Seminal Characteristics in Spanish Purebred Stallions: A Retrospective Study

In this work, we studied retrospectively the influence of several factors on semen quality of the Spanish Purebred (SPB) stallion. Among these factors, we considered stallion age, season, and interval between two consecutive collections. The semen was collected from 11 SPB stallions (634 ejaculates). Semen quality was evaluated based on volume, concentration, and motility of the ejaculates. Results showed that the semen collected in autumn and winter was more concentrated in comparison with that collected in spring and summer, when the volume of ejaculate was the highest. Those ejaculates collected from younger stallions were characterized by having the smallest volume and the highest sperm concentration. When incorporating an SPB stallion into a program of artificial insemination, it is important to take into account all of these factors to achieve the maximum reproductive potential of the animal.     

Effects of seasons on some semen parameters and bacterial contamination of Awassi ram semen

The objectives of the present study were to determine the effects of season on some semen parameters and bacterial contamination of Awassi ram semen. Semen samples from six mature Awassi rams were used in this study. Semen collection was performed with artificial vagina every week, from September 2009 to October 2010. Volume, sperm concentration, mass motility, individual motility, percentage live sperm and sperm abnormalities were evaluated. Moreover, determination of viable bacterial count of the rams was also recorded weekly. Higher (p < 0.05) semen volume in the hot summer and spring months was observed of August (1.55 ± 0.08 ml) and March (1.27 ± 0.15 ml). Sperm concentration was highest (p < 0.05) in the breeding season (late summer to early autumn) of September (4.21 ± 0.86 × 10(9) sperm/ml). Sperm individual motility and percent of live sperm observed in August (summer) and May (end of spring) when the environmental temperature started to increase were recorded highest values and differed significantly (p < 0.05) from December and January (winter). The highest value of the mean sperm acrosomal defects (13.33 ± 0.63%) was recorded in December. The highest value of the mean viable bacterial count (138.3 ± 21.6) was recorded in July (summer). A significant decrease (p < 0.01) in the mean viable bacterial count was observed from the middle of winter towards the end of spring. The lowest bacterial count was noted in January (60.5 ± 2.98). It could be concluded from the results of the present study that there is an effect of season on ram semen quality, and summer high temperature in northern Iraq has no effect on Awassi ram semen. There is a significant effect of season on bacterial count on Awassi ram semen.     

The Effect of Season on Semen Characteristics and Freezability in Bos indicus and Bos taurus Bulls in the Southeastern Region of Brazil

The objectives of this study were to evaluate the effects of season in southeast of Brazil comparing genotypes on semen characteristics, freezability and peripheral plasma concentrations of testosterone. Ejaculates of five Bos indicus bulls and six Bos taurus bulls were evaluated over a period of 27 months, which was divided into winter (July, August, September), spring (October, November, December), summer (January, February, March) and autumn (April, May, June). Semen was evaluated according to standard procedures for ejaculate volume, sperm concentration, gross-motility, progressive motility and sperm morphology. After preparing and freezing the ejaculates according to commercial procedures, the straws were stored in liquid N₂ until post-thaw evaluation. Ejaculate volume, sperm concentration, gross-motility, progressive sperm motility, vigor and morphological sperm defects were significantly influenced by season and genotype (p < 0.05). Heat tolerance was better in B. indicus bulls than in B. taurus bulls characterized by lower values of sperm abnormalities throughout the observation period. The highest values were recorded for abnormal heads followed by cytoplasmatic droplets in B. taurus bulls. The proportion of ejaculates which were eliminated before freezing for reasons of bad quality was lower in the B. indicus bulls. Temporal changes in peripheral plasma testosterone concentrations were higher in B. indicus bulls than in B. taurus bulls not revealing seasonal influences. The results of this study show clear genotype differences regarding semen quality. Freezability of B. taurus semen varies considerably throughout the year, leading to a high proportion of eliminated ejaculates. Collecting semen from B. taurus bulls during the summer in an artificial insemination centre may not be profitable.     

Influence of Seasons, Extenders, Slow and Rapid Freezing on Seminal Characters in Korean Native Bucks

The success of AI technology is based on both semen quality and freezing process. In order to establish the semen freezing techniques in Korean native bucks, factors affecting the success were evaluated in the present study. Semen collected by electro-ejaculation from bucks during four distinct seasons was evaluated for semen volume and pH, sperm motility and survivability. The semen volume, concentration and total cell were higher in spring, summer and less in winter. Yet, there were no seasonal differences in the proportional data of live sperm, motility score and pH of semen among seasons. The percentage of live sperm after thawing was found to increase with increased concentration of lactose in Tris-Egg yolk-glycerol (TY-G), being highest in TY-G supplemented with 180 mm lactose (TYL180-G), but did not differ between TY-G and TYL120-G. Sperm motility was enhanced by employing 2.0 h equilibration time with rapid freezing method. In conclusion, semen could be frozen with high success rates for further use of AI in breeding techniques and to preserve the Korean native bucks.     

Melatonin can improve viability and functional integrity of cooled and frozen/thawed rabbit spermatozoa

Melatonin is known to protect sperm against freezing-inflicted damage in different domestic species. The aim of the study was to evaluate the effect of supplementation of semen extender with melatonin on the quality and DNA integrity of cooled and frozen/thawed rabbit spermatozoa. We also investigated whether the addition of melatonin to the semen extender could improve the fertility of rabbit does artificially inseminated with frozen/thawed semen. Semen samples collected from eight rabbit bucks were pooled and then diluted in INRA-82 supplemented either with (0.5, 1.0 or 1.5 mM) or without (0.0 mM) melatonin. Diluted semen was cooled at 5°C for 24 hr. For cryopreservation and based on the first experiment's best result, semen samples were diluted in INRA-82 in the presence or absence of 1.0 mM melatonin and then frozen in 0.25 ml straws. Following cooling or thawing, sperm quality and DNA integrity were evaluated. Furthermore, the fertility of frozen/thawed semen was investigated after artificial insemination. Supplementation of semen extender with 1.0 mM melatonin improved (p < .05) motility, viability, membrane and acrosome integrities in cooled semen compared with other groups. Sperm quality and DNA integrity were higher (p < .05) in frozen/thawed semen diluted in 1.0 mM melatonin-supplemented extender than in the control group. Conception and birth rates were higher in does inseminated with 1.0 mM melatonin treated semen compared with the controls. In conclusion, supplementation of semen extender with 1.0 mM melatonin improved the quality of cooled and frozen/thawed rabbit spermatozoa. Melatonin can preserve DNA integrity and enhance the fertility of frozen/thawed rabbit spermatozoa.     

Effect of semen and donor factors on multiple ovulation and embryo transfer (MOET) in sheep

Multiple ovulation and embryo transfer (MOET) is an important tool in the sheep industry for increasing numbers of genetically superior individuals. The objective of this study was to evaluate the effect of semen source (frozen or fresh), the number of embryo collection procedures for each donor (NECP), the season in which embryo transfer and collection was performed, and the age and breed of the donor, on the number of recovered embryos and pregnancy rates after embryo transfer. The Alamos Genetics’ flushing station database was used. This consisted of 140 embryo collection procedures, from 53 Dorper and White Dorper sheep donors, aged between one and eight years, totalling 1,200 collected embryos. Neither the number of retrieved embryos nor the pregnancy rate was affected by the semen preservation method (fresh or frozen), NECP or the age and breed of donor. The season did not affect the number of collected embryos but had a significant effect (p < 0.05) on the recipient pregnancy rate, with higher pregnancy rates reported in the winter (65.57% ± 25.33%) compared with spring (37.11% ± 33.27%), summer (29.95% ± 28.33%) or autumn (35.03% ± 31.66%). There is an estimated increase of 98.4% and 71.5% of embryos recovered in the spring and summer seasons, respectively, when winter is used as reference. The survival of embryos is significantly higher when implanted during the breeding season, more specifically in winter. Embryo collection can be carried out throughout the year in sheep, but there may be a marginal advantage in the use of superovulation and fresh embryo transfer programmes in the autumn and winter.     

Effect of Collection Rhythms and Season on Rabbit Semen Production

The effects of collection regimen and time of year on rabbit semen production were determined in this study. A total of 14 crossbreed Hyla bucks were used in winter and summer. In each season, rabbits were assigned to two groups. In group 1, (n = 7) rabbits were subjected to an extensive collection regimen (two ejaculates per male, once daily/week) and in group 2, (n = 7) a semi‐intensive semen collection regimen was performed (two ejaculates per male, twice weekly). The traits recorded for each sample were libido, volume, pH, motility, sperm concentration, percentage of alive spermatozoa and sperm abnormalities. The results obtained in this study indicate that when increasing collection frequency, the rate of useful collections decreased (from 0.81 ± 0.017 to 0.69 ± 0.016; p < 0.01). The rate of useful collection also decreased in the transition from winter to summer (from 0.79 ± 0.018 to 0.70 ± 0.017; p < 0.01). Among the ejaculate characteristics studied, only volume/ejaculate (from 0.64 ± 0.015 to 0.53 ± 0.017; p < 0.01) and spermatozoa/ml (from 406 ± 15 to 359 ± 13 million; p < 0.01) appeared negatively affected by collection. In winter fewer volume/ejaculates were produced (0.55 ± 0.015 vs 0.60 ± 0.016 ml; p < 0.01) and fewer spermatozoa/ml (360 ± 14 vs 394 ± 16 million; p < 0.01) than in summer. The doses produced per ejaculate decreased as collection frequency increased, but the number of doses produced per week was higher in the semi‐intensive than the extensive rhythm (26.5 ± 2.1 vs 20.9 ± 1.5; p < 0.01). The results suggest that a semi‐intensive rhythm may be viewed favourably.     

Combined cryopreservation of canine ejaculates collected at a one-hour interval increases semen doses for artificial insemination without negative effects on post-thaw sperm characteristics

A limiting factor in canine artificial insemination (AI) is the low number of insemination doses obtained per ejaculate. In this study, semen was collected from dogs (n = 28) either once and frozen directly after collection or the same dogs were submitted to a dual semen collection with a 1-hr interval and the two ejaculates were combined for cryopreservation. We hypothesized that combining two ejaculates increases semen doses per cryopreservation process without negative effects on semen characteristics. Total sperm count was lower in semen from a single semen collection in comparison with the combination of the first and second ejaculate of a dual semen collection (p < .001). The percentage of motile and membrane-intact spermatozoa determined by computer-assisted sperm analysis (CASA) in raw semen did not differ between single and combined dual ejaculates and was reduced (p < .001) by cryopreservation to the same extent in single (motility 73.7 ± 1.8%, membrane integrity 65.6 ± 2.2%) and combined dual ejaculates (motility 72.7 ± 2.3%, membrane integrity 64.6 ± 2.5%). The percentage of spermatozoa with morphological defects increased after cryopreservation (p < .001) but was similar in single and combined dual ejaculates. The CASA sperm velocity parameters decreased with cryopreservation (p < .001) but did not differ between single and combined dual ejaculates. The number of insemination doses increased from 2.7 ± 0.4 for single to 4.7 ± 0.8 for combined dual ejaculates (p < .01), based on 100 million motile spermatozoa per frozen-thawed semen dose. In conclusion, combining two ejaculates collected at short interval for one cryopreservation process increases the number of AI doses without compromising semen quality.     

Multivariate models for estimating jackass semen production and quality

The purpose of this study was to analyse the effects of season, age, gonad and accessory sex glands on semen characteristics of jackass and to construct multivariate regression models to predict semen quality. In autumn, spring and summer, semen characteristics of 30 sexually mature donkeys (1,014 ejaculations) were analysed to investigate the effect of seasons on semen quality, and gonad and accessory sex gland parameters of 12 jackasses were measured immediately after ejaculation by ultrasonography to investigate the effect of seasons on reproductive organ size. Semen (598 ejaculates), gonad and accessory sex gland parameters of 40 jackasses aged between 3 and 7 years were analysed in autumn to investigate the effects of age and reproductive organ size on semen quality and to construct multivariate models. To verify the accuracy of the models, semen (476 ejaculates), gonad and accessory sex gland parameters of 20 jackasses were measured from March to June. Results revealed that semen, gonad and accessory sex gland parameters were not affected by season and age. Progressive motility (PM) was positively correlated with long axis of the spermatic cord (LASC) and negatively correlated with percentages of sperm abnormality (PSA). Total sperm count (TSC) was positively correlated with testis circumferences (TC) and cross-sectional area of cauda epididymis (CSACE). TC, CSACE, LASC and PSA were included into multivariate models to predict PM, TSC and functional sperm count (FSC) in 20 jackasses (PM = 72.332 + 0.428 LASC − 0.441 PSA; TSC = −169.929 + 8.728 TC + 0.253 CSACE; FSC = −206.645 + 8.788 TC + 0.258 CSACE). The predicted and observed values corresponded well. In conclusion, the tested models can be used for predicting semen quality of donkey.     

Effect of scrotal bifurcation on breeding soundness traits in Beetal bucks during summer season

Relationship of scrotal bifurcation or splitness with breeding soundness traits was investigated in 15 Beetal bucks (17–20 months of age and 48.72?±?1.57 kg mean BW). Breeding soundness traits of conjoined/unsplit (n?=?6) scrotal bucks was compared with split (n?=?9) scrotal bucks having lengthwise >?1 in. bifurcation. Two consecutive semen ejaculations per buck were collected at monthly interval during summer season (April to June 2018) using intact buck as teaser and sexual behavior was simultaneously recorded. Scrotal morphometry parameters, i.e., scrotal circumference, scrotal volume, scrotal skin thickness, testis length, width, and thickness were also recorded. Bucks with split scrotum had relatively more scrotal dimensions (scrotal circumference (P?<?0.01), scrotal volume (P?<?0.01), testis length (P?<?0.01)) than conjoined scrotal bucks. Among various semen attributes, semen volume of first ejaculate (P?<?0.01), second ejaculate (P?<?0.05), mean semen volume (P?<?0.01), total sperm count of first ejaculate, and mean sperm count of both ejaculates were more (P?<?0.01) in split scrotal bucks. However, scrotal bifurcation had no influence on sexual behavior of bucks. It is concluded that Beetal bucks with split scrotum had relatively better breeding efficiency traits than conjoined scrotal bucks.

     

Monitoring of libido and semen quality parameters in melatonin‐treated French alpine bucks during the non‐breeding season

The aim of this study was to investigate the effects of exogenous melatonin on libido and semen quality parameters in bucks during the non‐breeding season. Twelve bucks of the French alpine breed from 1.5 to 4 years of age were assigned into melatonin (MG) and control (CG) groups, with 6 bucks in each group. The experimental period was 3 months (March–May), divided into six periods of 15 days each. The bucks in the MG group received four melatonin implants at the end of March. Two semen samples were taken from the bucks by artificial vagina once per week and their libido estimated. Volume and spermatozoa concentration, their mass motility and motility, proportion of live and total abnormal and forms with abnormal head and tail were determined in the obtained ejaculate samples. The total number of spermatozoa and functional spermatozoa fraction in the ejaculate was also calculated. The MG bucks had significantly higher mass motility and motility of spermatozoa in the first half of April, and a higher proportion of live spermatozoa in the first and second half of April (p < .05). Differences in libido intensity were not significant. The results indicated that the application of melatonin significantly improved the qualitative parameters of semen in bucks, as seen in increased mass motility, motility of spermatozoa and proportion of live spermatozoa shortly following melatonin insertion. Therefore, the results of the current study are novel regarding the use of melatonin treatment during the non‐breeding season to improve the qualitative parameters of ejaculates in bucks.     

Motility and Fertility Evaluation of Thawed Frozen Stallion Semen After 24 Hours of Cooled Storage

Breeding mares with cryopreserved semen requires specialized equipment for storage and thawing and more intensive mare management. The objectives of this study were (1) evaluate the longevity of frozen stallion semen once it had been thawed, extended, and maintained at 5°C for 48 hours in a passive cooling container, and (2) determine fertility potential of frozen semen that had been thawed, extended, and used to inseminate mares after 24 hours of cooled storage. Eight ejaculates were collected and aliquots were cooled in either INRA96 and CryoMax LE minus cryoprotectant at a concentration of 50 million total sperm/mL. The remainder of the ejaculate was frozen in CryoMax LE extender at a concentration of 200 million total sperm/mL. Semen was thawed using 1 of 3 thawing protocols, and diluted to a concentration of 50 million total sperm/mL in either INRA96 or CryoMax LE minus cryoprotectant and cooled to 5°C. Sperm motility was evaluated at 24 and 48 hours. Eight mares were inseminated over two estrous cycles using frozen semen that had been thawed, extended in INRA96, and cooled for 24 hours. There was no difference in progressive motility at 24 or 48 hours of cooled-storage post-thaw between the 3 thawing protocols. An overall per cycle pregnancy rate of 56% (9/16 cycles) was achieved using frozen-thawed semen that had been extended and cooled for 24 hours. In summary, frozen stallion sperm was thawed, extended, and cooled to 5°C for 24 hours and still maintained adequate (>30%) sperm motility and fertility.     

Estimation of Genetic Parameters and Analysis of Season Effects and Age Effects for Semen Traits in Landrace Boars

The purpose of this study was to estimate the genetic parameters of semen traits of Landrace boars in South China and to analyze the influence of the age and season of semen collection on semen traits, so as to provide theoretical basis for formulating reasonable boar breeding program. The Asreml-R was used to analyze the 107 221 semen data of 1 605 Landrace boars from 2 AI stations in Southern China, the single-trait repeatability animal model was used to estimate the variance components, heritability and repeatability of each semen traits, two-trait repeatability animal model was used to estimate genetic correlation and phenotypic correlation of semen volume, semen density, sperm motility and percentage of abnormal sperm. The general linear model of R language program was used to analyze the influence of age and season of semen collection on semen traits. The results showed that the semen volume and percentage of abnormal sperm had medium heritability (0.23 and 0.38), and the variation coefficient of percentage of abnormal sperm was 85.42%, while the other traits had low heritability (0.07-0.19). Semen volume-semen density and sperm motility-percentage of abnormal sperm had extremely significantly negative genetic correlation(-0.77 and -0.90, respectively). Semen density-sperm motility showed extremely significantly positive genetic correlation (0.50). The influence of the age of semen collection on semen traits was significant(P<0.05). After boars reached sexual maturity, semen volume showed a significant increase, semen density and percentage of abnormal sperm showed a general decrease, and total sperm number and functional sperm number were significantly higher in the 13-18-month of age group than those in the other groups (P<0.05). The semen density was the highest in spring, and the total sperm number and functional sperm number were significantly higher in autumn and winter than those in spring and summer (P<0.05). In conclusion, the semen volume and percentage of abnormal sperm of Landrace boars can be selected as candidate traits. In the boar production management, boar whose semen production after 36 months of age decline, will be considered to update. Boar semen quality was significantly higher in autumn and winter than those in spring and summer. It is suggested that cooling work should be done in advance in Southern China in summer.     

Effects of supplemental conjugated linoleic acids (CLA) on fresh and post‐thaw sperm quality of Holstein bulls

This study was designed to investigate the effects of feeding‐protected conjugated linoleic acid (CLA) on the semen production and sperm freezability in Holstein bulls. Twelve bulls were randomly assigned to two groups (n = 6 per group). Bulls received the normal diet (control group) or the normal diet top‐dressed with 50 g of CLA (treated group) for 10 weeks. The control group received 40 g/day calcium soap of fatty acid. Fresh and post‐thaw semen quality was assessed on ejaculates collected at the 0, 4, 6, 8 and 10 week of supplementation. Semen evaluations including sperm concentration, motion characteristics (subjective and computer‐assisted), viability (Eosin–Nigrosin), membrane integrity (hypo‐osmotic swelling test) and abnormality were conducted. Semen volume, sperm concentration and total sperm output were not affected by dietary treatment (p > .05). The proportion of spermatozoa with abnormal morphology in fresh semen significantly increased (p < .05) in the CLA‐fed group compared to control group. Also, in CLA‐fed group, the proportion of post‐thaw spermatozoa with abnormal morphology at week 10 of trial was significantly higher in CLA than control group (p < .05). Progressive motility tended to be increased in the CLA‐fed group, although dietary supplementation did not affect other CASA parameters or viability in fresh and frozen‐thawed sperm. In this study, CLA supplementation had little positive effect on fresh or post‐thaw sperm quality of Holstein bulls.     

长白公猪精液性状遗传参数评估及采精月龄和季节因素分析

旨在估计华南地区长白种公猪精液性状遗传参数以及分析公猪采精月龄和季节对精液性状的影响,为合理制定种公猪育种方案提供理论依据。本研究采用Asreml-R对华南地区两个公猪站1 605头长白公猪107 221条精液数据进行统计分析,利用单性状重复力动物模型估计公猪精液各性状的方差组分、遗传力和重复力,利用两性状重复力动物模型对精液体积、精液密度、精子活力和精子畸形率等性状进行遗传相关和表型相关估计;利用R语言程序中的一般线性模型分析采精月龄和季节对精液性状的影响。结果表明,精液体积和精子畸形率属于中等遗传力（0.23和0.38）,其中精子畸形率的变异系数较大,为85.42%,其余性状都为低遗传力（0.07~0.19）;精液体积与精液密度以及精子活力与精子畸形率为极显著遗传负相关（-0.77和-0.90）;精液密度与精子活力呈极显著的遗传正相关（0.50）。采精月龄对精液性状影响显著（P<0.05）,在公猪达到性成熟后,精液体积呈显著上升趋势,精液密度和精子畸形率总体呈下降趋势,精子总数和有效精子数在13~18月龄组显著高于其他组（P<0.05）。在春季精液密度最高,精子总数和有效精子数在秋、冬季显著优于春、夏季（P<0.05）。综上,长白公猪精液体积和精子畸形率具有较大的选育潜力,可作为候选性状进行选择。在公猪生产管理方面,公猪36月龄后精液产量下降,考虑更新淘汰。公猪精液品质在秋、冬季显著优于春、夏季,在中国南方夏季应提前做好降温工作。     

Testosterone serum profile,semen characteristics and testicular biometry of Mangalarga Marchador stallions in a tropical environment

This study was conducted to characterize the daily profile of testosterone secretion and its mean concentrations in the four seasons as well as to evaluate the semen characteristics and testicular biometry of Mangalarga Marchador stallions throughout the year in a tropical region. Three stallions were submitted to semen collections and evaluation of testicular biometry every 14 days along a year. Blood samples were collected once at the middle of each season, in a 20‐min interval during 24 hr in order to evaluate the testosterone secretion profiles among seasons. Testosterone concentrations along the day were higher at the beginning of the afternoon (from 12:00 to 15:00 hr), but a circadian secretion was not clearly observed. Mean testosterone concentrations did not differ among seasons (p > .05), but a pattern of secretion along the day showed variations with higher concentrations in the afternoon during the winter. Ejaculate volume was higher during summer; however, sperm motility decreased in summer and spring. Total sperm in ejaculate, sperm morphology and testicular biometry kept constant along the year showing no differences among the seasons. The results demonstrated that in a tropical region, reproductive aspects of stallions did not show a clearly defined seasonal variation, and months of autumn and winter were not unsuitable for reproduction of the males.     

Sperm Production and Sperm Morphology of Swedish Warmblood Stallions

The present study investigated daily sperm output and sperm morphology of fresh semen in eight Swedish Warmblood stallions aged 5–8 years. They were used for artificial insemination, and their fertility during the breeding season of semen collection exceeded 60% per cycle. One ejaculate of semen was collected daily for 10 consecutive days from each stallion. The gel-free volume was measured, and the sperm concentration was assessed with a Bürker chamber. The volume of gel-free fraction was multiplied by the sperm concentration to give the total number of spermatozoa (TSN). Sperm morphology was examined in ejaculates collected on days 2, 5 and 10. An aliquot from each ejaculate was fixed in 1 ml formol–saline immediately after collection and examined under a phase-contrast microscope (magnification 1000×) to assess morphological abnormalities. Furthermore smears were prepared and stained according to Williams (carbolfuchsin–eosin) for a more detailed examination of the sperm heads under a light microscope (magnification 1000×). Analysis of variance was applied to data. Total spermatozoa number decreased progressively during the first 8 days of collection, and daily sperm output (DSO) was calculated as mean TSN of collections on days 8–10, being 6.4 × 10⁹ spermatozoa. The overall percentages of morphologically normal spermatozoa in ejaculates collected on days 2, 5 and 10 were above 70%, being significantly lower in ejaculate 2 (68.6%) compared with ejaculates 5 and 10 (72.9% respectively 75.3%).     

Influence of the Preservation Temperature (37, 20, 4, −196°C) and the Mixing of Semen over Sperm Quality of Majorera Bucks

This study assessed the effect of different semen storage temperatures and the influence of semen pooling in semen viability. In experiment 1, semen samples (n = 30) of five Majorera bucks were individually processed [Individual semen (IS)] and after the first dilution (Tris‐yolk extender), semen‐diluted aliquots from each male were pooled semen (PS). Thereafter, semen samples (IS and PS) were preserved as fresh semen (37 and 20°C), chilled semen (4°C) and frozen semen. Sperm motility and the percentage of abnormal sperm cells and intact membrane acrosomes were defined. Semen preservation at 20 and 4°C did not modify the quality of spermatozoa for the first 24 h, but the conservation at 37°C caused a dramatic fall in the semen motility from 12 h onwards. Furthermore, the longevity of frozen‐thawed semen was limited to 4–6 h. No differences were observed in semen parameters when PS was compared with semen from individual males in any of the preservation protocols assessed. In experiment 2, 120 goats were distributed in four experimental groups: in group fresh individual semen (FIS, n = 30) and group frozen‐thawed individual semen (FTIS, n = 30), does were transcervically inseminated with fresh semen and frozen‐thawed semen from each individual male, respectively, and in group fresh pooled semen (FPS, n = 30) and group frozen‐thawed pooled semen (FTPS, n = 30), goats were transcervically inseminated with FPS and FTPS, respectively. The kidding rate was very close in the FIS and FPS groups (70.0% and 73.7%, respectively), and no significant differences were observed in the fertility rate between FTIS and FTPS. The results of this study confirmed that semen samples may be preserved satisfactorily for 24 h both at 20 and 4°C. In addition, the mixture of semen of different bucks did not significantly modify the semen parameters when compared with semen from individual males.     

Seminal Plasma Characteristics and Expression of ATP‐binding Cassette Transporter A1 (ABCA1) in Canine Spermatozoa from Ejaculates with Good and Bad Freezability

The composition of seminal plasma and the localization of the ATP‐binding cassette transporter A1 (ABCA1) in spermatozoa from good and bad freezers were compared to frozen–thawed spermatozoa from the same dog. Ejaculates were obtained from 31 stud dogs, and the sperm‐rich fraction (SRF) was kept for analysis. One aliquot was used for the analysis of concentration, progressive motility (P; CASA), viability (V; CASA) and leucocyte count, and the analysis was performed by flow cytometry (FITC‐PNA/PI), SCSA and HOST. In seminal plasma, concentration of albumin, cholesterol, calcium, inorganic phosphate, sodium, potassium, zinc and copper was measured. Semen smears were prepared and evaluated for the expression of ABCA1. The remainder of each ejaculate was frozen. After thawing, the quality assessment was repeated and further smears were prepared. According to post‐thaw semen quality, dogs were assigned to good freezers (n = 20) or bad freezers (n = 11), the latter were defined as < 50% progressive motility and/or > 40% morphologically abnormal sperm and/or < 50% viability. Bad freezers were older than good freezers (5.3 vs 3.4 years, p < 0.05). In bad freezers, the percentage of sperm with ABCA1 signal in the acrosome was lower (26.3% vs 35.7%, p < 0.01) and the percentage of sperm with complete loss of ABCA1 signal higher (46.7% vs 30%, p < 0.01); the percentage of dead spermatozoa was higher (36.1% vs 25.5%, p < 0.05), and the concentration of cholesterol and sodium in seminal plasma was lower than in good freezers (p < 0.05). We conclude that in thawed bad freezer sperm, an increase in acrosome damages coincided with an increased loss of cholesterol transporters and cell death, and a lower cholesterol concentration in seminal plasma. Follow‐up studies revealed whether a relation exists between these findings.     

Influence of constant long days on ejaculate parameters of rabbits reared under natural environment conditions of Mediterranean area

Since rabbit bucks are usually housed under constant long daylight in artificial insemination (AI) centers, the main purpose of this study was to investigate whether constant long day influenced ejaculate parameters of rabbits housed in AI centers in the Spanish Mediterranean area. The study was carried out in Murcia, Spain (37° N). Twenty commercial hybrid male rabbits, aged between 14 and 15 weeks, were randomly allotted to two groups and housed under either natural day length (n=10, ND) or a constant 16-h daylight exposure of 16 h (n=10, CLD). Other management conditions, such as air temperature or reproductive handling, were identical for both groups. Two successive ejaculates were collected twice weekly from every male, and the first one was used to monitor ejaculate characteristics. Measurement of semen production, in terms of ejaculate and semen volume, sperm concentration and total sperm per ejaculate, and sperm quality, in terms of motility index, viability, morphology and acrosome integrity, was assessed in 783 ejaculates collected during 15 months (from October to December). No differences (P>0.05) in either semen production or sperm quality were shown among ejaculates collected from rabbits housed under ND and CLD conditions. A limited influence of season was observed (P<0.01); semen volume and motility index were highest and lowest, respectively, during summer. The increase of air temperature and humidity index (THI) had a significant detrimental effect (P<0.01) on both sperm production and quality parameters with a lag of 6 and 3 weeks, respectively. On the basis of these findings, annual variations of semen production and sperm quality in male rabbits seems more related to THI than to daylight length under conditions of AI management in the Mediterranean area of Spain.