Comparison of spermatozoal movement and semen characteristics with fertility in stallions: 64 cases (1987-1988).

Information pertaining to evaluation of single ejaculates of semen and records for 2 consecutive breeding seasons were obtained. In all, data for 99 individual breeding seasons (n = 43 Standardbreds and 56 Thoroughbreds) were evaluated. Included in each semen evaluation was examination of semen characteristics and computer-aided analysis of spermatozoal movement characteristics. On the basis of the analysis of breeding records for 4,175 mares (7,017 estrous cycles), a per-estrous cycle fertility rate was calculated from data for 96 of the breeding seasons. Stallions with lower fertility than the mean overall season fertility had significantly (P less than 0.01) lower mean values for subjective appraisal of the percentage of motile and progressively motile spermatozoa and for percentage of morphologically normal spermatozoa. Lower mean values were obtained for computer-aided movement analysis of the percentage of motile and progressively motile spermatozoa, and for mean velocity of motile spermatozoa. Semen characteristics, including spermatozoal movement characteristics, and fertility were significantly (P less than 0.05) correlated for Thoroughbred and Standardbred stallions when analyzed individually and when data for both breeds were combined. Characteristics most highly correlated (P less than 0.01) with fertility data for both breeds combined were: subjective appraisal of the percentage of motile (r = 0.40) and progressively motile (r = 0.46) spermatozoa; percentage of morphologically normal spermatozoa (r = 0.36); and computer-aided analysis of percentage of motile spermatozoa (r = 0.34). However, on the basis of evaluation of a single ejaculate for each stallion, the variation in these characteristics only accounted for approximately 20% of the observed variation in fertility rate.     

Performing a complete canine semen evaluation in a small animal hospital

Normal dog semen ranges in volume from 1 to 30 mL per ejaculate and contains 300 million to 2 billion sperm, of which more than 70% are progressively motile and morphologically normal. Dog semen should contain fewer than 10,000 bacteria per mL; higher numbers are indication of an infection of the male reproductive tract and usually are associated with cytologic evidence of inflammation (neutrophils in semen sediment) and with decreased progressive motility and decreased numbers of morphologically normal sperm. Measurement of pH of seminal plasma of dogs with infection of the reproductive tract may provide information on determining the choice of antibiotic. Measurement of seminal plasma alkaline phosphatase in azoospermic dogs is an indicator of tubular patency to the level of the epididymides.     

Semen collection and evaluation

Conformation showing and a variety of dog sports are increasingly popular. Propagation of an excellent dog requires normal fertility. Semen collection and evaluation are essential skills for small animal practitioners who work with breeders and dog-sport enthusiasts. Semen collection can be performed on most dogs in the clinic setting with minimal standard equipment. Preparation of the collection room and presence of a teaser bitch increase the success of sample collection. Semen evaluation consists of several components. First, gross characteristics and sample volume are recorded. Percentage and quality of motility are evaluated immediately. The concentration of spermatozoa in the sample is determined and multiplied by the total sample volume to determine the total number of sperm in the ejaculate. Morphology of > or = 100 spermatozoa is evaluated and recorded to determine the percent normal spermatozoa Total normal sperm per ejaculate is then calculated. Additional tests are often performed. These include cytology of the second and third fractions, culture of the semen, and measurement of semen alkaline phosphatase. Knowledge of normal parameters for canine semen enables the practitioner to evaluate the results of these tests with confidence.     

Einflüsse von Prostatasekret und Verdünner auf die Spermienmotilität und ATP-Konzentration sowie die Aktivität der sauren und alkalischen Phosphatase von Beagle-Samen*

Influences of seminal plasma and extender on sperm motility, ATP-concen-tration, and the activity of acid and alkaline phosphatases of beagle dog semen. The percentage of progressively motile spermatozoa, the time of sperm survival, ATP-concentration, and the activity of the acid and alkaline phosphatases were determined in the semen of five healthy beagle dogs immediately after collection and after storage for 24 hours at +5°C. The sperm rich fraction was examined in the native state as well as after the addition of prostatic secretion and a Tris-eggyolk-medium, resp. The percentage of progressively motile spermatozoa was 64.4% in the fresh undiluted semen, 68.4% after the addition of prostatic secretion, and 74.8% after dilution with Tris-eggyolk-medium. It decreased within 24 hours to 31.6%, 20.4%, and 59.2%, resp. After 0 und 24 hours, resp., the sperm survival time in a coverslide preparation was     

In vitro assessment of semen for the prediction of fertility

The prediction of fertility is a primary goal in the field of reproductive medicine. The aim of the present paper is to describe the value of conventional and modern sperm analysis systems considering the process of fertilization. The classical assessment of motility and morphology enables the rough estimation of semen quality in order to select ejaculates for the use in artificial insemination. Recent methods for sperm diagnosis, such as fluorescent marking for the detection of sperm plasma membrane integrity, the hypoosmotic swelling test, and computer assisted semen analysis allow for the evaluation of a large number of spermatozoa and the assessment of sperm dynamics under in vitro-fertilization conditions. The oocyte penetration test investigates the ability of spermatozoa for capacitation, hyperactivation and acrosome reaction in vitro. The amount of specific seminal plasma proteins is related to fertility and thereby provides an additional semen evaluation method. For the use of a given semen test the specific in vitro condition has to be considered. In addition, the evaluated criteria relevant for the process of fertilization need to be defined. The combination of selected semen tests gives a higher accuracy for the prediction of fertilizing capacity compared with a single test.     

Relationship of Sperm Quality to Fertility after 4 Days of Cooled Storage of Equine Semen

This study evaluated measures of sperm quality in relation to fertility achieved with fresh semen or semen cooled and stored. Semen from 1 stallion was collected and processed to provide 3 treatments: group 1 received fresh semen; group 2 received cooled semen containing 50% seminal plasma (SP) stored for 4 days; and group 3 received cooled semen containing 50% SP stored for 1 day, then centrifuged and resuspended in fresh extender containing 10% SP on days 1 to 3. Inseminates were evaluated for sperm motion characteristics and the percentage of sperm with intact membranes (SMI). Mares (n = 34) in estrus were treated with an ovulation-inducing drug and inseminated with 100 million membrane-intact sperm on the following day. Pregnancy status was determined via transrectal ultrasonography 2 weeks after ovulation. The mean percentage of SMI was higher in group 1 (81%, initial) than in group 2 (74%, day 4) or group 3 (74%, day 4) (P < .05). The median percentages of total sperm motility differed among the groups (77%, 5%, 59% for groups 1, 2, and 3 respectively; P < .05). Median values for the percentages of progressively motile sperm and curvilinear velocity for group 1 (55%, 216 μm/s) and 3 (37%, 186 μm/s) were higher than for group 2 (1%, 73 μm/s) (P < .05). Pregnancy rates did not differ among groups (5 of 11, 45% in group 1; 5 of 11, 45% in group 2; and 7 of 12, 58%, in group 3; P = .77). These data suggest that, at least for this stallion, sperm membrane integrity may be a more valuable means of assessing potential fertility of cooled-stored semen than sperm motion characteristics.     

Cryopreservation of the semen collected by electroejaculation from the Hokkaido brown bear (Ursus arctos yesoensis)

Four adult Hokkaido brown bears were used as semen donors, and semen characteristics were examined before freezing and after thawing. A total of 10 electroejaculates were diluted with Tris-egg yolk extender and cooled to 4 degrees C over 90 min. Spermatozoa were equilibrated with 4.7% glycerol for 80 min. Semen packed in 0.25 ml plastic straws were frozen with liquid nitrogen vapor. Percentages (mean +/- SD) of motile and live sperm were 96+/-2 and 86.5+/-7.2% before freezing, and 43+/-5 and 67.4+/-3.9% after thawing, respectively. Although the number of progressively motile sperm after thawing varied among samples (1.8+/-1.2 x 10(8) cells/ejaculate), frozen semen in the present study might serve for artificial insemination.     

Seminal plasma components in camelids and comparisons with other species

Camelid semen is characterized by a highly viscous, low-volume ejaculate with a low concentration of spermatozoa that exhibit low progressive motility. The viscous seminal plasma is currently the major impediment to the development of assisted reproductive technologies (ARTs) in camelids. To advance ARTs such as sperm cryopreservation and artificial insemination in camelids, it is necessary to identify the cause of the viscosity and gain an understanding of the role of seminal plasma components on sperm function and fertility. Numerous compounds and proteins have been identified as mediators of sperm function and predictors of fertility in other livestock species, and understanding the importance of specific proteins has progressed the success of ARTs in these species. Current knowledge on the components of camelid seminal plasma is outlined, together with the implications of these components for the development of ARTs in camelids. The cause of semen viscosity, as well as proteins that are present in camelid seminal plasma, is described for the first time. Seminal plasma components are compared with those of other species to hypothesize their role in sperm function and fertility.     

The Relationship between Semen Quality and the Nonreturn Rate of Bulls

Contents
The value of routine evaluation of bull semen was analysed for 117 AI bulls placed in two studs. Data from semen analysis of a total of 1635 ejaculates was compared statistically with the nonreturn rates of the bulls. The semen parameters which were significantly correlated to nonreturn rates were the motility of the freshly collected ejaculates (p = 0.0140) and post-thaw motility (p = 0.0075). The total number of motile sperm in the inseminate ranged from 10.9 to 19.3 × 10⁶ and according to previous reports the effects of low motility should be fully 'compensated' when doses above 10 × 10⁶ sperm/dose are used for insemination. In conclusion, the motility of freshly collected semen does not appear to be 'compensation' and a low percentage of motile sperm in an ejaculate may indicate other dysfunctions of the population of motile cells. Furthermore, post-thaw motility appears to correlate significantly with nonreturn rates. The largest proportion of the variation was explained by the breed of the bull and stud (42.2% of the variation), whereas the two motility parameters explained 10% of the total variation in nonreturn rates. Objective and precise evaluation of sperm motility in combination with other semen traits are needed to improve breeding efficiency. Although microscopic evaluation of sperm motility correlates with nonreturn rates of bulls, the methods are subjectively assessed and inaccurate and therefore do not allow a satisfactory prediction of fertility.     

Fertility of Mares Inseminated With Frozen-Thawed Semen Processed by Single Layer Centrifugation Through a Colloid

The aim of this study was to determine whether there was an increase in pregnancy rates when frozen-thawed stallion semen was processed by single layer centrifugation (SLC) through a colloid before insemination. In addition, changes in semen parameters, including motility, were determined before and after SLC. Twenty light-horse mares (aged 3-16 years) and one Thoroughbred stallion (aged 16 years) having average fertility with fresh and cooled semen (>50% per cycle) and displaying a postthaw motility of >35% were used. Control mares were inseminated using 4- × 0.5-mL straws (200 × 10⁶/mL) of frozen-thawed semen. Treatment mares were inseminated with 4 × 0.5 mL of frozen-thawed semen after processing by SLC. Pregnancy rates were compared using Fisher exact test, and continuous parameters were evaluated by a Student t test. The pregnancy rates at day 14 were not different for the mares inseminated with control versus SLC-processed semen, despite the difference in sperm number (171 × 10⁶ ± 21, 59 × 10⁶ ± 25 progressively motile sperm). After frozen-thawed semen was processed by SLC, the percentage progressively motile sperm improved (P < .05), and SLC processing resulted in a 21.8% recovery of spermatozoa. In summary, centrifugation of frozen-thawed semen through a single layer of colloid increased the percentage of motile spermatozoa, but did not improve pregnancy rates after deep horn insemination.     

The Effects of Frequent Electroejaculation on the Semen Characteristics of a Captive Siberian Tiger (Panthera tigris altaica)

Artificial insemination (AI) can help to avoid inbreeding and genetic degeneration for sustaining genetically healthy populations of endangered species in captivity. Collection of a sufficient quantity of viable sperm is an essential first step in the AI process. In the present study, we examined the effects of frequent electroejaculation on semen characteristics in a Siberian tiger. We collected semen in all 17 trials during 6 breeding seasons (6 years). The mean number of sperm and the percentage of motile sperm were 294.3 ± 250.2×10⁶/ejaculate and 82.4 ± 11.4%, respectively. The number of motile sperm tended to increase during frequent electroejaculation in the same breeding season. Semen collection by electroejaculation can be performed effectively up to the fourth sequential ejaculate, which contained the most sperm in the study. In conclusion, frequent collection of sperm by electroejaculation from tigers may be effective for collection of a large number of motile sperm.     

Field data analysis of boar semen quality

This contribution provides an overview of approaches to correlate sow fertility data with boar semen quality characteristics. Large data sets of fertility data and ejaculate data are more suitable to analyse effects of semen quality characteristics on field fertility. Variation in fertility in sows is large. The effect of semen factors is relatively small and therefore impossible to find in smaller data sets. Large data sets allow for statistical corrections on both sow- and boar-related parameters. Remaining sow fertility variation can then be assigned to semen quality parameters, which is of huge interest to AI (artificial insemination) companies. Previous studies of Varkens KI Nederland to find the contribution to field fertility of (i) the number of sperm cells in an insemination dose, (ii) the sperm motility and morphological defects and (iii) the age of semen at the moment of insemination are discussed in context of the possibility to apply such knowledge to select boars on the basis of their sperm parameters for AI purposes.     

Efficacy of Low‐Dose Human Chorionic Gonadotropin Therapy in Dogs with Spermatogenic Dysfunction: A Preliminary Study

Human chorionic gonadotropin (hCG) is a glycoprotein used in the treatment of spermatogenic dysfunction. However, previous studies performed in dogs show that repeated administration of large doses of hCG produces antibodies against hCG. In this study, we examined the efficacy of low‐dose injections of hCG in four male dogs with spermatogenic dysfunction and low plasma testosterone (T) levels. We administered 100 IU hCG per animal, five times at 3‐day intervals, and evaluated the changes in semen quality and plasma T levels. The total number of sperm in ejaculate, the percentage of progressively motile sperm and the plasma T levels had increased by 3–5 weeks after the first injection of hCG in three of the four dogs, but were unchanged in the fourth dog. These findings indicate that temporary improvement of the semen quality of dogs with spermatogenic dysfunction and low plasma T levels is possible after five low‐dose injections of hCG.     

Function of cryopreserved horse semen is improved by optimized thawing rates

The use of cryopreserved semen for artificial insemination in equine breeding programs is increasing because of such benefits as reduction of injuries inherent in natural breeding. In the present study, semen straws from 1 ejaculate from each of 3 stallions predetermined to be poor freezers were thawed after long-term storage in liquid nitrogen (−196°C) at 3 different thaw rates (37°C for 30 seconds, 60°C for 8 seconds, 75°C for 7 seconds). Sperm samples were tested in triplicate for each stallion for motility in caffeine to detect all potentially motile sperm and sperm viability and capacitation status. The fast thaw rate significantly increased the percent of progressively motile sperm compared with the slow (P = .0113) and moderate (P = .0157) thaw rates. The fast thaw rate also improved sperm viability, as measured by the dual stain SYBR-14 and PI (P < .05). Similar improvements were achieved with semen from 3 stallions with average postthaw semen quality. Finally, the chlortetracycline (CTC) stain found that the fast thaw rate reduced the number of sperm undergoing premature acrosome reactions (P = .0329). In summary, thawing frozen equine sperm at 75°C for 7 seconds apparently caused less membrane damage on thawing resulting in enhanced postthaw motility and viability and reduced premature capacitation.     

Upregulation of CRISP-3 and kallikrein in stallion seminal plasma is associated with poor tolerance of cooled storage

For unknown reasons, stallion fertility and sperm longevity during cooled storage of semen vary markedly between individuals. Spermatozoa from individual stallions react differently to the presence, or the removal, of seminal plasma (SP). The aim was to evaluate differences in protein content in stallion seminal plasma with either a positive or a negative effect on sperm chromatin integrity during storage. Stallion semen samples from different ejaculate fractions were stored at 5°C for 24 hr. Sperm survival was assessed after storage using a sperm chromatin structure assay. Protein expression in SP with either positive or negative effects on sperm survival during storage was studied using two-dimensional differential gel electrophoresis and liquid chromatography–mass spectrometry. Lower sperm chromatin integrity was associated with upregulation of the proteins kallikrein, CRISP-3 and HSP-1, while higher chromatin integrity was associated with upregulation of TIMP-2. In the sperm-rich fractions, kallikrein and CRISP-3 differed significantly between SP samples with differing effects on sperm chromatin integrity. In the sperm-poor fractions, TIMP-2 and HSP-1 differed significantly between the two SP groups. Differences in the seminal plasma proteome are associated with sperm longevity during cooled storage.     

Age-related declines in ejaculate quality and sperm kinematics vary among strains of Japanese Quail (Coturnix japonica)

For successful breeding programs, it is important to quantify the useful period of a male's reproductive life and it is often done simply by measurement of semen quality. This information is lacking for Japanese quail so we tested whether there is a decline in ejaculate quality and sperm kinematics with age, and whether the decline varies among strains. Nine males (n = 9) from each of 5 strains (A, B, C, D and E) were subjected to 4 semen collections (n = 16 per male) at 8, 16, 26 and 36 weeks of age. Ejaculate volume, sperm concentration and total sperm per ejaculate were measured, and sperm kinematics were analysed using a Sperm Class Analyser (SCA^®). There was a significant effect of age for ejaculate volume, total sperm per ejaculate and per cent medium sperm. The effect of the interaction between age and strain was significant for percent progressive motile sperm, percent rapid sperm, velocity curvilinear, velocity straight line, velocity average path, linearity, straightness and beat cross frequency. Ejaculate volume peaked at Week 26 in all strains, while peak values for sperm concentration and total sperm per ejaculate were observed at Week 16 for most strains. There were declines in percent motile sperm, progressive motile sperm and rapid sperm, and in velocity curvilinear velocity, velocity straight line and velocity average path, by Week 16 for most strains. Linearity declined by Week 26 in some strains, and all strains showed a significant decline in beat cross frequency by that age. In conclusion, the ability of CASA to detect age-related changes in sperm kinematics makes it a valuable tool for identifying the best males and thus improving quail flock fertility. It is essential that breeders understand that age affects both sperm production and sperm kinematics, and that the changes vary with strain.     

Genetic Parameters for Semen Production Traits in Austrian Dual-Purpose Simmental Bulls

Genetic parameters were estimated for semen production traits collected in an Austrian AI centre in the years 2000-2004. In total, 12,746 ejaculates from 301 Austrian dual-purpose Simmental (Fleckvieh) AI bulls were examined considering different effects on ejaculate volume, sperm concentration, percentage of viable spermatozoa in the ejaculate, total spermatozoa per ejaculate and motility. The model for genetic parameter estimation included the fixed effects age of bull, collection interval, number of collections on collection day, bull handler, semen collector, year and month of collection, a random additive genetic component and a permanent environmental effect. Correlations between estimated breeding values for semen traits and male fertility from the routine evaluation were calculated. The fertility trait considered in the routine evaluation is non-return rate 90 for the first insemination. All semen production traits were moderately heritable. Heritabilities for volume, concentration, percentage of viable spermatozoa, total number of spermatozoa and motility were 0.18, 0.14, 0.10, 0.22 and 0.04, respectively. Correlations between breeding values for semen quality traits and routinely estimated breeding values for male fertility were low and ranged from 0.08 to 0.17 indicating that semen production traits are rather poor predictors of male fertility.     

Numbers of Sertoli cells, quantitative rates of sperm production, and the efficiency of spermatogenesis in relation to the daily sperm output and seminal quality of young beef bulls

Data from 34 yearling Hereford or Angus bulls were used to investigate relationships of testicular size, quantitative rates of sperm production, Sertoli cell numbers, numbers of germ cells supported per Sertoli cell, and the efficiency of spermatogenesis to daily sperm output and seminal quality. Two ejaculates were collected by electroejaculation from each bull on each of 2 days/week throughout the study. The percentage of progressively motile sperm and the percentage of morphologically normal sperm were determined from aliquots of fresh semen. Additional aliquots of semen were frozen in glass ampules or plastic straws and subsequently evaluated for postthaw motility and percentage of sperm with intact acrosomes. Sertoli cell numbers, the numbers of germ-cells per Sertoli cell, and the efficiency of spermatogenesis were unrelated to the quality of fresh or frozen semen (P greater than 0.05). In first ejaculates, the numbers of sperm and motile sperm were related (P less than 0.05) to testicular parenchymal weight (r = 0.38 and 0.50), daily sperm production (r = 0.45 and 0.53), and spermatids per gram of testicular parenchyma (r = 0.35 and 0.34). Testicular parenchymal weight and daily sperm production also were related to daily sperm output and to the average daily motile sperm output of these bulls (P less than 0.05), but could account for less than 25% of the variability in these end points among bulls.     

The Predictive Value of Porcine Seminal Parameters on Fertility Outcome under Commercial Conditions

The aim of this study was to address the question of whether differences in farrowing rate and litter size after the use of different ejaculates could be predicted using the standard semen parameters under commercial conditions. In this study, a total of 1818 sows were used to evaluate the fertility predictive value of different sperm parameters. Logistic regression analysis (univariate and multivariate) was used to correlate the dichotomous farrowing rate data to the sperm parameters. Linear regression was also used to determine the relationship between litter size and semen parameters (Pearson correlation and multiple regression). Receiver-operating curves (ROC) were used to determine the overall performance characteristics of each semen variable in the logistic regression model. Semen analysis, under commercial conditions, allows to identify ejaculates with very low fertility potential but the pre-selection of the samples, the high number of sperm per doses and the high quality of the semen used in artificial insemination (AI) programmes reduces the variability. Therefore, it is unlikely to detect fertility differences associated with seminal parameters.     

The Effect of Season on Spermatozoa Motility,Plasma Membrane and Acrosome Integrity in Fresh and Frozen–Thawed Semen from Xinong Saanen Bucks

The aim of this study was to evaluate whether the season of ejaculate collection influences seminal quality parameters of pre‐ and post‐freeze–thawing in Xinong Saanen bucks. Ejaculates were collected from eight bucks throughout the four seasons (spring, summer, autumn and winter) in a 12 months’ time period, identified in the Northern Hemisphere. Semen samples were evaluated by the combinations of conventional and Computer‐Assisted Sperm Analysis (CASA) when fresh and after frozen–thawed, respectively. The results clearly demonstrated that season of ejaculate collection influenced (p < 0.05) fresh semen quality. Highest semen quality was observed during autumn. On the contrary, undesirable indices (significantly lower, p < 0.05) were observed in winter as compared with the other remaining seasons. CASA has clearly shown the influences of seasonal variations on semen motility parameters. Furthermore, season of ejaculate collection was also found to influence sperm freezability. Semen characteristics after frozen–thawed followed a similar pattern with that of fresh ejaculate except in spring. The results revealed that sperm quality was higher (p < 0.01) in summer and autumn than in spring and winter. In conclusion, seasonal variation influences semen quality in Xinong Saanen bucks. In addition to summer and autumn, fresh ejaculates in spring can also be successfully used for AI. Sperm from ejaculates collected during summer and autumn are more suitable for cryopreservation. Hence, it is possible to increase the efficiency of goat breeding by manipulating the seasonal variations of semen quality for immediate AI and/or cryopreservation.