motility in good freezability boars; however, in boars with poor freezability, it increased post-thaw motility relative to control sperm cooled with seminal plasma (64.5+/-3.4% vs. 30.9+/-3.1%, P<0.01). Freezing sperm without seminal plasma increased both loss of the acrosome cap (37.5+/-1.6% vs. 18.4+/-2.8%, P<0.01) and expression of a 15 kDa tyrosine-phosphorylated protein (capacitation marker) in thawed sperm relative to controls; the addition of 10% (v/v) seminal plasma to the thawing solution significantly suppressed both changes and increased conception rate to AI (70% vs. 9% in the control group, P<0.05). In conclusion, our novel cryopreservation and thawing method increased the success of AI with frozen-thawed porcine semen, particularly from boars with poor post-thaw semen quality.     

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.     

Physiological response and semen quality of rabbit bucks supplemented with Moringa leaves ethanolic extract during summer season

Exposure of rabbit bucks to summer heat stress reduces their homeostasis and semen quality leading to a temporal subfertility. The potentiality of ethanolic extract of Moringa oleifera leaves (M. oleifera ethanolic extract (MLEE)) to reduce negative impacts of heat stress on physiological and semen quality traits was investigated. A total of 28 adult V-line rabbit bucks were randomly distributed among four experimental groups of seven rabbits each. The first group received water (placebo) and served as a control (M0). The other three groups were given orally MLEE at levels of 50 (M50), 100 (M100) and 150 (M150) mg/kg BW every other day for 12 consecutive weeks during the summer season. Chemical constituents of MLEE were detected by gas chromatography/MS. During the experimental period, ambient temperature and relative humidity were recorded daily and were used to estimate temperature and humidity index. Feed intake, BW, rectal temperature were recorded and blood serum biochemical attributes were determined. Semen samples were collected weekly and were analyzed for semen quality traits. Results showed that MLEE contained high percentages of long-chain fatty acids and antioxidant agents. Feed intake and BW were not affected significantly by the treatment, however rectal temperature was decreased significantly by 0.42°C, 0.24°C and 0.40°C in the M50, M100 and M150 groups, respectively, compared with the M0 group. Treatment with 50 mg/kg BW increased concentration of serum albumin (115%; P<0.05), total antioxidant capacity (132%; P<0.05) and testosterone (160%; P=0.098) as well as seminal plasma initial fructose (127%; P=0.092) compared with the control group. Compared with the control, MLEE supplementation with 50, 100 and 150 mg/kg BW increased significantly sperm concentration by 118%, 151% and 158%, sperm progressive motility by 117%, 120% and 118%, sperm viability by 129%, 137% and 127%, sperm normal morphology by 114%, 113% and 114%, intact acrosome sperm by 109% (on average) and sperm with integrated cell membrane by 109%, 123% and 114%, respectively. In conclusion, MLEE supplementation at a level of 50 mg/kg BW could be effectively used to improve heat tolerance, oxidative status and semen quality of rabbit bucks during summer season.     

Lipid profiles of sperm and seminal plasma from boars having normal or low sperm motility

Sperm plasma membrane lipids have an important role to play in determining membrane fluidity and sperm motility. The objective of the present study was to determine whether there are differences in the lipid and fatty acid (FA) composition of boar sperm and seminal plasma in the ejaculates of boars having different sperm motilities. Semen was collected from two groups of boars having normal (> 60%; n = 53) or low (< 60%; n = 53) motility sperm and the semen was evaluated for motility, morphology and vitality. The semen was then centrifuged to separate the sperm from the seminal plasma and both were kept at −20 °C until analyzed for lipid content and FA profile by gas chromatography. Total antioxidant status (TAS) of seminal plasma was determined using a commercial kit. There were differences (P ≤ 0.05) in sperm total lipids, cholesterol, saturated fatty acids (SFA), phospholipids, n-3 polyunsaturated fatty acids (PUFA), docosahexaenoic acid (DHA) and the ratio of n-6:n-3 PUFA between boars with normal and low motility sperm. Total lipids, cholesterol, phospholipids, PUFA, DHA and n-3 PUFA were positively correlated with sperm motility, viability, normal morphology and normal plasma membrane. In contrast, SFA and the ratio of n-6: n-3 PUFA were negatively correlated (P ≤ 0.05) with sperm motility, viability, normal morphology and normal plasma membranes. The TAS of seminal plasma from boars having normal motility sperm was higher (P ≤ 0.05) than that of boars having low motility sperm and TAS was positively correlated (P = 0.0001) with sperm motility, viability, normal morphology and normal plasma membranes. In summary, differences in sperm motility were related to n-3 PUFA content in the sperm plasma membrane and extracellular antioxidants in seminal plasma which protect sperm plasma membranes from lipid peroxidation during periods of oxidative stress.     

Seasonal emission of seminal coagulum and in vivo sperm dynamics in the black-handed spider monkey (Ateles geoffroyi)

The ejaculate of diverse primate species consists of two portions, liquid and solid; the latter, known as the seminal coagulum, is thought to sequester large numbers of sperm. In the black-handed spider monkey (Ateles geoffroyi), ejaculates collected by electroejaculation did not always contain seminal coagulum. The objective of the present study was to determine seasonal emission of seminal coagulum and in vivo sperm dynamics in the black-handed spider monkey. Seminal coagulum emission was related to season; it was more frequent in the dry season, coincident with maximal female fertility. Sperm concentration was higher (P = 0.02) in the dry season (dry vs. rainy season: 137.9 +/- 15.7 sperm/mL vs. 82.56 +/- 14.7 x1 0(6) sperm/mL; mean +/- S.E.M.) but also in ejaculates (collected during the rainy season) that had seminal coagulum (coagulum vs. no coagulum: 140.0 +/- 29.3 sperm/mL vs. 31.2+/-0.1 x 10(6) sperm/mL, P<0.001). In semen samples collected from the uterus after AI, the percentage of linearly motile sperm was higher during the dry season (dry vs. rainy: 9.1+/-2.1% vs. 5.9+/-2.5%), as well as whenever coagulum was present (coagulum vs. no coagulum: 13.0+/-3.2% vs. 2.0+/-0.9%, P<0.001).     

Endogenous prostaglandin F2α concentrations in bovine whole semen, seminal plasma, and extended semen

A series of experiments were conducted to quantify PGF_2α in bovine semen, seminal plasma, and extended semen, and to determine if PGF_2α was synthesized or released during extension of bovine semen. Concentrations of PGF_2α were measured in paired samples of whole and extended semen from beef and dairy bulls. Concentrations of PGF_2α did not differ between beef and dairy (mean ± SEM, 273.8 ± 42.8 vs. 210.3 ± 18.5 pg/mL, respectively; P = 0.12), but tended (P = 0.08) to be greater for whole compared with extended semen (255.5 ± 29.8 vs. 194.5 ± 17.0 pg/mL). Whole semen was extended at eight dilution rates (regardless of initial sperm concentration), using a diluent consisting of two fractions. Samples collected after each dilution step resulted in four subsamples. Concentrations of PGF_2α in subsamples decreased (P < 0.001) at higher dilution rates and later steps of extension. Subsequently, whole semen and seminal plasma were extended at three dilution rates. Initial PGF_2α concentration was greater (P < 0.001) for whole semen compared with seminal plasma. During extension, PGF_2α synthesis or release resulted in less disparity, but the amount synthesized or released was greater (P = 0.03) for semen compared with seminal plasma. We concluded that synthesis or release of PGF_2α during extension resulted in concentrations similar to whole semen.     

Season of ejaculate collection influences the freezability of boar spermatozoa

The aim of this retrospective study was to evaluate whether the season of ejaculate collection influences the freezability of porcine sperm. A total of 434 ejaculates were collected from boars of six different breeds over three years (2008–2011) and throughout the four seasons of the year identified in the northern hemisphere (winter, spring, summer and autumn). The ejaculates were cryopreserved using a standard 0.5 mL straw freezing protocol. Sperm quality was assessed before (fresh semen samples kept 24 h at 17 °C) and after freezing and thawing (at 30 and 150 min post-thawing in semen samples kept in a water bath at 37 °C), according to the percentages of total motility, as assessed by the CASA system, and viability, as assessed by flow cytometry after staining with SYBR-14, PI and PE-PNA. The data, in percentages, on sperm motility and viability after freezing and thawing were obtained at each evaluation time (recovered) and were normalized to the values before freezing (normalized). The season of ejaculate collection influenced (P < 0.01) sperm quality before freezing and after thawing (recovered and normalized), irrespective of the breed of boar. Sperm quality was lower in summer, both in terms of motility and viability, and in autumn, in terms of motility, than in winter and spring. Seasonality in the normalized data indicates that the season of ejaculate collection influences sperm freezability, regardless of the season’s influence on sperm quality before freezing. Consequently, the spermatozoa from ejaculates collected during summer and, to a lesser extent, also in autumn, are more sensitive to cryopreservation than those from ejaculates collected during winter and spring.     

Effects of age and environmental factors on semen production and semen quality of Austrian Simmental bulls

More than 90% of the breeding stock of Austrian dual purpose Simmental cows is artificially inseminated. Knowledge of factors affecting sperm production and semen quality is of importance with regard to reproductive efficiency and thus genetic improvement as well as for the productivity and profitability of AI centers. Hence, semen data from two Austrian AI centres collected in the years 2000 and 2001 were evaluated. In total, 3625 and 3654 ejaculates from 147 and 127 AI bulls, respectively, were analysed regarding ejaculate volume, sperm concentration, percentage of viable spermatozoa in the ejaculate, total spermatozoa per ejaculate and motility. Effects accounted for were the bull (random), age of bull, collection interval, number of collection on collection day, bull handler, semen collector, temperature on day of semen collection, in the course of epididymal maturation (average temperature of days 1-11 before collection) and during spermatogenesis (average temperature of days 12-65 before collection). Age of bull significantly affected all traits (P<0.01 to P<0.001) except motility score in center 2. Ejaculate volume and total number of spermatozoa increased with age of bull while sperm concentration was lower in higher age classes (center 1). The collection team was also found to significantly influence semen quality traits. With increasing collection interval ejaculate volume and total number of spermatozoa increased significantly (P<0.05 to P<0.001) while collection intervals between 4-9 days and 1-6 days were superior with regard to sperm concentration and percentage of viable spermatozoa, respectively (P<0.10 to P<0.001). First ejaculates were superior with respect to ejaculate volumes, sperm concentrations and total number of spermatozoa per ejaculate (P<0.001). Temperature, either on day of semen collection or during epididymal maturation or spermatogenesis, had important but inconsistent effects on semen production and sperm quality. Overall, however, ambient temperatures in the range of 5-15 degrees C were found to be optimal for semen production.     

Collection method, season and individual variation on seminal characteristics in the llama (Lama glama)

The objective of the present study was to evaluate the effect of semen collection method (electroejaculation "EE" as compared with the artificial vagina "AV"), the season (summer versus winter) and the male used on macroscopic and microscopic characteristics of ejaculates in llamas. A total of 110 ejaculates were collected from six males and 92 of them were analyzed. Ejaculate volume, concentration, total sperm and the following sperm characteristics were studied: motility, membrane function (HOS test), membrane integrity (CFDA/PI fluorochromes) and morphology. A mixed linear model, that considered season and collection method as the fixed variables and the male as the random variable, was used for the statistical analysis. Variability was found between males (p相似文献   

Intrafollicular insemination for the treatment of infertility in the dairy cow

Intrafollicular insemination (IFI) is based on direct introduction of a sperm suspension into a preovulatory follicle. To our knowledge, the first cases of IFI resulting in pregnancy of the cow are reported here. The experiment was performed on a dairy herd with low fertility during the warm season of the year. Following the sequence of estrus detection, one in three inseminations was intrafollicular (n = 17), whereas deposition of semen was performed into the uterine body (IUI) in the remaining cows (n = 33). Approximately 0.06 ml of a seminal dose containing five million spermatozoa (one quarter of a commercial seminal dose in a 0.25 ml French straw) was injected trans-vaginally into the preovulatory follicle in the IFI group. Four (23.5%) and 3 (9%) cows of the IFI and IUI groups, respectively, became pregnant. On days 8-11 post-insemination, ovulation failure was registered in 3 (18%) and in 4 (12%) cows of the IFI and IUI groups, respectively. Our results suggest that IFI could be used as an alternative procedure to the usual deposition of semen into the uterus in cows of low fertility.     

Influence of vedaprofen (Quadrisol) on quality and freezability of stallion semen

The objective of this study was to evaluate the effect of the non-steroidal anti-inflammatory drug (NSAID) vedaprofen (Quadrisol) on quality and freezability of stallion semen. Experiments were performed using 22 Franches Montagnes stallions from the National Stud in Avenches (Switzerland) randomly divided into a control and test group. Vedaprofen was given orally to all stallions of the test group at the recommended therapeutic dose (initial dose of 2mg/kg followed by 1mg/kg body weight every 12h) for 14 days. Control animals received the same amount of carrier substance. During treatment, blood samples of five stallions in both test and control group were collected for PGF(2 alpha)-metabolite (PG-metabolite) determination. Ejaculates from all stallions were collected and cryopreserved weekly for 14 weeks from September to December. Concentrations of PG-metabolite, PGF and PGE were measured in the seminal plasma of ejaculates collected 2 weeks before, during and 2 weeks after treatment. In fresh semen the volume, concentration, motility and number of normal sperm and sperm with major defects (acrosome defects, abnormal heads, nuclear vacuoles, proximal droplets, midpiece defects) were evaluated. In frozen-thawed semen samples motility as well as viability (SYBR-14/PI) were tested and the hypoosmotic swelling test (HOS) was performed. Results demonstrate that vedaprofen had no effect on blood plasma concentration of PG-metabolite but significantly inhibited both, PGF and PGE concentrations in seminal plasma. Furthermore, all quality parameters in fresh and frozen-thawed semen were not affected by vedaprofen treatment but the time of semen collection had a significant (P<0.05) effect on motility, normal sperm and sperm with nuclear vacuoles in fresh semen.