Retrograde flow of spermatozoa into the urinary bladder of dogs during ejaculation or after sedation with xylazine

Retrograde flow of spermatozoa into the urinary bladder of dogs during ejaculation or after administration of xylazine was examined. In experiment 1, the mean (+/- SD) spermatozoal concentration in urine collected by cystocentesis before ejaculation was 0.322 +/- 0.645 X 10(6)/ml. After ejaculation, motile spermatozoa were present in the urine collected by cystocentesis from 12 of 15 dogs, and the concentration of spermatozoa in the urine (5.139 +/- 7.014 X 10(6)/ml) was higher (P less than 0.025) than the concentration in the urine collected before ejaculation. The percentage of the total number of spermatozoa that were displaced during ejaculation and flowed into the urinary bladder (retrograde flow) ranged from 0 to 99.75% (24.67 +/- 33.98%). In experiments 2 and 3, administration of xylazine to sexually rested dogs induced retrograde flow of spermatozoa into the urinary bladder. In experiment 2, all dogs had spermatozoa in urine collected after xylazine administration, with motile spermatozoa present in the urine from 9 of 10 dogs. In experiment 3, urine collected from dogs before administration of xylazine was azoospermic or contained few, nonmotile spermatozoa (0.063 +/- 0.135 X 10(6)/ml), whereas urine collected after administration of xylazine had more (P less than 0.025) and motile spermatozoa (3.717 +/- 4.273 X 10(6)/ml). In experiment 4, administration of xylazine to dogs after ejaculation did not increase the concentration of spermatozoa in the urine. Results indicate that spermatozoa flow into the urinary bladder of dogs during ejaculation or after administration of xylazine to sexually rested dogs.     

Effect of method of seminal collection on the retrograde flow of spermatozoa into the urinary bladder of rams

The effects of method of seminal collection and a diuretic on retrograde flow of spermatozoa into the urinary bladder of rams were examined. In experiment 1, semen and urine were collected from 8 rams during the non-breeding season. Prior to seminal collection, all rams were given furosemide and a sample of urine was obtained during micturition. Semen was then collected from each ram with an artificial vagina or by electroejaculation in alternate weeks for 4 weeks, and the urine released during the first postseminal collection micturition was collected in 4 consecutive samples. The volume of electroejaculates was larger (P less than 0.0001) than the volume of ejaculates, but the total number of spermatozoa in the electroejaculate or in the ejaculate were not different (P greater than 0.1). Urine obtained before seminal collection was azoospermic or contained few, nonmotile spermatozoa (mean +/- SD = 0.053 +/- 0.114 x 10(6)/ml). The adjusted spermatozoal concentration (mean +/- SD = 1.630 +/- 2.258 x 10(6)/ml) in the urine collected after seminal collection was 31 times higher (P less than 0.0001) and there were motile spermatozoa in most (97%) of the samples. The spermatozoal concentration in sequential samples of urine was not different (P greater than 0.1) between samples and was not affected (P greater than 0.1) by the method of seminal collection. There was a trend, approaching significance (P = 0.052), for an effect of method of seminal collection on the percentage of retrograde flow.(ABSTRACT TRUNCATED AT 250 WORDS)     

Retrograde flow of spermatozoa into the urinary bladder of rams

Retrograde flow of spermatozoa into the urinary bladder of rams during electroejaculation (EE) was examined. In experiment 1, semen and 4 consecutive samples of the urine released during the first post-EE micturition were collected once a week from 6 rams for 5 weeks during the nonbreeding season. The overall mean concentration per milliliter and the mean total number of spermatozoa in the urine ranged from 3.06 to 4.32 X 10(6) and from 80 to 2,865 X 10(6), respectively. The spermatozoal concentration in sequential urine samples was not different between samples, indicating that these spermatozoa had mixed with the urine before micturition. The percentage of the total number of spermatozoa displaced during EE, which flowed into the urinary bladder (retrograde flow), varied among rams (range 3.9% to 80%). The overall mean percentage of retrograde flow during the nonbreeding season was 28.3%. In experiment 2, a catheter was implanted into the urinary bladder of 6 rams (4 rams were from experiment 1), and semen was collected over 4 weeks during the subsequent breeding season. A urine sample was collected from the implanted catheter before EE. Immediately after semen collection, urine was collected by evacuating the bladder. The spermatozoal concentration in the pre-EE urine ranged from 0 to 1.3 X 10(6) (mean +/- SD, 0.17 +/- 0.38 X 10(6)) and was significantly (P less than 0.0001) lower than the spermatozoal concentration in the post-EE urine, which ranged from 1.10 to 22.55 X 10(6) (mean +/- SD, 9.46 +/- 11.30 X 10(6)).(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison between the Sperm Morphology in Semen Samples Obtained from Yearling Beef Bulls by Transrectal Massage of the Ampullae and Cauda Epididymal Dissection

As electroejaculation (EEJ) is prohibited for use on unanaesthetized animals in Sweden, there is a need for an alternative method of semen collection from bulls in the field. The aim of the present study was to evaluate the use of transrectal massage (TM) of the ampullae to collect semen from yearling beef bulls under field conditions in Sweden. Transrectal massage was performed on 52 yearling beef bulls. Volume of semen collected, duration of procedure, percentage progressively motile sperm, and sperm concentration were measured. Smears were prepared for sperm morphology examination. Semen samples were obtained from 47 of 52 bulls. Mean volume was 3.2 ml (SD +/- 3.7), mean duration of collection was 7.4 min (SD +/- 2.8), mean percentage progressively motile sperm was 43.5% (SD +/- 29.2) and mean concentration was 201.9 x 10(6) spermatozoa/ml (SD +/- 278 x 10(6)). Twenty-three of the 52 bulls were slaughtered 3-4 days after semen collection and aliquots of the cauda epididymal contents were collected for sperm morphology examination. The percentages of proximal droplets, abnormal tails and abnormal midpieces were significantly higher (p < 0.05) and the percentage of detached heads was significantly lower (p < 0.05) in the post-mortem samples compared with those in the TM samples. However, importantly there was no significant difference between the two sample types in the percentages of abnormal heads. This study demonstrates that semen can be collected from yearling beef bulls by TM. We think that TM constitutes a useful tool, when semen collection with EEJ or artificial vagina (AV) is not possible under field conditions, when included in the bull breeding soundness evaluation (BBSE) protocol. However, further studies are needed, and presently being carried out, to evaluate if semen samples collected by TM are comparable with semen samples collected by AV.     

Hysteroscopic insemination of low numbers of flow sorted fresh and frozen/thawed stallion spermatozoa

The objective of this experiment was to determine the effects of flow cytometric sorting and freezing on stallion sperm fertility. A 2 x 2 factorial design was used to delineate effects of flow sorting and freezing spermatozoa. Oestrus was synchronised (July-August) in 41 mares by administering 10 ml altrenogest (2.2 mg/ml) per os for 10 consecutive days, followed by 250 microg cloprostenol i.m. on Day 11. Ovulation was induced by administering 3,000 iu hCG i.v. either 6 h (fresh spermatozoa) or 30 h (frozen/thawed spermatozoa) prior to insemination. Mares were assigned randomly to one of 4 sperm treatment groups. Semen was collected from 2 stallions with an artificial vagina and processed for each treatment. Treatment 1 (n = 10 mare cycles) consisted of fresh, nonsorted spermatozoa and Treatment 2 (n = 16 mare cycles) of fresh, flow sorted spermatozoa. Spermatozoa to be sorted were stained with Hoechst 33342 and sorted into X- and Y-chromosome-bearing populations based on DNA content using an SX MoFlo sperm sorter. Treatment 3 (n = 16 mare cycles) consisted of frozen/thawed nonsorted spermatozoa (frozen at 33.5 x 106 sperm/ml in 0.25 ml straws) and Treatment 4 (n = 15 mare cycles) of flow sorted frozen/thawed spermatozoa (frozen at 64.4 x 10(6) sperm/ml). Concentrations of sperm in both cryopreserved treatments were adjusted, based on predetermined average post-thaw motilities, so that each insemination contained approximately 5 x 10(6) motile spermatozoa. Hysteroscopic insemination of 5 x 10(6) motile spermatozoa in a volume of 230 microd was used for all treatments. Pregnancy was determined ultrasonographically 16 days postovulation. No differences were found (P>0.1) in the pregnancy rates for mares inseminated with fresh nonsorted (4/10 = 40.0%), fresh flow sorted (6/16 = 37.5%), frozen/thawed nonsorted (6/16 = 37.5%) and flow sorted frozen/thawed spermatozoa (2/15 = 133%). Pregnancy rates tended (P = 0.12) to be lower following insemination of frozen/thawed flow sorted spermatozoa. Further studies are needed with a larger number of mares to determine if fertility of flow sorted frozen/thawed spermatozoa can be improved.     

Electroejaculation and semen cryopreservation of free-ranging Japanese black bears (Ursus thibetanus japonicus)

The Japanese black bear (Ursus thibetanus japonicus) is endangered for extinction in some areas of Japan, and semen collection and cryopreservation are an important means to preserve genetic resources. The aim of this study was to characterize and cryopreserve semen of free-ranging Japanese black bears. Semen was collected by electroejaculation procedure from 4 free-ranging Japanese black bears at the capture point in the field. Ejaculates containing motile sperm were recovered from all of the animals and ejaculate volume, total sperm count, % motility (percentage of motile spermatozoa), % viability (percentage of spermatozoa that excluded eosin) and % abnormal morphology (range (mean)) were 0.65-2.20 (1.51) ml, 99-1082 (490) x 10(6), 5-100 (31), 42-97 (66) and 20-87 (53), respectively. Three of the 4 ejaculates were diluted with an egg yolk-TRIS-citrate-glucose extender and cryopreserved in liquid nitrogen. Motile spermatozoa were observed after freezing and thawing in all cases. This study showed that electroejaculation would be a useful method for collecting semen from free-ranging Japanese black bears and that at least motile spermatozoa would be obtained by freezing the thus collected electroejaculates.     

Evaluation of semen collected by electroejaculation from captive lesser Malay chevrotain (Tragulus javanicus).

Thirteen sexually mature captive male lesser Malay chevrotains (Tragulus javanicus) were each anesthetized twice with tiletamine-zolazepam for electroejaculation. Viable spermatozoa were collected from all animals. The semen was creamy, milky, pale yellowish, or watery. The mean values for ejaculate volume, sperm concentration, and percentages of sperm motility, normality and viability were 23.7 +/- 2.5 microl, 366.9 +/- 127.8 x 10(6) spermatozoa/ml, 40.0% +/- 3.1%, 71.4% +/- 1.6%, and 59.6% +/- 2.1%, respectively. Semen pH was 7-8. No adverse effects of electroejaculation were noted. These are the first reported values for semen of lesser Malay chevrotain. Electroejaculation should be usable for routine semen collection in this species.     

Twenty-four hour urinary protein loss in healthy cats and the urinary protein-creatinine ratio as an estimate

Urinary protein loss was determined in 12 healthy cats. Voided urine was collected and protein quantitated by the Coomassie blue method. Mean protein loss for all cats was 12.65 mg/kg/24 h (5.45 SD). Protein loss for male cats (n = 6) was 16.62 mg/kg/24 h (3.3 SD), which was significantly different (P less than 0.01) from 8.69 mg/kg/24 h (4.09 SD) for females (n = 6). A single urine protein-creatinine ratio correlated well with the total urinary protein loss in mg/kg/24 h. The correlation coefficient for the protein-creatinine ratio in voided urine (UPCV) vs 24-hour urinary protein (UP-24) loss was 0.968, and that for the protein-creatinine ratio in urine obtained by cystocentesis (UPCC) vs UP-24 was 0.945. The regression equations were UPCV = 0.02145 + 0.02338 x UP-24 (mg/kg), and UPCC = 0.02667 + 0.02133 x UP-24 (mg/kg). Using the mean value plus 3 SD of urinary protein loss from the healthy cats in this study, a healthy cat would be expected to have a urinary protein loss of less than 29 mg/kg/24 h. A protein-creatinine ratio from a single urine sample provides an accurate estimate of urinary protein loss in healthy cats.     

Results of analyses and bacterial cultures of urine specimens obtained from clinically normal cats by three methods

Six urine specimens were obtained from each of 6 male and 6 female cats in a 3-week period. The first and last specimens from each cat were obtained by cystocentesis, 2 were obtained by urethral catheterization, and 2 were caught during voiding stimulated by manual bladder compression. Quantitative urine cultures did not reveal bacteriuria in specimens obtained by cystocentesis, and urinary tract infection did not develop during the study. Bacteria were found in 3 (25%) of 12 urine specimens obtained by catheterization of males and in 1 (8%) of 12 specimens obtained by catheterization of females. Magnitude of bacteriuria in specimens obtained by catheterization was 10 to 1,000 organisms/ml. Bacteria also were found in specimens obtained during voiding. Each of 11 (100%) cleanly caught specimens obtained from males and 7 (58%) of 12 voided specimens from females contained bacteria. Magnitude of bacteriuria in voided specimens was usually 100 to 10,000 organisms/ml, but 3 voided specimens contained greater than 10,000 organisms/ml. Many bacteriuric specimens contained more than 1 type of organism; however, bacteria that might be suspected of causing urinary tract infections in cats were found frequently. Urinalyses were performed on 66 specimens. Completeness of urinalyses depended on the volume of specimens available, but results were normal except for evidence of hematuria in a few specimens obtained by cystocentesis or catheterization. Hematuria was usually mild and was attributed to hemorrhage caused by minor urinary tract trauma during urine collection.     

Occurrence of occult bacteriuria in healthy cats

Knowledge of the occurrence of bacteriuria in adult, healthy cats is scarce in the scientific literature. A study was designed to investigate the occurrence of bacteriuria in healthy cats without current or previous signs of lower urinary tract disease. The study included 108 cats, 53 males (49.5%) and 55 females (50.5%). The cats ranged in age between 7 months and 18 years, with a mean age of 4.4 years and a median age of 4.0 years. Urine was obtained by cystocentesis from all the cats, and was submitted for bacteriological analyses. Urine and urine sediment was cultured on separate blood agar plates for quantification and species identification by standard procedures. Detection of ≥10(3)colony forming units (cfu) per ml urine was defined as significant bacteriuria. Significant bacteriuria exceeding 10(5) cfu/ml was detected in one sample with a combination of Enterococcus species and Staphylococcus species. There was no bacterial growth in the urine samples from 107 cats (99.1%). Results from our study indicate that the prevalence of bacteriuria in clinically healthy, adult cats is low. Also, that contamination of samples is rare when urine is collected by cystocentesis.     

Xylazine does not induce retrograde flow of spermatozoa into the urinary bladder of sexually rested boars

The effect of xylazine on the retrograde flow of spermatozoa from their storage sites in the epididymides and vasa deferentia into the urinary bladder of sexually rested boars was examined. The bladder of four boars was evacuated through a surgically implanted urinary catheter and the urine was examined for the presence of spermatozoa. Boars were then given an injection of 2.2 mg of xylazine per kilogram of body weight and, immediately thereafter, 500 ml of saline was infused into the urinary bladder. Approximately 50 ml of the post-treatment mixture of urine and saline, referred to as 'urine', was collected through the catheter at 5, 10, 15, 20, 30 and 45 min after the injection of xylazine, and examined immediately for the presence and motility of spermatozoa. At 60 min, the urinary bladder was evacuated and the remaining 'urine' was examined for the presence and motility of spermatozoa. None of the pre-xylazine urine and post-xylazine fractions of 'urine' had motile spermatozoa and xylazine did not increase (P > 0.1) the concentration and the number of spermatozoa in the post-treatment 'urine'. Thus, in contrast to other species, xylazine does not induce retrograde flow of spermatozoa into the urinary bladder of boars.     

Yohimbine prevents the retrograde flow of spermatozoa into the urinary bladder of dogs induced by xylazine

This study was carried out to determine whether yohimbine antagonizes the retrograde flow of spermatozoa into the urinary bladder of dogs caused by xylazine. Adult dogs were assigned to one of four groups of six dogs each and treated as follows: saline control, xylazine (2.2 mg/kg, i.m.), yohimbine (0.2 mg/kg, im.), yohimbine/xylazine (yohimbine, 0.2 mg/kg, i.m., followed 10 min later by xylazine. 2.2 mg/kg, i.m.). Pre- and post-treatment urine were collected by cystocentesis from all dogs. The mean (± SD) adjusted total number of spermatozoa in the post-treatment urine of xylazine-treated dogs (141.02 ± 136.75 × 10⁶) was 15 times higher ( P < 0.05) than the number in the post-treatment urine of control dogs (9.16 ± 20.26 × 10⁶), 1763 times higher ( P < 0.05) than the number in the urine of yohimbine-treated dogs (0.08 ± 0.20 × 10⁶), and 56 times higher ( P < 0.05) than the total number in the post-treatment urine of yohimbine/xylazine-treated dogs (2.54 ± 4.54 × 10⁶). These results confirm that xylazine induces a significant ( P = 0.007) displacement of spermatozoa into the urinary bladder of dogs and demonstrate that pre-treatment with yohimbine prevents this effect.     

Influence of dietary source of phosphorus on fecal and urinary excretion of phosphorus and other minerals by male cats

Twelve male cats were fed 2 diets that differed in the source of P. In diet 1 (1.4% P), 62.7% of P originated from poultry, meat, and fish meal, and the remainder from other organic ingredients of food. In diet 2 (1.6% P), 63.5% of P was derived from neutral monobasic/dibasic salts, and the remainder from other organic ingredients of the food. The P intake was nearly the same with both diets, but there was a significant (P less than 0.05) difference between diets in the percentage of ingested P that was excreted in the urine (14.7 +/- 5.3% for diet 1; 34.9 +/- 8.4% for diet 2), and in 6-day urinary P excretion (774 +/- 290 mg for diet 1; 2,004 +/- 556 mg for diet 2). The P concentrations in urine samples obtained by cystocentesis after cats ate were significantly (P less than 0.05) higher when cats were fed diet 2 than when those same cats were fed diet 1. Plasma P concentrations increased after ingestion of diet 2, but were unchanged after ingestion of diet 1. Seemingly, urinary excretion of P was markedly influenced by dietary composition. Diets with the same P content have potential for different biologic effects because of differences in availability of P.     

Semen collection in rhinoceroses (Rhinoceros unicornis, Diceros bicornis, Ceratotherium simum) by electroejaculation with a uniquely designed probe.

Electroejaculation in rhinoceroses has historically yielded inconsistent results, with the collection of high-quality, sperm-rich samples rare. The goal of this study was to develop a reliable method of electroejaculation in the rhinoceros by designing a rectal probe that appropriately fits the anatomy of this taxon and refining the procedure. A curved probe handle ending in an oblate, ellipsoid head was built using readily available supplies. A combination of rectal massage, penile massage, and electrical stimulation with a specially designed probe was employed in attempts to collect semen on 14 occasions from greater one-horned rhinoceroses (Rhinoceros unicornis; n = 4), black rhinoceroses (Diceros bicornis; n = 2) and a southern white rhinoceros (Ceratotherium simum; n = 1). During 13 of the 14 attempts, ejaculates were collected in multiple fractions. All but one of the ejaculates contained spermatozoa, and seven ejaculates contained good-quality fractions of semen (-60% sperm motility; > or =20 x 106 spermatozoa/ml) suitable for sperm banking and assisted reproduction procedures. Mean (+/-SEM) values for volume, pH, osmolality, and total sperm number for ejaculates containing good-quality fractions (98.2 +/-21.8 ml, 8.5+/-0.1, 290.4+/-6.7 mOsm, and 37.1+/-12.0 x 10(9), respectively) did not differ (P > 0.05) from those containing only poor-quality samples. Urine and/or erythrocyte contamination was not uncommon in fractions of both ejaculate types. Males producing good-quality samples ranged in age from 7 to 34 yr. None of the samples contained > or =75% morphologically normal spermatozoa. Electroejaculation with a uniquely designed probe consistently produced ejaculates in the rhinoceros. However, the production of high-quality samples continued to be challenging, occurring in only 50% of collection attempts. Regardless, the technology has progressed to a stage at which good-quality semen samples can be produced for sperm banking and assisted reproduction, and thereby can be integrated into intensive rhinoceros management strategies for the ultimate survival of this taxon.     

Effect of method of collection on seminal characteristics of the domestic cat

The effect of level of voltage and method of collection on seminal characteristics were studied in the domestic cat. A Latin square design was used to determine the effects of voltage (1, 2, 4, or 6 V) on seminal characteristics of electroejaculates for 8 cats (experiment 1). There was a significant effect of cat on the total volume (P less than 0.005), number of spermatozoa (P less than 0.05), pH (P less than 0.05), and osmolality (P less than 0.025). There was an effect of week for the pH (P less than 0.05) and osmolality (P less than 0.005) of semen. Voltage of stimulation affected the total volume (P less than 0.005), total number of spermatozoa (P less than 0.025), and osmolality (P less than 0.005) of semen. There were trends (P less than 0.1) for an effect of cat and an effect of voltage on spermatozoal motility. Urine contamination was not observed in any of the electroejaculates. A 2, 2 X 2 Latin square design was used in experiment 2 to determine the effect of method of collection (artificial vagina or electroejaculation) on seminal characteristics for 4 cats. Electroejaculation was performed, using the 6-V electrical stimulus selected from experiment 1. There was a significant (P less than 0.025) effect of cat on the motility of the spermatozoa in the viability preparation and a trend (P less than 0.1) for an effect of cat on spermatozoal viability.(ABSTRACT TRUNCATED AT 250 WORDS)     

Distribution of Spermatozoa and Embryos in the Female Reproductive Tract after Unilateral Deep Intra Uterine Insemination in the Pig

The present study was performed to investigate the number of either the spermatozoa or the embryos in the reproductive tracts of sows after unilateral, deep, intra uterine insemination (DIUI). Two experiments were conducted, 10 sows were used in experiment I and eight sows were used in experiment II. Transrectal ultrasonography was used to examine the time when ovulation took place in relation to oestrus behaviour. The sows were inseminated with a single dose of diluted fresh semen 6-8 h prior to expected ovulation, during the second oestrus after weaning. In experimental I, five sows were inseminated by a conventional artificial insemination (AI) technique using 100 ml of diluted fresh semen, containing 3000 x 10(6) motile spermatozoa and five sows were inseminated by the DIUI technique with 5 ml of diluted fresh semen, containing 150 x 10(6) motile spermatozoa. The sows were anesthetized and ovario-hysterectomized approximately 24 h after insemination. The oviducts and the uterine horns on each side of the reproductive tracts were divided into seven segments, namely ampulla, cranial isthmus, caudal isthmus, utero-tubal junction (UTJ), cranial uterine horn, middle uterine horn and caudal uterine horn. Each segment of the reproductive tracts was flushed with Beltsville thawing solution (BTS) through the lumen. The total number of spermatozoa in the flushing from each segment were determined. In experimental II, eight sows were inseminated by the DIUI technique using 5.0 ml diluted fresh semen containing 150 x 10(6) motile spermatozoa. The sows were anesthetized 61.1 +/- 12 h after insemination (48-72 h) and the embryos were flushed from the oviduct through the proximal part of the uterine horn. It was revealed that, in experimental I, the spermatozoa were recovered from both sides of the reproductive tract in the AI-group, and from unilateral side of the reproductive tract in the DIUI-group (three sows from the left and two sows from the right sides). The number of spermatozoa recovered from the reproductive tracts was higher in the AI- than the DIUI-group (p < 0.001). In experiment II, fertilization occurred in five of eight sows (62.5%) after DIUI. The number of ova that ovulated were 16.4 +/- 2.6 per sow and the embryos numbering 11.4 +/- 2.3 per sow were recovered from both sides of the reproductive tract. In conclusion, the spermatozoa given by DIUI could be recovered from only one side of the reproductive tract of sows at approximately 24 h after DIUI via the flushing technique. However, embryos were found in both sides of the oviducts and the proximal part of the uterine horns 48-72 h after insemination, indicating that the fertilization occurred in both sides of the oviducts.     

Characterisation of movement pattern and velocities of stallion spermatozoa depending on donor, season and cryopreservation

The aim of the study was to compare different types of movement pattern and velocities of stallion spermatozoa depending on cryopreservation during breeding and non-breeding season. Ejaculates were collected from four stallions during May (n = 24) and December (n = 24). Parameters of sperm movement were evaluated by computer-aided sperm analysis (CASA) system, and included percentages of motile spermatozoa, different patterns of motility, the velocity, linearity (LIN), amplitude of lateral head displacement (ALH) and beat-cross frequency (BCF). In winter the average percentages of motility were slightly higher compared to the breeding season in May (70.8 +/- 12.7% vs. 66.8 +/- 12.2%, respectively). Cryopreservation and thawing led to a significant decrease in the number of motile sperm to 11.3 +/- 5.8% in May and 15.6 +/- 7.0% in December. The pattern of motility was also changed. Detailed analysis by CASA demonstrated that cryopreservation resulted in a shift from the proportions of linear to more non-linear motile spermatozoa and to a significant increase of local motile and hyperactivated spermatozoa. Mean velocity of fresh motile spermatozoa differed between May and December (119.1 +/- 43.9 vs. 164.4 +/- 66.4 microm/sec, respectively; P < 0.05). Cryopreservation and thawing led to a slight increase of curvilinear velocity (VCL) and straight line velocity (VSL). The motility analysis has shown that the parameters BCF and ALH were highly correlated in stallion spermatozoa (r = -0.67; P < 0.001). The BCF of stallion spermatozoa was slightly reduced in the non-breeding season. Altogether, the influence of factors on the motility of stallion spermatozoa has the following rank order: cryopreservation (P < 0.0001) > stallion (P < 0.001) > season (P < 0.05).     

Hyperactivation and acrosome reaction in vitro in spermatozoa ejaculated by cryptorchid dogs after orchiopexy.

Orchiopexy of the cryptorchid (CR) testis and castration of the scrotal testis were performed in three unilaterally CR beagles at six months of age. Induction rates for ejaculated sperm hyperactivation (HA) and the acrosome reaction (AR) in vitro in these orchiopexied dogs were compared with five those in normal beagles one year later. Canine spermatozoa were incubated for 9 hr at 38 degrees C under 5% CO2 in air in canine capacitation medium at a concentration of 30 x 10(6) sperm/ml. HA was observed using high-speed videomicrography. The AR spermatozoa were evaluated by the triple stain technique. As a result, there was no significant difference between 'the CR dogs after orchiopexy' (CDO) and the normal dogs (ND) with respect to sperm motility just after ejaculation. However, sperm motility of CDO decreased markedly during incubation. There was a significant difference in sperm motility between CDO (Mean +/- SD; 47 +/- 12%) and ND (80 +/- 9%) after three hours of incubation (p less than 0.01). No significant difference was observed between CDO and ND with respect to the HA rate of motile spermatozoa throughout the incubation period. The peak of HA rate was found in both CDO (58 +/- 5%) and ND (61 +/- 16%) after seven hours of incubation. The AR rate of spermatozoa in CDO was lower than that in ND after six hours of incubation. The AR rate of CDO (26 +/- 4%) was significantly lower than ND (46 +/- 5%) after eight hours of incubation (p less than 0.01). It is assumed that there might be relation between a rapid decrease of motility and low AR rate in spermatozoa of CDO during incubation.     

Successful Low Dose Insemination of Flow Cytometrically Sorted Ram Spermatozoa in Sheep

The fertility of ram spermatozoa that had undergone flow cytometric sorting (MoFlo SX) and cryopreservation was assessed after low-dose insemination of synchronized Merino ewes. Oestrus was synchronized with progestagen-impregnated pessaries, PMSG and GnRH treatment. Ewes (n = 360) were inseminated with 1 x 10(6), 5 x 10(6) or 15 x 10(6) motile sorted frozen-thawed (S(1), S(5), or S(15) respectively) or non-sorted frozen-thawed (C(1), C(5) or C(15) respectively) spermatozoa from three rams. An additional group of ewes were inseminated with 50 x 10(6) motile non-sorted frozen-thawed spermatozoa (C(50)) to provide a commercial dose control. The percentage of ewes lambing after insemination was similar for C(50) (24/38, 63.2%), C(15) (37/54, 68.5%), S(15) (38/57, 66.7%), S(5) (37/56, 66.1%) and S(1) (32/52, 61.5%) groups (p > 0.05), but lower for C(5) (19/48, 39.6%) and C(1) (19/55, 34.5%) treatments (p < 0.05). This study demonstrates sorted ram spermatozoa are equally fertile to non-sorted spermatozoa even when inseminated at 2% of the dose. Furthermore, at very low artificial insemination doses (1 or 5 million motile) the fertility of sorted ram spermatozoa is superior to non-sorted spermatozoa inseminated in equal numbers. These results have significance for the future commercialization of sex-preselection technology in sheep as a reduction in the minimum effective sperm number will allow a corresponding decrease in the associated cost per dose.     

Sperm Morphology in the Domestic Cat, and its Relation with Fertility: A Retrospective Study

Knowledge about normal ranges in semen quality and the association between sperm morphology and fertility in felids is limited. The aims of this retrospective study were to (1) define a normal spermiogram in cats; (2) evaluate possible effects of season, age and breed on sperm morphology; and (3) evaluate the relationship between sperm morphology and fertility. Semen samples collected by electroejaculation from 52 cats were evaluated for sperm morphology. The cats constituted two groups: a general population of cats (n = 48) and cats examined because of poor breeding records (n = 4). The general population was divided into household (n = 20), pedigree (n = 19) and colony cats (n = 9) and into three age classes, <12 months, 12-59 months and >or=60 months. The median percentage of normal spermatozoa in the general population was 44.0% (range 1.0-91.0%). Criteria were tentatively set for what was considered a normal spermiogram. The mean percentage of normal spermatozoa was higher during February to July than during August to January (p < 0.05). Pedigree cats had a lower mean percentage of normal spermatozoa than did household cats (p < 0.05). Age had no effect on the percentage of normal spermatozoa but was positively correlated with the percentage of proximal droplets. Of the cats with <40% normal spermatozoa (n = 19), all those with known breeding records (n = 11) had produced litters. The four cats examined because of poor breeding results had higher percentages of different sperm abnormalities than tentatively stipulated for the normal spermiogram. In two of these cats both sperm morphology and fertility changed over time.