Assessment of acrosomal integrity of vervet monkey spermatozoa after cryopreservation.

Cryopreservation of Vervet monkey semen caused a highly significant reduction in the percentage of normal acrosomes and a highly significant increase in the percentage of mildly damaged, severely damaged, and lost acrosomes. This was demonstrated by staining ten post-thaw semen samples, which enabled a visualization of the acrosomal damage. The incidence of post-thaw intact acrosomes in this cryopreservation method was found to be similar to that reported for man and cynomolgus monkey semen.

Cryopreservation of vervet monkey semen and recovery of progressively motile spermatozoa.

A method to cryopreserve semen from the Vervet monkey (Cercopithecus aethiops) has been developed, yielding a mean post thaw recovery of 63.60% of pre-freeze progressive motility. The extender contained a TES-TRIS buffer, egg yolk, dextrose, streptomycin, penicillin, and glycerol as cryoprotectant. The pH ranged from 7.10-7.18 and the osmolality was 330-345 mOsmol/L.

Sperm morphology and fertility in the dog.

Normal and abnormal dog spermatozoa were studied by means of bright field light and transmission electron microscopy and the effect of abnormal spermatozoa on dog fertility was evaluated. Semen samples were collected by digital manipulation from 67 dogs of 34 different breeds. Ten of the samples were examined ultrastructurally. The association between the percentage abnormal spermatozoa in the ejaculate and the fertility of the dog was statistically evaluated, based on insemination of 42 bitches. On this basis, the male dogs were divided into normal and subnormal groups. The percentage normal morphology below which fertility was adversely affected was found to be 60%: the fertility of dogs with > 60% normal morphology was 61% (14 of 23 inseminated bitches) whereas the fertility of dogs with < 60% normal morphology was 13% (two of 15 inseminated bitches). There was no statistical difference between the ages of the dogs in the two groups; from this it was concluded that subfertility may affect a dog at any age.

Ultrastructural parameters of fertile cryopreserved human sperm.

Two hundred and forty-four cryopreserved semen samples were used for artificial insemination by donor (AID). All samples were examined for motility and concentration. Thirty of the samples resulted in pregnancies. These samples were further examined ultrastructurally. There was no difference in sperm motility or concentration between the samples that did or did not result in a pregnancy. The ultrastructural characteristics of the samples that resulted in pregnancies revealed that only 15% of sperm (SD = 7.67) possessed normal morphology and had undamaged acrosomes after cryopreservation.

The embryo freezing programme at Groote Schuur Hospital. First results.

'Surplus' embryos from 28 patients on the in vitro fertilisation programme at Groote Schuur Hospital were frozen for transfer in later cycles. Depending on the stage of the embryos, either propanediol or glycerol was used as a cryoprotectant. In 20 patients embryos survived the thawing procedure and were transferred. Five pregnancies resulted, 2 twin and 3 singleton. The pregnancy rate per embryo transfer in this group of patients in thus 25%. The pregnancy rate per freezing procedure is 19.2%.

An outbreak of granulomatous vulvitis in feedlot heifers.

An unusual form of vulvitis occurred in outbreak form affecting at least 18 of a group of 35 to 40 heifers soon after their introduction to a feedlot in the western Transvaal area of the Republic of South Africa. Initially, affected animals showed a knob-like swelling of the ventral commissure of the vulva. This grew in size up to approximately 6 cm in diameter and then appeared to burst open and give rise to a granulomatous lesion. Bovine herpesvirus type-1, Moraxella bovis, Mycoplasma bovigenitalium and Mycoplasma bovis were isolated from 2 affected heifers referred for diagnostic investigation. Mycoplasma canadense was isolated from one of the heifers. Attempts to reproduce the disease by transferring an excised portion of the lesion to unaffected animals, resulted only in lesions typical of uncomplicated infectious pustular vulvovaginitis. It is proposed that that multifactorial aetiology, including synergistic pathogenic action between the organisms isolated, was responsible for this outbreak. Administration of oxytetracycline to affected animals was followed by prompt resolution of symptoms. No further outbreaks have been reported.

Mouse embryos cultured in amniotic fluid.

One thousand mouse embryos were collected over a period of 6 weeks and randomly assigned to either amniotic fluid aspirated during routine amniocentesis from normal 16 weeks pregnant patients or Earle's medium. The embryos were cultured for 72 hours at 37 degrees C in 5% carbon dioxide in air. Osmolarity, pH, partial arterial carbon dioxide pressure and HCO3- were checked before culture and again at the end of culture. Embryos were assessed according to the stage of development, namely degenerate embryos, morulae and blastocysts. The blastocysts were further divided according to their stage of development. There was a significant improvement in blastulation rate to fully expanded blastocysts in the embryos cultured in the amniotic fluid. The amniotic fluid was fairly variable in composition, pH and osmolarity, whereas the Earle's medium was very uniform in composition between trials. It was concluded that amniotic fluid drawn at 16 weeks from normal women is a good culture medium for mice embryos.

Changes in sperm motility, morphology and concentration induced by the swim-up procedure.

Fifteen semen samples from normal donors were washed twice in Earle's medium by centrifugation and allowed to swim up into overlayered Earle's medium. Aliquots of 0.2 ml were drawn before and after the swim-up, for assessment of sperm motility, morphology and concentration. The percentage of sperm showing progressive motility improved by an average of 54.02 +/- 45.97%, and sperm with the normal morphology increased by 63.8 +/- 31.01%. Head and tail abnormalities decreased by 47.17 +/- 21.3% and 89.37 +/- 16.1% respectively. There was no statistical change in acrosomal abnormalities. Sperm concentration was reduced by 89.02 +/- 7.4%. It is concluded that the standard swim-up technique resulted in an overall improvement in sperm quality, but that the technique was not effective in reducing the acrosomal abnormalities.

Influence of the swim-up procedure on acrosome damage induced by freezing human semen.

Twenty frozen semen samples from normal donors were thawed, washed in Earle's medium, and allowed to swim up using standard techniques. The samples were evaluated before and after the swim up for progressive motility and concentration. Morphologically normal sperm were assessed for acrosomal integrity according to the relative percentages of normal, damaged or lost acrosomes. The motility improved by 75.07 +/- 90.38%, the count was reduced by 97.1 +/- 1.9%, but there was no significant change in the frequencies of the different types of acrosome state. It was concluded that although the swim-up technique is successful in reducing most major morphological abnormalities, influence over the frequencies of secondary acrosomal abnormalities could not be demonstrated.

A retrospective study of 40 cases of canine pyometra-metritis treated with prostaglandin F-2 alpha and broad-spectrum antibacterial drugs.

Bitches with pyometra were treated with dinoprost, a prostaglandin F-2 alpha-THAM salt or luprostiol, a synthetic PG analogue and oral broad-spectrum antibacterial drugs. PGF-2 alpha treatment lasted 2-26 days and dosage varied from 26.8 to 258 micrograms/kg. Clinical cure from symptoms was achieved in 33/40 bitches. Of 14 bitches from which the owners opted to breed, 9 eventually produced litters. Long-term complications were anoestrus, recurrence of metritis, failure to conceive and abortion. One or more of these complications were recorded in 9 of 20 bitches for which long term information was available.

Luteolysis in early diestrous beagle bitches.

To induce luteolysis early in diestrus, 15 mated beagles were treated with 250 microg/kg prostaglandin F2alpha (PG), administered subcutaneously twice daily for 4 d, from Day 5 to Day 8 after the onset of cytological diestrus. They were divided into three groups of five bitches each: Group 1 received PG only. Group 2 received PG and 20 microg/kg/day bromocryptine orally. Group 3 received PG and 0.1 mg/kg/day dexamethasone intramuscularly. Fifteen untreated beagles served as controls. In each of the three treatment groups, four of the five bitches (80%) underwent complete luteolysis and did not whelp. The three bitches which did not undergo luteolysis whelped normally. All control bitches whelped normally. It was concluded that luteolysis is possible in bitches early in diestrus and the inclusion of bromocryptine or dexamethasone did not appear to influence luteolysis.

Ultrastructural changes in the acrosome of human sperm during freezing and thawing: a pilot trial.

As a pilot trial, a semen sample was diluted with a glucose-citrate-egg yolk diluent and frozen in 0.5 ml PVC paillettes in liquid nitrogen vapor. Motility and acrosome integrity of the sample were evaluated before and after freezing, and longevity was monitored up to 6 h postthaw. Acrosome integrity was assessed by comparing Spermac-stained thin smears with TEM. Acrosome damage was found to be progressive, and four main types of acrosome state were identified and illustrated.

A pregnant heifer with a congenital incompletely developed vulva.

A 2 year old Friesland heifer was presented in an advanced stage of pregnancy with a vulva showing only 2 small openings of 1,5 and 0,5 cm in diameter in the dorsal and ventral vulvar commissures, respectively. Normal vulvar skin covered a thick band of fibrous tissue between the two openings. From the vestibulum forward, the rest of the genital tract was normal. At calving, the fibrous tissue was resected under epidural anaesthesia, and the calf delivered with ease. The vulva was reconstructed by stitching the vestibular mucosa to the vulvar skin using simple interrupted sutures.

Infertility in a Maltese poodle as a result of a sperm midpiece defect.

A 2 1/2-year-old Maltese poodle was examined for breeding soundness following a series of unsuccessful matings to fertile bitches. He was found to have only 8% normal sperm in his ejaculate when Spermac-stained smears were examined under the light microscope. The defect most frequently encountered involved the midpiece attachment, and the various manifestations of disintegration found in this region are described and illustrated. Transmission and scanning electron microscopy confirmed light microscopical findings.

Preliminary report on the effect of prostaglandin F(2)alpha on the duration of the oestrus interval in beagle bitches.

In an attempt to shorten the oestrus interval of bitches, ten nonpregnant beagles were treated with prostaglandin F (PGF(2)alpha) within eight weeks of oestrus. The dose varied from 60 to 500 mug/kg/day administered over three to six days. Fifteen untreated bitches served as controls. The average oestrus interval of treated bitches was four months, while that of the controls was 6.55 months.

Preliminary report: a pregnancy from frozen centrifuged dog semen.

Semen obtained from a pure-bred Beagle was diluted 1:3 with a Tris (hydroxymethyl) - aminomethane buffered egg yolk diluent, centrifuged at 450 g for 10 minutes and the spermatozoa thus concentrated stored in 8 Cassou mini straws (0,25 ml) in liquid nitrogen. One month after collection, a single semen straw was thawed, rediluted with 4 ml of diluent containing no glycerol and inseminated into a Beagle in oestrus. This was repeated daily for 4 days. The vagina of the bitch was flushed with 4 ml of the diluent immediately prior to insemination on each occasion. The bitch whelped 7 pups 60 days after the first insemination.

Clinical experience with prostaglandin F2 alpha THAM as a luteolytic agent in pregnant and non-pregnant bitches.

Prostaglandin F 2 alpha THAM was administered to 6 bitches at various stages after service at dosages ranging from 30-250 micrograms/kg for 2-6 days. Four bitches treated between Days 21-42 after second service did not produce pups irrespective of dosage and duration of administration. A bitch treated in the first trimester and another in the third trimester did not abort but gave birth to normal litters. Two unserved bitches were also treated in the first 20 days of metoestrus with no drop in plasma progesterone being evident. It would seem that prostaglandin F2 alpha THAM is useful as a luteolytic agent only in the second trimester. Although side effects of mild panting with occasional vomiting and/or defecation were sometimes noted at all dosage levels, they were milder at the recommended lower dosage of 30 micrograms/kg administered twice daily subcutaneously for 4 days.

A modified Shorr's stain: a practical rapid stain for canine vaginal cytology.

A simple modification of Shorr's stain for vaginal cytology is described. The technique involved 8 steps and takes less than 10 minutes from time of making the smears until time of examination. The method is considered to be quick, easy, accurate and practical for use by the small animal practitioner.