Influence of wet fixation, staining techniques, and storage time on bull sperm morphology

The influence on sperm morphology of different methods for preparation of semen and of storage in a fixative solution was examined in 27 beef bulls subjected to a regular breeding health examination. Sperm head morphology under light microscopy did not differ between smears of fresh semen stained with carbol-fuchsin-eosin (Williams staining) or Nigrosin-Eosin. Nor was there any difference between samples stained immediately after collection and those stained after 1 month of storage at + 4 degrees C in buffered formal-saline solution. Formol-saline fixed spermatozoa examined in wet preparations under phase contrast microscopy had a higher prevalence of acrosome defects and cytoplasmic droplets than stained smears of fresh semen under light microscopy. One month of storage in formol-saline did not affect the prevalence of acrosome defects or cytoplasmic droplets. There was no influence of fixation method (wet or dry), staining, examination technique, or storage time on midpiece or sperm tail morphology. The affinity of spermatozoa to eosin at staining with Nigrosin-Eosin ("live and dead count") did not differ between fresh semen and spermatozoa that had been stored in formol-saline for 1 month. It is concluded that bull semen can be stored for at least 1 month at + 4 degrees C in buffered formal-saline without major changes in sperm morphology. Furthermore, examination of wet preparations of fixed spermatozoa under phase contrast microscope is likely to yield the most accurate results for morphological characteristics like acrosome morphology and cytoplasmic droplets.     

Assessment of the morphology of frozen-thawed bull sperm in relation to its cryopreservation for artificial insemination

The morphology of sperm in raw semen was compared with that of the live sperm in semen which had been frozen and thawed. The thawed semen was stained with 6-carboxyfluorescein diacetate and propidium iodide and examined by fluorescence microscopy; smears of the raw semen were stained with eosin and nigrosin. Thirty-four ejaculates from 24 bulls of various breeds were examined. There were fewer abnormal heads, detached heads, coiled tails and proximal cytoplasmic droplets/pseudodroplets in the thawed semen than in the raw semen, there was no change in the number of bent tails, but the number of distal cytoplasmic droplets/pseudodroplets increased. There were no significant differences in morphology between ejaculates which passed or failed the osmotic resistance test after thawing, but failed batches tended to have more distal cytoplasmic droplets/pseudodroplets.     

Morphological features of spermatozoa of swamp buffalo AI bulls in Thailand

The appearance and incidence of sperm abnormalities was studied in 115 ejaculates, collected periodically over 1 year covering all seasons from five mature, healthy swamp buffalo (Bubalus bubalis) bulls reared under tropical conditions and serving as the current source of semen for artificial insemination (AI) in Thailand. Light microscopy of stained smears was used to investigate sperm head shape morphology, while unstained wet smears were used to examine other sperm abnormalities. The most commonly found morphological aberrations were pear-shaped spermatozoa, knobbed acrosomes, proximal cytoplasmic droplets, simple bent tails and coiled tails under the head, whose ultrastructure (scanning electron microscopy) corresponded to what has been found in other species of bovidae, including varieties of buffalo. The mean prevalence (as least squares mean +/- SEM) of sperm abnormalities was low (below 15%), corresponding to healthy spermiograms. The younger bulls (<10 years old, n = 3) had less abnormalities than the older ones (10.1 +/- 0.6% versus 14.1 +/- 0.8%, P < 0.001, n = 2), including abnormalities of sperm head shape (1.1 +/- 0.3% versus 3.6 +/- 0.3, P < 0.001), acrosome defects with knobbed acrosomes (1.1 +/- 0.2% versus 1.2 +/- 0.3%, P < 0.001), spermatozoa with proximal cytoplasmic droplets (2.7 +/- 0.1% versus 1.4 +/- 0.2%, P < 0.001), defective mid-pieces (0.2 +/- 0.1% versus 0.3 +/- 0.1%) and abnormal sperm tails (3.1 +/- 0.3% versus 5.7 +/- 0.4%, P < 0.001). The within-bull effect of the year solely affected the incidence of pear-shaped spermatozoa while the incidences of abnormal contour, variable size of sperm head shapes, abnormal mid-piece and simple bent tail among bulls were affected by ejaculate (week of collection). Interaction between age and ejaculate affected only the prevalence of spermatozoa with proximal cytoplasmic droplets. In conclusion, the types of defects encountered were similar to those found in other bovidae, with a very low prevalence over the year the AI sires were followed through.     

Morphology and Chromatin Integrity of Stallion Spermatozoa Prepared by Density Gradient and Single Layer Centrifugation Through Silica Colloids

The objective was to investigate whether it is possible to improve the quality of stallion semen, with respect to sperm morphology and chromatin integrity, both of which have been linked to fertility, using either density gradient centrifugation (DGC) or a new method, hereby named single layer centrifugation (SLC). The two methods of colloidal centrifugation were evaluated using 38 ejaculates from 10 stallions. Sperm morphology, subjective motility and sperm chromatin integrity were compared in uncentrifuged samples and in centrifuged sperm preparations. The proportion of morphologically normal spermatozoa varied between stallions (p < 0.001) and was increased by both methods of colloidal centrifugation (median value before centrifugation 67.5%; after SLC 78%; after DGC 77%; p < 0.001). The incidence of certain abnormalities was reduced, e.g. proximal cytoplasmic droplets were reduced from 12.9% to 8.8% (p < 0.001), and mid-piece defects from 5.3% to 1.4% (p < 0.05). Similarly, sperm motility and chromatin integrity were significantly improved (p < 0.001), with no difference between the two centrifugation methods. Centrifugation through colloids can enrich the proportions of stallion spermatozoa with normal morphology and normal chromatin structure in sperm preparations. The new method, SLC, was as effective as DGC in selecting motile stallion spermatozoa with normal morphology and intact chromatin. SLC, being simpler to use than DGC, would be appropriate for routine use by stud personnel to improve stallion sperm quality in insemination doses.     

Effect of temperature and humidity on sperm morphology in duroc boars under different housing systems in Thailand

The aim of this study was to investigate the effect of season, temperature, humidity, age of the boar, and semen collection interval on sperm morphology in Duroc boars in Thailand, kept either in a conventional open air system (CONV) or in an evaporative cooling system (EVAP). In total, 1176 ejaculates from 110 sexually mature boars in six CONV herds and five EVAP herds were morphologically examined during a one-year period. Analysis of variance was applied to the data. Minor differences in the sperm morphology traits analyzed were found between the housing systems. There was a significant seasonal effect (two-month periods) on the percentage of morphologically normal spermatozoa (normal1), morphologically normal spermatozoa including spermatozoa with distal cytoplasmic droplets (normal2), proximal cytoplasmic droplets (prox), and sperm head abnormalities (P相似文献   

The Proportion of Beef Bulls in Sweden with Mature Spermiograms at 11–13 months of Age

Semen samples collected postmortem from 142 yearling beef bulls (11-13 months old) of three different breeds (Charolais, Hereford and Simmental) were examined to evaluate the proportion of bulls with mature spermiograms. Before slaughter, testes and epididymides were clinically examined and scrotal circumferences were measured. Aliquots of the cauda epididymal contents taken postmortem were used for sperm morphology examination. Sperm head morphology was studied in dry smears stained with carbol-fuchsine. For each preparation, 500 spermatozoa were counted in each smear under light microscope (x 1000). The presence of proximal cytoplasmic droplets, abnormal acrosomes, detached heads and abnormalities of the midpiece and tail were recorded in wet preparations of formol-saline-fixed spermatozoa. For each preparation, 200 spermatozoa were counted in each preparation under a phase-contrast microscope (x 1000). The abnormalities were classified according to a classification system developed by Bane (1961). Morphological abnormalities were recorded as a percentage of the total number of counted spermatozoa. Criteria for a spermiogram to be considered mature included <15% abnormal heads and <15% proximal droplets. According to this definition approximately 48% (68 of 142) of the examined bulls were considered mature. The bulls in this study represent approximately one-fifth of the total amount of performance-tested beef bulls in Sweden during 5 years. Our results indicate that only less than half of the Swedish yearling beef bulls at the testing station appear to have a mature spermiogram at the time they are offered for breeding purposes.     

Genetic parameter estimates for scrotal circumference and semen characteristics of Line 1 Hereford bulls

The objectives of this study were to estimate heritability for scrotal circumference (SC) and semen traits and their genetic correlations (rg) with birth weight (BRW). Semen traits were recorded for Line 1 Hereford bulls (n = 841), born in 1963 or from 1967 to 2000, that were selected for use at Fort Keogh (Miles City, MT) or for sale. Semen was collected by electroejaculation when bulls were a mean age of 446 d. Phenotypes were BRW, SC, ejaculate volume, subjective scores for ejaculate color, swirl, sperm concentration and motility, and percentages of sperm classified as normal and live or having abnormal heads, abnormal midpieces, proximal cytoplasmic droplets (primary abnormalities), bent tails, coiled tails, or distal cytoplasmic droplets (secondary abnormalities). Percentages of primary and secondary also were calculated. Data were analyzed using multiple-trait derivative-free REML. Models included fixed effects for contemporary group, age of dam, age of bull, inbreeding of the bull and his dam, and random animal and residual effects. Random maternal and permanent maternal environmental effects were also included in the model for BRW. Estimates of heritability for BRW, SC, semen color, volume, concentration, swirl, motility, and percentages of normal, live, abnormal heads, abnormal midpieces, proximal cytoplasmic droplets, bent tails, coiled tails, distal cytoplasmic droplets, and primary and secondary abnormalities were 0.34, 0.57, 0.15, 0.09, 0.16, 0.21, 0.22, 0.35, 0.22, 0.00 0.16, 0.37, 0.00 0.34 0.00, 0.30, and 0.33, respectively. Estimates of rg for SC with color, volume, concentration, swirl, motility, and percentages of live, normal, and primary and secondary abnormalities were 0.73, 0.20, 0.77, 0.40, 0.34, 0.63, 0.33, -0.36, and -0.45, respectively. Estimates of rg for BRW with SC, color, volume, concentration, swirl, motility, and percentages live, normal, and primary and secondary abnormalities were 0.28, 0.60, 0.08, 0.58, 0.44, 0.21, 0.34, 0.20, -0.02, and -0.16, respectively. If selection pressure was applied to increase SC, all of the phenotypes evaluated would be expected to improve. Predicted correlated responses in semen characteristics per genetic SD of selection applied to SC were 0.87 genetic SD or less. If selection pressure was applied to reduce BRW, the correlated responses would generally be smaller but antagonistic to improving all of the phenotypes evaluated. Predicted correlated responses in SC and semen characteristics per genetic SD of selection applied to BRW were less than 0.35 genetic SD.     

Evaluation of bovine spermatozoal morphologic features after staining or fixation.

Two experiments were conducted to evaluate effects of 3 stains and 2 fixatives on morphologic features of bovine spermatozoa. In experiment 1, the morphologic features of acrosomes of raw and incubated, extended spermatozoa were evaluated after staining with Hancock's Blom's or Wells-Awa's stains or after fixation with buffered glutaraldehyde. Evaluations were done of stained smears by bright field microscopy and of fixed, unstained preparations, by differential interference contrast microscopy, using wet mounts. Raw semen samples from 1st ejaculates of 80 bulls were evaluated. The percentage of spermatozoa with intact acrosomes averaged 83.5% in unstained preparations fixed in glutaraldehyde, compared with averages of 68.1, 74.5, and 67.4% for smears stained with Hancock's, Blom's, or Wells-Awa's procedures (P less than 0.01). From these results, it appeared that procedures for preparing stained smears were detrimental to acrosomes. Although counts for other acrosomal abnormalities differed (P less than 0.01) in each treatment, patterns were inconsistent. With incubated, extended spermatozoa from 57 bulls, glutaraldehyde-fixed, unstained samples had more (55%) intact acrosomes (P less than 0.01) than did samples stained with Hancock's or Blom's procedures (24.0 and 34.7%, respectively, but the former were not significantly different from Wells-Awa-stained smears (49.3% intact acrosomes). In experiment 2, several morphologic characteristics of spermatozoa from 15 1st ejaculates of 7 bulls were evaluated after staining with Hancock's or Blom's stains or after fixation in buffered glutaraldehyde or buffered formal saline fixatives. Higher counts (P less than 0.01) of head abnormalities were found in wet, unstained fixed preparations (4.83, 4.47, 7.87, and 7.93% respectively, for Hancock's, Blom's, glutaraldehyde, and formol saline methods). There were more (P less than 0.05) separated heads on stained, dry smears (1.43, 1.23, 0.47, and 0.47%, respectively, for Hancock's, Blom's, glutaraldehyde, and formol saline procedures). Fixation with buffered glutaraldehyde resulted in higher counts (P less than 0.01) of proximal protoplasmic droplets (2.47, 1.03, 0.67, and 1.43%, respectively, for glutaraldehyde, Hancock's, Blom's, and formol saline procedures). Although not significant, the same trend was observed for distal protoplasmic droplets...     

Assessment of Sperm Morphology in Zebu Bulls, under Field Conditions in the Tropics

Sperm morphology was studied in 302 extensively managed Zebu bulls (aged 1.5–9 years), classified as sound (n=166) or unsound (n=136) for breeding, under field conditions in the dry tropics of Costa Rica. Single semen samples were collected by electro‐ejaculation and fixed in formol‐saline solution immediately after collection. Sperm morphology was determined in the field on wet smears using a microscope equipped with phase‐contrast optics, and further determined in the laboratory on air‐dried smears stained with carbol‐fuchsin. The frequencies of sperm abnormalities (such as abnormal acrosome, head, neck, mid‐piece, tail, and presence of cytoplasmic droplets) were recorded as a percentage of the total number of counted spermatozoa (400 cells). Zebu bulls considered unsound for breeding showed a higher mean prevalence (p < 0.05) of knobbed acrosomes (4.0 versus 0.9%), head defects [specifically, nuclear invaginations and heads with abnormal shapes and sizes (27.6 versus 4.0%)], abnormal tails (11.2 versus 4.7%), and proximal droplets (8.4 versus 1.6%), compared with bulls considered sound for breeding. In these latter bulls, the abnormality most commonly seen was the presence of single bent tails with an entrapped cytoplasmic droplet (3.0 ± 3.7%). Young Zebu bulls (i.e. bulls under 2 years of age) showed a higher percentage of missing acrosomes, and proximal cytoplasmic droplets, than older sires (12.1 versus 2.4%, and 23.9 versus 3.6%, respectively; p < 0.05), interpreted as an indication of low ejaculation frequency and sexual immaturity, respectively. Bulls with a long scrotum and soft testicular consistency (TC) at palpation showed higher percentages of abnormal sperm heads in the ejaculate than bulls with a normal scrotal length (SL) and a normal TC (32.7 versus 12.8% and 30.7 versus 10.3%, respectively; p < 0.05). In addition, Zebu bulls with a scrotal circumference (SC) ≤ 30 cm showed a higher prevalence of proximal cytoplasmic droplets than bulls whose SC was > 30 cm (9.8 versus 2.6%, p < 0.05). A higher mean percentage of abnormally sized and shaped heads, especially undeveloped and narrow at the base, was more frequently found in stained smears than in unstained samples (26.0 versus 9.9%, p < 0.05), which clearly underlines the importance of using both stained and wet smears when assessing sperm head morphology. However, for a quick assessment of sperm morphology under field, tropical conditions, phase‐contrast microscopy provides useful information for the spermiogramme evaluation.     

Changes in Mithun (Bos frontalis) spermatozoa during epididymal passage

Genital organs of 10 healthy, adult Mithun bulls (6-8 years old) that were slaughtered at the dwellings of tribal people for meat were collected. Immediately after collection, spermatozoa from 3 different regions of the epididymis, i.e. the head, body and tail, were obtained to study morphological changes of the spermatozoa during passage through these regions. The prevalence of proximal cytoplasmic droplets significantly decreased from the head to the tail of the epididymis. Conversely, the percentage of distal cytoplasmic droplets increased significantly from the head to the tail region. The incidence of tailless heads rose significantly from head to body and then reduced significantly in the tail region. The percentage of total head abnormalities did, however, not change markedly, but total mid-piece and tail abnormalities differed significantly between the three epididymal regions.     

A spermatozoal midpiece abnormality associated with infertility in a Llasa Apso dog

The semen of a 9-year-old Llasa Apso dog with a recent history of infertility was examined by light and electron microscopy. The presenting sperm abnormality was a high percentage (87 per cent) of cells with proximal cytoplasmic droplets. With the higher resolution of differential interference contrast microscopy and with transmission electron microscopy (TEM) it was apparent that not all of the midpiece abnormalities had the appearance of typical cytoplasmic droplets. Furthermore, by TEM both axonemal and mitochondrial defects were readily demonstrated, often with evidence of degenerative mitochondria in cytoplasmic swellings at other levels of the midpiece. These severe abnormalities were apparent only by TEM of thin sections, although scanning EM was useful in identifying abnormal midpiece outlines. It is suggested that the failure of droplet migration and the midpiece ultrastructural defects were linked to a common fault in spermateliosis.     

Influence of Temperature during Glycerol Addition and Post-thaw Dilution on the Quality of Canine Frozen Semen

Contents The aim of the present study was to compare the influence of room temperature (27 degrees C) and 4 degrees C during glycerol addition on canine semen cryopreservation and verify the effect of different post-thawing dilutions on canine semen. Ten ejaculates from five stud dogs were collected by digital manipulation. Semen samples were evaluated and further divided into two aliquots. The first aliquot was extended in Tris-egg yolk-glycerol at 27 degrees C and the second one received glycerol at 4 degrees C. Samples were frozen and stored in liquid nitrogen. After 1 week, samples were thawed and submitted to evaluations of progressive sperm motility, morphology, acrosomal integrity, hypo-osmotic swelling (HOST) and thermoresistance tests. For thermoresistance test, aliquots were divided in two portions: one portion was kept undiluted (1 : 0) and the other one was diluted in a 1 : 4 ratio (one part semen to four parts extender). No differences were observed between temperatures for glycerol addition regarding seminal parameters evaluated. Furthermore, post-thawing dilutions demonstrated similar effect on canine semen longevity. Correlations among post-thaw sperm motility and HOST and results from thermoresistance test were observed for both temperatures for glycerol addition. In conclusion, glycerol could be added to canine semen at room temperature (27 degrees C) or at 4 degrees C. Moreover, there is no need to extend canine semen after thawing for the thermoresistance test, but if we need to increase the inseminating volume for artificial inseminations, the addition of extender will not damage the semen.     

Comparison of the effect of various chemical stabilizers and lyophilization cycles on the thermostability of a Vero cell-adapted rinderpest vaccine

The thermostability of a rinderpest vaccine produced on Vero cells was evaluated using a variety of chemical stabilizers and lyophilization protocols. Three stabilizer preparations and three lyophilization schedules were examined using accelerated stability testing at 37 degrees C. The vaccine preparation exhibiting the greatest stability at 37 degrees C was tested at three additional temperatures, 42, 45 and 56 degrees C, and an Arrhenius plot was constructed from the data. The stability of the reconstituted vaccine produced with the two most efficacious stabilizers was examined using three different diluent preparations. The stabilization method and high Vero cell virus batch titers resulted in a lyophilized vaccine which maintained the minimum required dose of log10 2.5 TCID50 tissue culture infectious dose for more than 20 weeks at 37 degrees C.     

Change in Morphology of Spermatozoa from Dismount Semen during the Breeding Season in Thoroughbred Stallions in Japan

To clarify the physiological changes of sperm morphology in active Thoroughbred stallions during the breeding season, we examined the dismount semen collected from the penile urethra immediately after service. The spermatozoa were analyzed for relationships between the morphology and the stallion’s age or the number of services. Seasonal variation was apparent in the rate of the sperm tail abnormalities, spermatozoa with cytoplasmic droplets, appearance of medusa cells, and sperm head length. Area and width of the sperm head correlated negatively with age (P<0.05). The rate of appearance of medusa cells and the length of the sperm head were positively related to the number of services (P<0.05), and the aspect ratio was negatively related (P<0.01).     

Spermatozoa morphology during the breeding season in Thoroughbred stallions in Japan

The morphology of spermatozoa of modern Thoroughbred stallions in Japan was investigated during the breeding season. A total of 299 semen samples were collected from the penises of 16 stallions immediately after service. The rate of abnormalities in sperm heads and tails, spermatozoa with cytoplasmic droplets and slides with medusa cells to total observed slides in each stallion were 3.9 +/- 2.1%, 11.5 +/- 5.9%, 2.4 +/- 2.6% and 20.1%, respectively. The values for the area, length, width and aspect ratio of the stallion sperm head were 12.54 +/- 1.34 microm(2), 5.93 +/- 0.40 microm, 2.69 +/- 0.21 microm and 0.46 +/- 0.05, respectively. With the exception of medusa cells, the features were significantly different among the stallions (P<0.05).     

The Effects of Trypanosomosis on Sperm Morphology in Zebu×Friesian Crossbred Bulls

Detailed studies of sperm morphological abnormalities were carried out on 12 Zebu x Friesian crossbred bulls used in a study of the effects of trypanosomosis. Four bulls were infected with T. vivax, another four with T. congolense, while four served as controls. The infected bulls developed chronic trypanosomosis. All the bulls initially had very low sperm morphological abnormalities that were within acceptable limits for fertile animals. After infection there was a rapid and progressive increase in all sperm abnormalities. Spermatozoa of infected bulls were highly deformed with multiple morphological defects. Mean percentage pre-infection baseline values prior to infection for acrosomal, sperm-head, detached heads, proximal cytoplasmic droplets, distal cytoplasmic droplets, sperm-tail, midpiece and total sperm morphological defects ranged between 0.1 +/- 0.1 for acrosomal and 8.3 +/- 3.2 for total morphological abnormalities in the semen of the bulls. All the infected bulls developed sperm morphological abnormalities of more than a mean of 40.0% from the 4th week after infection until the end of the investigation and were considered unfit for breeding. At 7 weeks post-infection (PI) until the end of the study (12 weeks PI), the controls had a mean of less than 5% sperm morphological defects, while the infected bulls had 100%. Mean percentage values of sperm morphological defects throughout the duration of the investigation for control bulls were low and within the normal range for fertile bulls. These values differed significantly (p<0.001) from the elevated values of the infected bulls. The results show that trypanosomosis due to T. vivax or T. congolense infection can render Zebu x Friesian crossbred bulls unfit for breeding within a very short time. The resultant infertility could be of economic importance in trypanosomosis-endemic sub-Saharan Africa where Zebu x Friesian crossbred bulls are kept.     

The occurrence of paranuclear vacuoles in sheep ruminal epithelium treated with distilled water in vitro and from sheep injected with a large quantity of water into the rumen

In the histological preparation taken after animal death, paranuclear vacuoles (PV) in sheep ruminal epithelium were already present in high percentage (7.5-20.3%). With the incubation of the ruminal mucosae in distilled water (10-15 degrees C) up to 120 min, PV occurrence did not alter. With incubation in NaCl solution, however, PV value decreased with time proportional to NaCl concentration (0.5, 0.85 or 1.5%). In the preparation taken by biopsy, on the other hand, PV were rare in ruminal epithelium (less than 0.3%). With the injection of warm water (10-141/animal), however, PV occurrence in the ruminal epithelium was not affected. PV as cytoplasmic processes of the ruminal Langerhans cells are probably formed by the reaction of these cells to environmental changes in the tissue caused by animal death.     

Effects of age and season on the type and occurrence of sperm abnormalities in Nubian bucks

Effects of age and season on type and occurrence of sperm abnormalities were examined in semen samples collected from 3 groups of Nubian bucks at ages of 4 to 9 months, 10 to 21 months, and 39 to 50 months. The average total percentage of sperm abnormalities at the onset of puberty (141 +/- 4 days) was 64.6 +/- 14.8% (head, 19.5 +/- 13.6%; middle piece, 17.2 +/- 9.3%; and proximal protoplasmic droplets, 14.6 +/- 10.5%), but this improved rapidly and was reduced to 12.5 +/- 7.5% by 8 months of age (head, 1.9 +/- 4.5%; middle piece, 4.6 +/- 2.8%). Further increase in age, at least up to 4 years, did not reveal a significant effect (P less than 0.05) on the type or percentage of total abnormalities. Similar to age, a comparison of data among seasons did not reveal a significant effect on the type or occurrence of sperm abnormalities in 10- to 21-month-old or 39- to 50-month-old bucks. Seemingly, Nubian bucks started producing good quality semen at 8 months of age, and season did not influence sperm abnormalities.     

Estimation of genetic parameters and effects of cytoplasmic line on scrotal circumference and semen quality traits in Angus bulls

The purpose of this study was to estimate the heritability of scrotal circumference (SC) and semen traits, genetic correlations between SC and semen quality traits, and the effect of cytoplasmic line on SC and semen traits. Breeding soundness exam (BSE) data were collected on registered Angus bulls at 4 ranches over 7 yr. The American Angus Association provided historical pedigree information to estimate the effect of cytoplasmic line on SC and semen quality traits. After editing, the evaluated data set contained 1,281 bulls with breeding soundness exam data that traced back to 100 founder dams. Data were analyzed using a 2-trait animal model to obtain heritability, genetic correlation between SC and semen quality traits, as well as the effect of cytoplasmic line as a random effect for SC, percent motility (MOT), percent primary abnormalities (PRIM), percent secondary abnormalities (SEC), and percent total abnormalities (TOT) using multiple-trait derivative-free REML. Fixed effects included source ranch and collection year, and test age was used as a covariate. Estimates of heritability for SC, MOT, PRIM, SEC, and TOT were 0.46, 0.05, 0.27, 0.23, and 0.25, respectively. Genetic correlations between SC and MOT, PRIM, SEC, and TOT were 0.36, -0.19, -0.11, and -0.23, respectively. The proportions of phenotypic variance accounted for by cytoplasmic line for SC, MOT, PRIM, SEC, and TOT were <0.001, 0.013, 0.023, 0.002, and <0.001, respectively. Genetic correlations between SC and semen quality traits were low to moderate and favorable. Cytoplasmic line may have a marginal effect on MOT and PRIM, but is likely not a significant source of variation for SC, SEC, or TOT.     

Effect of dietary selenium and vitamin E on the ultrastructure and ATP concentration of boar spermatozoa, and the efficacy of added sodium selenite in extended semen on sperm motility

Three experiments evaluated the effects of dietary Se and vitamin E on the ultrastructure of spermatozoa, ATP concentration of spermatozoa, and the effects of adding sodium selenite to semen extenders on subsequent sperm motility. The experiment was a 2 x 2 arrangement of treatments in a randomized complete block design. A total of 10 mature boars were fed from weaning to 18 mo of age diets fortified with two levels of supplemental Se (0 or .5 ppm) or vitamin E (0 or 220 IU/kg diet). The nonfortified diets contained .06 ppm Se and 4.4 IU vitamin E/kg. In Exp. 1, the spermatozoa from all boars were examined by electron microscopy. Vitamin E had no effect on structural abnormalities in the spermatozoa. When the low-Se diet was fed the acrosome or nuclei of the spermatozoa was unaffected, but the mitochondria in the tail midpiece were more oval with wider gaps between organelles. The plasma membrane connection to the tail midpiece was not tightly bound as when boars were fed Se. Immature spermatozoa with cytoplasmic droplets were more numerous when boars were fed the low-Se diet, but the occurrence of midpiece abnormalities occurred in boars fed diets with or without Se or vitamin E. Our results suggest that Se may enhance spermatozoa maturation in the epididymis and may reduce the number of sperm with cytoplasmic droplets. In Exp. 2, the concentration of ATP in the spermatozoa was evaluated in the semen of all treatment boars. When the low-Se diet was fed, ATP concentration was lower (P < .01), whereas vitamin E had no effect on ATP concentration. Experiment 3 investigated the effect of diluting boar semen with a semen extender with sodium selenite added at 0, .3, .6, or .9 ppm Se. Three ejaculates from each boar were used to evaluate these effects on sperm motility to 48 h after dilution. Sperm motility declined (P < .01) when Se was added to the extender, and this decline was exacerbated as the concentration of added Se increased (P < .01). The added Se was demonstrated to be tightly adhered to the spermatozoa. Overall, these results suggest that low Se-diets fed to boars resulted in abnormal spermatozoal mitochondria, a lower ATP concentration in the spermatozoa, and a loose apposition of the plasma membrane to the helical coil of the tail midpiece, but no effect from inadequate vitamin E was demonstrated. Adding sodium selenite to the semen extender reduced sperm cell motility.