Effect of deslorelin acetate treatment in oocyte recovery and in vitro embryo production in domestic cats.

Objectives The present study investigated the effect of contraceptive treatment with deslorelin acetate on in vitro embryo production and oocyte recovery in domestic queens. Methods Twenty-one mature domestic cats were used. Eleven queens (treated group) and one tom were kept in an experimental cattery, and 10 queens were privately owned (control group). When in interestrus or diestrus (day 0) a deslorelin acetate implant (Suprelorin, 4.7 mg/animal) was inserted into the subcutaneous tissue of the interscapular region in all queens in the treated group. After 6 months of treatment, all animals were ovariohysterectomized, and the ovaries were used for in vitro embryo production. Percentage of cleavage was determined 18 h after oocyte insemination and blastocyst formation was assessed on the eighth day of culture. The rate of cumulus-oocyte complexes (COCs) recovery was analyzed by an unpaired t-test. The cleavage and blastocyst rates were expressed as percentages and analyzed by Fisher's exact test. All analyses were performed using GraphPad Prism v5.0, with P <0.05 set as the level of significance. Results In the treated group, we recovered 8.3 ± 1.15 grade I COCs per queen; the cleavage rate was 60% and the blastocyst rate was 36%. In the control group, we recovered 18.4 ± 3.21 grade I COCs per queen; the cleavage rate was 55.97% and the blastocyst rate was 34%. Forty percent of treated females did not produce any blastocysts. In the treated group, we observed a significant decrease in COC recovery. Although there was no significant difference in cleavage and blastocyst rates between groups, 40% of treated females did not produce any blastocysts. Conclusions Recovery of grade I COCs is negatively affected by deslorelin treatment in domestic cats. Regarding embryo production, new studies are still necessary to evaluate the success of this technique owing to the individual effect caused by deslorelin acetate.

Time course of the meiotic arrest in sheep cumulus-oocyte complexes treated with roscovitine.

Temporary meiosis arrest with cyclin-dependent kinases inhibitors has been proposed in order to improve the quality of in vitro matured oocytes. In sheep, however, this phenomenon has been rarely investigated. Therefore, the present study aimed to evaluate the effect of different incubation times with roscovitine on nuclear maturation and cumulus cell expansion of sheep cumulus-oocyte complexes (COCs). For this, COCs were cultured for 0, 6, 12 or 20 h in basic maturation medium (Control) containing 75 µM roscovitine (Rosco). After, they were in vitro matured (IVM) for 18 h in the presence of luteinizing hormone (LH) and follicle-stimulating hormone (FSH). At the end of each treatment, cumulus cell expansion and nuclear maturation were assessed under a stereomicroscope and by Hoechst 33342 staining, respectively. In the Control and Rosco groups, the absence of cumulus cell expansion prevailed at 0, 6, 12 and 20 h. After IVM for 18 h, total cumulus cell expansion in the Rosco treatments was dependent on the exposure time to roscovitine. A significantly high percentage of oocytes treated with roscovitine for 6 h (87%), 12 h or 20 h (65%) were arrested at the germinal vesicle (GV) stage. In contrast, 23% GVBD, 54% metaphase I (MI) and 61% MII oocytes were observed in the Control groups at 6, 12 and 20 h, respectively. In all treatments, a significant percentage of oocytes reached MII after IVM for 18 h. Therefore, roscovitine reversibly arrested the meiosis of sheep oocytes during different culture times with the maximal efficiency of meiotic inhibition reached at 6 h. In addition, reversibility of its inhibitory action on cumulus cells was exposure-time dependent.

Semen quality, testicular B-mode and Doppler ultrasound, and serum testosterone concentrations in dogs with established infertility.

Retrospective examination of breeding records enabled the identification of 10 dogs of normal fertility and 10 dogs with established infertility of at least 12 months of duration. Comparisons of testicular palpation, semen evaluation, testicular ultrasound examination, Doppler ultrasound measurement of testicular artery blood flow, and measurement of serum testosterone concentration were made between the two groups over weekly examinations performed on three occasions. There were no differences in testicular volume (cm(3)) between the two groups (fertile right testis = 10.77 ± 1.66; fertile left testis = 12.17 ± 2.22); (infertile right testis = 10.25 ± 3.33; infertile left testis = 11.37 ± 3.30), although the infertile dogs all had subjectively softer testes compared with the fertile dogs. Infertile dogs were either azoospermic or when they ejaculated, they had lower sperm concentration, sperm motility, and percentage of morphologically normal spermatozoa than fertile dogs. Furthermore, infertile dogs had reduced sperm membrane integrity measured via the hypoosmotic swelling test. Infertile dogs had significantly lower basal serum testosterone concentrations (1.40 ± 0.62 ng/mL) than fertile dogs (1.81 ± 0.87 ng/mL; P < 0.05). There were subjective differences in testicular echogenicity in some of the infertile dogs, and important differences in testicular artery blood flow with lower peak systolic and end-diastolic velocities measured in the distal supratesticular artery, marginal testicular artery, and intratesticular artery of infertile dogs (P < 0.05). Notably, resistance index and pulsatility index did not differ between infertile and fertile dogs. These findings report important differences between infertile and fertile dogs which may be detected within an expanded breeding soundness examination.

Luteal changes after treatment with sub-luteolytic doses of prostaglandin (cloprostenol sodium) in cattle.

This study characterizes the physiological and morphological changes related to partial luteolysis in bovine corpus luteum (CL) after challenges with sub-doses of cloprostenol sodium on Day 6 (D6) of the estrous cycle. Cows (n = 12/treatment) were treated as follows: Control (2 mL, saline, i.m.); 2XPGF (two treatments i.m. 500 µg of cloprostenol sodium 2 h apart) and 1/6PGF (83.3 µg of cloprostenol sodium, i.m., once). Plasma progesterone (P4) concentration, CL volume and blood flow were measured immediately before the treatments, then every 8 h (h) for 48 h. In the Control, P4 concentrations were higher at 48 h than at 0 h. P4 decreased 8h after 2XPGF treatment (P < 0.05), and remained low until the end of the trial. P4 decreased in 1/6PGF between 8 and 16 h (P < 0.05), then began to rebound at 24 h. Luteal volume was higher in Controls at 48 h than at 0 h. Under 1/6PGF, luteal volume decreased at 24 h (P < 0.05) and began to rebound at 32 h. Luteal volume and blood flow were reduced starting at 24 and 32 h, respectively, after 2XPGF treatment (P < 0.05). In this study, we were able to describe the partial luteolysis phenomenon, induced by a treatment of a D6CL with cloprostenol sub-dose.