Genetic and non-genetic parameters of several characteristics of production and semen quality in young bucks

The objective of this study was to evaluate genetic and non-genetic factors influencing characteristics of young buck semen production using a multivariate model that takes into account the longitudinal structure of data. Data were collected from 1989 to 2002 at two French A.I. centres. The data corresponded to 13151 and 9206 ejaculates of 758 Alpine and 535 Saanen bucks respectively, collected at the beginning of the first breeding season (September-December). The semen volume, the total number of spermatozoa, the concentration, the motility score of spermatozoa after freezing and the percentage of motile spermatozoa after freezing were registered for each ejaculate. Within-breed heritabilities and repeatabilities were estimated using a multivariate animal model using a power spatial covariance structure for environmental effect. For all characteristics and the two breeds, the main source of variation was the year-month interaction that interacted with the centre. We observed a decrease in years of motility score after freezing. Age and frequency of collection had a significant effect on semen volume and number of spermatozoa for both breeds, and on concentration of spermatozoa for the Alpine breed. No effect of these factors was found on the characteristics observed after freezing. Heritabilities for concentration, number of spermatozoa, semen volume, motility score after freezing and percentage of motile spermatozoa after freezing per ejaculate were respectively, 0.32, 0.15, 0.25, 0.12 and 0.05 for the Saanen breed and 0.34, 0.25, 0.29, 0.17 and 0.03 for the Alpine breed. Genetic correlations between volume and number of spermatozoa were respectively, 0.74 for the Alpine breed and 0.86 for the Saanen breed. Further study is required to compare the semen characteristics of young bucks with their mature production.     

Assessment of goat semen freezability according to the spermatozoa characteristics from fresh and frozen samples

A total of 110 ejaculates were assessed in order to determine the influence of the physical parameters of goat semen on post-thaw motility and acrosome integrity. Sperm ejaculate characteristics, sperm motility, morphology and acrosome integrity were assessed in fresh and frozen samples by the sperm class analyzer (SCA) and Spermac staining technique. A decrease in acrosome integrity and sperm motility was found after thawing (P<0.01). Six semen parameters assessed before freezing were identified as predictors of sperm freezability (P<0.01). The percentage of morphological abnormalities (R=0.856) and motile sperm cells (R=0.655) in fresh semen are the best predictors to know the total post-thaw variability parameters.     

Genetic and environmental effects on semen traits in Lacaune and Manech tête rousse AI rams (Open Access publication)

Data from 51 107 and 11 839 ejaculates collected on rams of the "Lacaune" and "Manech tête rousse" breeds, respectively, were analysed to determine environmental and genetic factors affecting semen production traits (ejaculate volume, semen concentration, number of spermatozoa and motility) in young (≤1 year) and adult (≥2 years) rams. Fixed effects and variance components were estimated using multiple trait animal models within each breed. For all traits, the main environmental effects identified were year, season, number of ejaculations, daily variation, interval from previous to current collection and age. Heritability estimates were moderate for volume, concentration and number of spermatozoa (0.12 to 0.33) and lower for motility (0.02 to 0.14). Genetic correlations between ages differed from 1 for all traits (0.14 to 0.90), indicating that semen characteristics corresponded to different traits in young and adult rams. Genetic and phenotypic correlations among traits within age category were globally similar for the different breeds and categories of animals.     

The effect of straw size, freezing rate and thawing rate upon post-thaw quality of dog semen

Optimal freeze-thaw processes for dog semen will yield a maximal number of insemination doses from an ejaculate. The objectives of this study were to compare the effects of two straw sizes (0.25- and 0.5-mL French), two freezing rates (straws suspended 3.5 and 8 cm above liquid nitrogen) and two thawing rates (in water at 37 and 70 degrees C) upon post-thaw quality of dog semen, and to determine the best treatment combination. Quality was expressed in terms of the percentage progressively motile sperm 5 and 60 min after thawing and the percentage of abnormal acrosomes 5 min after thawing. One ejaculate from each of eight dogs was frozen. Two straws from each ejaculate were exposed to each of the eight treatment combinations. Data were analyzed by means of a repeated measures factorial analysis of variance and means compared using Bonferroni's test. Dog affected each response variable (P < 0.01). Neither straw size, nor freezing rate, nor thawing rate affected motility 5 min after thawing (P > 0.05). Half-milliliter straws resulted in 5.7% more progressively motile sperm 60 min after thawing and 6.5% fewer abnormal acrosomes than 0.25-mL straws (P < 0.05, n = 64). The percentage progressively motile sperm 60 min after thawing tended to be higher for semen thawed at 70 degrees C compared to 37 degrees C (P < 0.06, n = 64). Semen thawed in water at 70 degrees C had 6.6% fewer abnormal acrosomes than semen thawed in water at 37 degrees C (P < 0.05, n = 64). Freezing rate interacted with thawing rate (P < 0.05) in their effects upon acrosomal morphology and freezing 8 cm above liquid nitrogen and thawing in water at 70 degrees C was best. Dog semen should be frozen in 0.5-mL straws, 8 cm above liquid nitrogen and thawed in water at 70 degrees C.     

Effect of filtration on post-thaw quality of bull semen

Semen from 4 Holstein bulls was diluted in 4 different extenders, filtered with Sephadex ion-exchange column, and frozen in liquid nitrogen. Sperm motility, progressive motility, path velocity, progressive velocity and the percentage of normal acrosomes of filtered and nonfiltered semen were recorded before and after freezing. Semen characteristics were significantly influenced by extender, filtration and freezing. Before and after freezing, motility measurements and the percentage of normal acrosomes were higher (P < 0.001) in filtered than in nonfiltered spermatozoa. Post-thaw recovery rate of motile spermatozoa was higher in filtered semen than nonfiltered (68 vs 39%, P < 0.0001). The reduction in motility, progressive motility and the percentage of normal acrosomes during freezing and thawing processes were significantly lower (P < 0.0001) in filtered semen (34, 34 and 4%, respectively) than nonfiltered (59, 54 and 15%, respectively). Post-thaw viability of spermatozoa was significantly affected by extender, filtration and time (P < 0.0001). Immediate (0 h) post-thaw motility of nonfiltered semen (29%) was similar to 4-h post-thaw motility of filtered semen (25%; P > 0.05). In conclusion, bull spermatozoa recovered by Sephadex ion-exchange filtration showed better post-thaw viability.     

Comparison of TNC and standard extender on post-thaw quality and in vivo fertility of Thai native chicken sperm

Semen extender has a vital role in preservation of sperm cells properties in terms of sperm viability, motility, acrosome integrity, and mitochondrial membrane potential. The objective of the present study was to evaluate a new extender, known as Thai native chicken (TNC) extender compared to BHSV-based and modified Sasaki extenders for freezing chicken semen. Semen from Thai native roosters was collected, pooled, and randomly divided into three groups. Semen was frozen with a simple freezing method using nitrogen vapor and dimethylformamide. In the first experiment, post-thaw motion parameters, viability, acrosome integrity, mitochondrial function, and lipid peroxidation levels were analyzed using computer-assisted sperm analysis, propidium iodide, fluorescein isothiocyanate-conjugate peanut agglutinin, JC-1, and the thiobarbituric acid reaction. Results showed that the type of extender had no effect on the percentage of total motile and curvilinear velocity. The percentage of progressive motile, straight-line velocity, and average path velocity of post-thawed semen were significantly lower in TNC compared to the modified Sasaki extender. However, the percentages of post-thawed acrosome integrity and active mitochondria were significantly higher in TNC extender (P < 0.05). For the second experiment, semen was thawed by using each of extenders thereafter, was inseminated to 48-layer breeder hens to determine the fertility rate. Among the three extenders used, the highest fertility rate was found in TNC extender. In conclusion, TNC extender can be recommended as an appropriate and useful cryopreservation media for native chicken semen since it maintains the quality of rooster semen and fertility after freezing and thawing process.     

Semen production and fertility issues related to differences in genetic lines of boars

Boars have been reported to differ in various characteristics important to productivity of AI units. These aspects include sensitivity to seasonal infertility, ejaculate volume x age interactions, optimum collection frequency, discard (trash) rate of ejaculates, age of puberty, libido and trainability, and semen shelf life. In addition to differences among individual boars, there were also identifiable differences among breeds, between purebreds and crossbreds, and between lines. The percentage of ejaculates discarded varied among seasons and among breeds within season, with maternal purebred lines being the most sensitive to warmer temperatures. These same lines were also slower to mature and produced fewer sperm per ejaculate. Breeds differed in the sustainability of motility in stored extended semen, and in farrowing rates. Boar centers need to factor breed differences into their decision-making processes to ensure adequate boar power and customer satisfaction.     

Characteristics and seasonal variations in the semen of Alpine, Saanen and Damascus goat bucks born and raised in Greece

Twenty-three purebred Alpine (n=8), Saanen (n=7) and Damascus (n=8) goat bucks raised at the Institute of Reproduction and Artificial Insemination in Thessaloniki, Greece (40 degrees 37 minutes N, 22 degrees 58 minutes E and altitude 32 m above sea level), were used to study the effect of photoperiod on semen production. Samples were collected with an artificial vagina and examined immediately after collection. In spite of the variation in nearly all semen characteristics studied among the 3 breeds of bucks, there was significant seasonal variation in both semen quantity (volume, concentration and total number of spermatozoa per ejaculate) and quality (percentage of motile spermatozoa, percentage of abnormal spermatozoa and rate of progressive motility). The best semen was produced during the breeding season (late summer and autumn). However, the magnitude of these seasonal effects was not sufficient to prevent bucks from being used for breeding throughout the year. Nevertheless, individual differences in the semen quantity and quality among bucks within a breed make individual evaluation of semen necessary to select the most fertile males for breeding.     

Supplementing rooster sperm with Cholesterol-Loaded-Cyclodextrin improves fertility after cryopreservation

Little is known about the effects of Cholesterol-Loaded Cyclodextrin (CLC) on post-thaw semen quality in chicken. The aim of the present study is to investigate the efficacy of CLC levels (0, 1, 2 and 3 mg/mL Schramm diluent) on post-thawed semen quality and fertility in two breeds of chicken Pradu Hang Dum (native chicken) and Rhode Island Red. Semen samples of each breed were pooled, divided into 4 aliquots and diluted with Schramm diluents, cooled to 5 °C when DMF was added (6% of final volume). Semen straws were subjected to cryopreservation using the liquid nitrogen vapor method. Post-thawed sperm motility, viability, acrosome integrity, mitochondrial function, and the Malondialdehyde (MDA) level were determined. The fertility of frozen semen was tested by inseminating laying hens. Post-thaw motility between Pradu Hang Dum and Rhode Island Red was no different; but Rhode Island Red had a higher semen viability and live cell intact acrosomes than Pradu Hang Dum (P < 0.05). The percentage of high functioning mitochondria in the Pradu Hang Dum was higher than the Rhode Island Red. CLC at 2 and 3 mg/mL supplementation was associated with improved viability of frozen semen; that is, acrosome integrity and mitochondrial function (P < 0.01), albeit having no effect on MDA levels. The sperm with 1 mg/mL CLC yielded a significantly better fertility (P < 0.01). CLC (1 mg/mL) improved the quality of frozen rooster semen. There was no interaction among breeds and CLC on post-thaw semen quality and fertility.     

The expression of the aryl hydrocarbon receptor in reproductive and neuroendocrine tissues during the estrous cycle in the pig

The sprouted wheat (SW) contains the 6-methoxy-2-benzoxazolinone (6-MBOA), a phenol compound that stimulates reproduction in certain small wild herbivorous mammals. The objective of the present study was to evaluate the effect of short-term supplemental dietary SW on libido, semen and sperm characteristics of rabbit bucks. Five-month old New Zealand White pubertal rabbits (n=18) were randomly allocated to one of two treatments: supplementation or not (control) supplemented with SW. The experimental design was completely random with nine replications, experimental unit was one buck. Semen collection for each male was conducted once a week with two ejaculations during 20 weeks. The SW was given during four consecutive days prior to each semen collection. Analysis of variance was under a mixed model: treatment, ejaculate number and season were fixed and rabbit random effects. There was no effect of treatment (P>0.05) on reaction time, gel presence, volume, pH, sperm motility, sperm number per ml and sperm number per ejaculate. The percentage of normal alive spermatozoa was 13.5% greater in SW-supplemented bucks than in the control and the percentage of abnormal alive spermatozoa was 44.1% greater in the control than in the SW-supplemented bucks. The morphology of dead spermatozoa, integrity of acrosome, number of normal alive motile sperm and semen doses per ejaculate were not influenced (P>0.05) by SW supplementation. The proportion of presence of gel and semen volume in the first ejaculate was greater than the second ejaculate (+140% and +56.4%). However, the semen quality in the latter was greater (P=0.0001) than the former in terms of an increase in motility (+29.7%). Reproductive traits were more desirable (P<0.05) in winter than autumn. Dietary wilted SW as a source of biological 6-MBOA enhanced sperm characteristics in terms of a greater percentage of normal alive and lesser percentage of abnormal alive spermatozoa but did not affect the number of normal motile live sperm and suitable semen doses in rabbit bucks in autumn and winter.     

Effect of dialysis or centrifugation on post-thaw motility and fertility of Santa Gertrudis bull semen collected by electroejaculation

Electroejaculated semen from Santa Gertrudis bulls was used to study the effect of centrifugation (600 x g for 5 min) or dialysis [molecular weight cutoff <14,000 Daltons (Da)] on post-thaw motility and on fertility in beef cattle. Analysis of post-thaw motility showed that the main effects (bulls and semen treatment) were significant (P<0.05). Dialysis significantly improved post-thaw motility in four of seven bulls. Initial volume of seminal plasma in the ejaculate was negatively correlated to post-thaw motility (r = -0.73). No significant improvement in post-thaw motility was observed for bulls with high volumes of ejaculate. In a fertility trial, calving rates of heifers synchronized with PGF(2)alpha and inseminated at 72 and 96 h after the second PGF(2)alpha injection with dialyzed or commercial semen were not statistically different (P>0.05; 54.4% vs 55.4%). These results show that dialysis could be used to improve post-thaw motility of electroejaculated bull semen without altering its fertilizing capacity. However, a high initial volume of seminal plasma seems to have a deleterious effect on sperm freezability that cannot be reversed by dialysis.     

Effect of semen collection practices on sperm characteristics before and after storage and on fertility of stallions

This study analyzed effects of different methods and intervals of semen collection on the quantity and quality of fresh, cool-stored, and frozen-thawed sperm and fertility of AI stallions. In Experiment 1, ejaculates were obtained from six stallions (72 ejaculates per stallion) using fractionated versus non-fractionated semen collection techniques. Initial sperm quality of the first three jets of the ejaculate was not different from that of total ejaculates. Centrifugation of sperm-rich fractions before freezing improved post-thaw motility and sperm membrane integrity when compared to non-centrifuged sperm-rich fractions or non-fractionated centrifuged ejaculates (P<0.05). In Experiment 2, semen from four stallions (60-70 ejaculates per stallion) was collected either once daily or two times 1h apart every 48 h. The first ejaculates of double collections had significantly higher sperm concentrations, percentages of progressively motile sperm (PMS) after storage for 24h at 5 degrees C and lower percentages of midpiece alterations than single daily ejaculates. Semen collected once daily showed significantly lower values of live sperm after freezing and thawing than the first ejaculate of two ejaculates collected 1h apart every 48 h. In Experiment 3, semen was collected from 36 stallions (> or =12 ejaculates per stallion) during the non-breeding season and the time to ejaculation and the number of mounts was recorded. When time to ejaculation and the number of mounts increased, volume and total sperm count (TSC) also increased (P<0.05), whereas a decrease was observed in sperm concentration, percentage of PMS after storage for 24 h at 5 degrees C, percentage of membrane-intact sperm in fresh semen (P<0.05) as well as motility and percentage of membrane-intact sperm of frozen-thawed sperm (P<0.05). In Experiment 4, AI data of 71 stallions were retrospectively analyzed for the effect of number of mounts per ejaculation and frequency, time interval of semen collections on pregnancy, and foaling rates (FRs) of mares. Semen volume increased, but sperm concentration and percentage of PMS after 24-h cool-storage decreased with increasing number of mounts on the phantom (P<0.05). A statistically significant inter-relationship was demonstrated between frequency and interval of semen collection and FR. Mares inseminated with stallions from which semen was collected frequently (> or =1 on an average per day) showed significantly higher FRs than mares inseminated with semen from stallions with a daily collection frequency of 0.5-1 or <0.5. FR of mares inseminated with stallions having 0.5-1 days between semen collections was significantly better than FR of mares that were inseminated with stallions having semen collection intervals of 1-1.5 days or >2.5 days.     

Effect of the collection method on semen characteristics of Zebu and European type cattle in the tropics

The effect of the collection method on the characteristics of fresh semen and the recovery of spermatozoa after thawing was studied in 30 Zebu bulls (Bos indicus ) and in 30 Brown Swiss (Bos taurus ) bulls. Semen was collected by using an artificial vagina and by electroejaculation; the ejaculates were individually evaluated. Semen was diluted for freezing in skimmed milk and stored in 0.5-ml French straws, at a concentration of 30 x 10(6) spermatozoa. Data were evaluated by analysis of variance using a factorial model which included collection method, breed effect, and interaction between method and breed, with each bull as a block. Higher volume and pH of the semen was obtained following electroejaculation. Conversely, higher concentration prior to freezing and better progressive motility after thawing was observed in semen collected with an artificial vagina. No differences in motility were obtained in fresh semen between methods. Better post-thaw recovery occurred when the semen was collected by an artificial vagina, independently of the breed type.     

Relationships between Brucella ovis semen culture and various semen and serology parameters

Semen and blood samples from 154 rams from two Montana range flocks (Flock A, vaccinated for Brucella ovis ; Flock B, nonvaccinated) were evaluated to determine the relationship between Brucella ovis (B. ovis ) semen culture results and various semen and blood parameters. All rams utilized in this study exhibited no palpable ram epididymitis lesions. Thirteen and 25.6% of the rams tested in Flocks A and B, respectively, had positive B. ovis semen cultures. Only age of ram and ram condition scores differed (P<0.05) between flocks. No flock by semen culture interactions were detected (P>0.05) for any of the parameters evaluated. Age of ram, ram condition score, and spermatozoa rate from forward movement were unrelated (P>0.05) to B. ovis culture results. Rams with positive B. ovis semen cultures had lower sperm motility (P<0.05), higher percentage of abnormal spermatozoa cells (P<0.05), higher percentage of spermatozoa head abnormalities (P<0.01), lower percentage of live-normal cells (P<0.05), higher incidence of white blood cells in semen (P<0.01) and higher complement fixation (CF) titers (P<0.01).     

Quality and freezing qualities of first and second ejaculates collected from endangered Gulf Coast Native rams

The Gulf Coast Native sheep, or Louisiana Native sheep, is an endangered previously feral domestic sheep population of European origin that has been under natural selection pressure for reproductive survival in their transplanted range while roaming in the southern Gulf Coast Region of the United States. This sheep population has an increased natural resistance to internal parasites, breeds year-around and has a greater percentage of live lambs as compared with other breeds of sheep raised in similar environments. To preserve the genetic diversity of this important feral sheep population, semen was collected by electro-ejaculation and subjected to cryopreservation for subsequent storage in a genome resource bank. Unrelated rams (n=5) were collected 3 days-a-week, allowing at least 2 days of rest between collections. Two ejaculates were obtained from each ram per collection day, with the second collection conducted 10min after the first ejaculation. Semen was processed using the standard Salamon cryopreservation procedure in a Tris-yolk-glycerol extender, frozen in 0.5ml plastic straws using liquid nitrogen (LN(2)) vapor and stored in LN(2). Each ejaculate was evaluated for volume, sperm concentration/ml (x10(9)/ml), number of spermatozoa/ejaculate (x10(9)), sperm progressive motility (%) for pre-cooled semen, cooled semen and semen after thawing. For the five rams, each semen variable for the first ejaculate was compared with that of the second ejaculate collected 10min later. The mean semen volume, sperm concentration and number of spermatozoa per ejaculate obtained from the first ejaculate were significantly greater (P< or =0.01) than those of the second ejaculate (comparisons being 1.62 and 1.06; 3.2 and 1.5; 5.4 and 1.8, respectively). Overall, the mean motility of pre-cooled (22 degrees Celsius), cooled (5 degrees Celsius) and frozen (-196 degrees Celsius) post-thawed spermatozoa was less (P< or =0.01) in the first ejaculate (71.5, 64.8 and 34.1%, respectively) compared with that of the second ejaculate (75, 72.4 and 44.1%, respectively). Conversely, no differences were detected in loss in the percent progressive motility of sperm from cooled sperm to post-thaw sperm from the first and second ejaculates. In summary, our findings suggest sperm collected during the second ejaculate 10min after the first ejaculate of rams survives thawing with a greater rate of progressive motility than that of the first ejaculate. The ability to collect two consecutive ejaculates in a short period by electro-ejaculation could be valuable for gamete resource banking and preserving genetic diversity of the Gulf Coast Native sheep.     

Semen characteristics of the guinea fowl Numida meleagris meleagris

Eighteen adult exotic Golden Sovereign guinea fowl (Numida meleagris meleagris) males identified by leg bands and housed individually in cages were ejaculated two times a week at 4- and 3-d intervals (Mondays and Thursdays) for 6 wk. Semen was obtained manually by gently massaging the dorso-lateral lumbo-sacral region. Semen collection and evaluation were done between 1400 and 1800 h each day of test. Mean semen volume was 0.032 +/- 0.001 ml, sperm motility was 37.1 +/- 0.1% and sperm concentration/ml was 2.62 +/- 0.01 x 10(9). Percentage live sperm averaged 91.6 +/- 0.1, while the mean percentage morphologically normal spermatozoa was 76.9 +/- 0.5. Primary and secondary sperm abnormalities were 11.9 +/- 0.2% and 11.3 +/- 0.2%, respectively. Birds differed significantly in all ejaculate characteristics studied except percent secondary abnormalities, indicating that considerable variation exists for improvement of these semen traits. There were significant bird x collection interval (P<0.05) and bird x week (P<0.01) interactions for sperm concentration/ml and bird x week interaction for sperm motility (P<0.05). The results generally are within acceptable levels and show that the semen is suitable for use in artificial insemination.     

Semen quality in dogs and the influence of a short-interval second ejaculation

Semen quality was examined in each of 65 known fertile dogs. Values were found to be similar to those previously published, although an apparent breed influence was demonstrated, with German shepherd dogs producing ejaculates of larger volume and greater total spermatozoal output than other breeds. A second ejaculate was collected from each dog with a mean interval of 63 min. The second samples had significantly lower values for the volume of the second fraction, the spermatozoal concentration and the total spermatozoal output. There were no differences for the percentage motility or the percentage of morphologically normal live spermatozoa. While there was no increase in semen quality of the second ejaculate, the technique may be useful since it results in the collection of approximately 70% more spermatozoa than a single ejaculate. These spermatozoa also had normal motility and morphology and could, therefore, be used for insemination or cryopreservation.     

Post-thaw functional status of boar spermatozoa cryopreserved using three controlled rate freezers: a comparison

This study compared variation in the quality of cryopreserved boar spermatozoa and the control and accuracy of cooling rates between three semen freezers (CryoLogic Freeze Control CL3000, Planer Products Kryo Save Compact KS1.7/Kryo 10 Control module and a controlled rate 'Watson' freezing machine developed within our laboratory). Five ejaculates were collected from each of 15 boars (five boars from each of three breeds). Semen was diluted into a commercial freezing buffer (700 mOsm/kg, 3% v/v glycerol) and placed into 0.5 ml straws. Three straws per treatment, from each ejaculate were cooled to -5 degrees C at 6 degrees C/min, held at -5 degrees C for 30s while ice crystal formation was induced, then further cooled from -5 to 80 degrees C at either 40 degrees C/min (Kryo Save Compact KS1.7 and Watson) or 6 degrees C/min (Freeze Control CL3000). Precise measurements of temperature fluctuations during the programmed cooling curves were made by inserting thermocouples into the semen filled straws. Semen was assessed for %motile cells, motility characteristics using computer-assisted semen analysis (CASA), plasma membrane integrity (%SYBR-14 positive stained spermatozoa) and acrosome integrity (%FITC-PNA positive stained spermatozoa). Spermatozoa cryopreserved using the Freeze Control CL3000 system (maximum rate of 6 degrees C/min) exhibited reduced post-thaw viability (14.2+/-2.8% mean plasma membrane intact spermatozoa) when compared to both the KS1.7 and Watson freezers (optimal rate of 40 degrees C/min) (18.4+/-3.2 and 25.7+/-3.7% mean plasma membrane intact spermatozoa, respectively). Differences in motility characteristics were observed between spermatozoa cryopreserved at 40 degrees C/min with the Watson apparatus preserving a larger proportion of sperm with progressive motility. Cooling curves in the CL3000 and KS1.7 were interrupted by a pronounced increase in temperature at -5 degrees C that corresponded with the latent heat of fusion released with ice crystal formation. This temperature change was significantly reduced in the cooling curves produced by the Watson freezer. These findings suggest that preserving spermatozoa using the Watson freezer improved post-thaw semen quality, with regard to sperm motility characteristics. Furthermore, that post-thaw semen viability was enhanced by minimising temperature fluctuations resulting from the release of the latent heat of fusion at ice crystal formation.     

Seminal traits, suitability for semen preservation and fertility in the native Portuguese horse breeds Puro Sangue Lusitano and Sorraia: Implications for stallion classification and assisted reproduction

The Puro Sangue Lusitano (PSL) is the major national breed of horse in Portugal, but no studies exist on its seminal characteristics, or on the possibility of conserving semen for future use. The aim of this study was to evaluate semen parameters, fertility and the aptness to semen preservation in Lusitano Stallions. In order to compare characteristics defined by a single or by multiple semen collections per stallion 152 ejaculates obtained from 152 Lusitano stallions presented at an annual breeding soundness examination as well as data related to 371 ejaculates obtained from 9 PSL were analyzed. These latter samples were also evaluated in terms of their possible use in assisted reproduction and were compared with 113 ejaculates obtained from 4 Sorraia horses, a rare and endangered Portuguese breed. The percentage of motile spermatozoa (PMS) was assessed after collection (AC), after semen dilution (AD) and at 24h of cool-storage. Mean values obtained for sperm motility and morphology and semen pH observed after semen collection differ significantly (P<0.05) between single collection/multiple stallions and multiple collections/limited stallions, and no age related effects were detected. Overall, Lusitano semen quality was comparable to that of related breeds, while Sorraia stallions had very poor semen quality. The response to cool-storage of diluted semen samples differed among stallions and breeds, and the best results for progressive motile sperm cells at 24h were in a range of 35-53% for PSL stallions and were lower for Sorraia stallions. Fertility rates obtained with artificial insemination (AI) averaged at 85% for PSL. With the exception of PMS AC, sperm vitality and semen pH no other seminal trait seemed to influence fertility rates in the Lusitano breed.     

The relationship between rabbit semen characteristics and reproductive performance after artificial insemination

The relationships between several rabbit buck semen traits, concerning either the ejaculate or the dose inseminated (volume, mass motility, pH, percentage of motile sperm (PMS), concentration, number of total and motile sperm per ejaculate and per insemination dose) and the reproductive performance of does was investigated in 839 inseminations involving 54 bucks and 111 does. Four genetic types were involved: INRA1601 strain (A), INRA2066 strain (B) and the two reciprocal crossbreds (AB and BA). The mating design was A x A, B x B, (AB or BA) x (AB or BA). Semen was diluted (1:9) and a constant volume of 0.5 ml was inseminated 2-4h after collection. Therefore, the total number of spermatozoa per dose was variable and proportional to the initial concentration. Mass motility significantly influenced the kindling rate. Taken separately, volume, PMS and concentration did not influence the kindling rate but their product, the number of motile sperm per ejaculate, did. Litter size (total born) was significantly influenced by concentration and all variables depending on it, particularly the number of total and motile sperm per dose. However, reproductive performances were predominantly influenced by the physiological status of the does at insemination (lactation stage and receptivity).