Giulio Aiudi

Cumulus expansion, nuclear maturation and connexin 43, cyclooxygenase-2 and FSH receptor mRNA expression in equine cumulus-oocyte complexes cultured in vitro in the presence of FSH and precursors for hyaluronic acid synthesis

The aim of this study was to investigate cumulus expansion, nuclear maturation and expression of connexin 43, cyclooxygenase-2 and FSH receptor transcripts in equine cumuli oophori during in vivo and in vitro maturation in the presence of equine FSH (eFSH) and precursors for hyaluronic acid synthesis. Equine cumulus-oocyte complexes (COC) were cultured in a control defined medium supplemented with eFSH (0 to 5 micrograms/ml), Fetal Calf Serum (FCS), precursors for hyaluronic acid synthesis or glutamine according to the experiments. After in vitro maturation, the cumulus expansion rate was increased with 1 microgram/ml eFSH, and was the highest with 20% FCS. It was not influenced by precursors for hyaluronic acid synthesis or glutamine. The expression of transcripts related to cumulus expansion was analyzed in equine cumulus cells before maturation, and after in vivo and in vitro maturation, by using reverse transcription-polymerase chain reaction (RT-PCR) with specific primers. Connexin 43, cyclooxygenase-2 (COX-2) and FSH receptor (FSHr) mRNA were detected in equine cumulus cells before and after maturation. Their level did not vary during in vivo or in vitro maturation and was influenced neither by FSH nor by precursors for hyaluronic acid synthesis. Results indicate that previously reported regulation of connexin 43 and COX-2 proteins during equine COC maturation may involve post-transcriptional mechanisms.

Relationship Between Motility and Mitochondrial Functional Status in Canine Spermatozoa

Inner mitochondrial membrane potential (IMM) is considered a sensitive indicator for the energetic status and motility of spermatozoa. The relationship between sperm motility parameters evaluated by Computer Assisted Sperm motility Analyzer and plasma membrane integrity and IMM assessed by triple staining (PI/SYBR-14 and JC-1) was evaluated in 10 dogs of unknown fertility. Sperm motility showed large variations ranging from 10% to 98%. Proportion of viable sperm cells and of spermatozoa with high IMM ranged from 74% to 99% and from 53% to 87%, respectively. The presence of a high IMM assessed by JC-1 was more strongly correlated to sperm viability (r = 1) than to sperm motility (r = 0.778). Our results indicate that JC-1 is suitable for detection of IMM changes in canine spermatozoa, but it should always be associated with an objective motility analysis to avoid incorrect evaluation of potential sperm fertility. Ejaculates with a low motility rate showed an unexpectedly high proportion of sperm with high IMM, suggesting that mitochondrial respiration could not be sufficient to support sperm motility, although it may be important for sperm survival in the female genital tract.

Alcohol diluent provides the optimal formulation for calcium chloride non-surgical sterilization in dogs

BackgroundSurgical castration is widely used to sterilize male dogs, but has significant impacts on time to perform the operation, recovery of the animals as well as cost, which can limit population control programs. Previous research has shown intratesticular injection of calcium chloride dihydrate (CaCl2) in saline to be a promising alternative to surgery. However, long-term azoospermia was not maintained at dosages low enough to avoid side effects. In the search for an optimized formulation, the current investigation is the first study on long-term sterilization effects of intratesticular injection of CaCl2 in either lidocaine solution or alcohol in dogs. CaCl2 at 20% concentration in lidocaine solution or alcohol was administered via intratesticular injection to groups of 21 dogs each. The treated animals were examined at 2, 6, and 12 months for sperm production, blood levels of testosterone, and side effects; at time zero and 12 months for testicular size and semen volume. The experimentally treated animals were compared to a control group receiving saline injection only.ResultsTesticles of dogs treated with CaCl2 in either diluent significantly decreased in size. After administration of CaCl2 in lidocaine solution, sterility was achieved for at least 12 months in 75% of treated dogs. However, optimal long-term contraceptive effectiveness was achieved with CaCl2 in alcohol, which resulted in azoospermia over the 12-month study period. Testosterone levels significantly decreased following treatment with CaCl2, and sexual activity disappeared. Although testosterone returned to baseline levels by 12 months for the group treated with CaCl2 in lidocaine, dogs injected with CaCl2 in alcohol had a 63.6% drop in testosterone level, which remained at the low end of physiological range throughout the study. No adverse effects were noted.ConclusionsA single, bilateral intratesticular injection of 20% CaCl2 in 95% ethanol was a reliable method for induction of sterilization in 18-28 kg male dogs in this study. The approach showed long-term efficacy and reduced sexual behavior. This chemical method of sterilization might provide an effective, efficient alternative to surgical castration that can have positive impacts on dog welfare.

Use of laparoscopy for the evaluation of the reproductive status of tench (Tinca tinca).

The aim of the study was to evaluate the use of laparoscopy for the assessment of the reproductive tract and stage of gonadal development of the tench (Tinca tinca). Laparoscopy was carried out from February to March 2009, on 30 anesthetized tenchs, 15 males and 15 females, of approximately 250 g bodyweight and 30 cm length, using a 1.9-mm rigid cystoscope. Male and female gonads, attached to the swim bladder, were visualized. Testes were orange with a striped appearance. Mature ovaries were cluster-shaped and whitish. Histological samples were collected, from the reproductive organs of both the sexes, with a forceps and showed the presence of gametes at different stages of maturity. The application of this minimally invasive diagnostic imaging technique could clinically be used as a powerful tool to evaluate the gender and to establish the reproductive status of tench and other fish.

A dose-finding, long-term study on the use of calcium chloride in saline solution as a method of nonsurgical sterilization in dogs: evaluation of the most effective concentration with the lowest risk

BackgroundCanine overpopulation is a global issue with serious health and welfare implications. Nonsurgical methods of sterilization could yield positive impacts on this problem, but no long-term data on such methods are available. The objective of the current investigation was to determine the effects of intratesticular injections of calcium chloride dihydrate (CaCl2) in saline in dogs over a one year period. Five concentrations (0%, 10%, 20%, 30%, 60%) of CaCl2 in saline were administered via intratesticular injection to groups of 10 dogs each. Total sperm count and motility, blood levels of testosterone, and side effects were examined at 0, 2, 6, and 12 months post-injection (PI). Testicular size and semen volume were examined at 0 and 12 months PI.ResultsTotal sperm count, semen volume and testosterone showed significant dose-dependent decreases upon treatment with 10%-60% CaCl2 compared with either the control group (0% CaCl2) or baseline for each treatment group. Azoospermia was achieved for at least 12 months PI in 60% and 80% of treated dogs after administration of a 10% and 20% CaCl2, respectively. Treatment with 30% or 60% CaCl2 resulted in azoospermia in 100% of dogs, but more side effects were observed, while no side effects were noticed at lower doses. For each treatment group, testosterone levels had decreased an average of 35%-70% at 6 months following treatment. However, testosterone levels rebounded by the 12-month time point in all groups except the highest dosage group (60% CaCl2), which remained at the low end of physiological range throughout the study. Sperm motility dropped to zero or near zero in all dogs treated with CaCl2. Testicular size was significantly smaller at 12 months PI for all groups when compared to baseline.ConclusionsThis first long-term study confirms reports of the efficacy of CaCl2 sterilization. However, at dosages free of adverse events, calcium chloride in saline may not provide permanent sterilization as previously believed. Future work should explore optimized solvents to increase the permanence of the well-tolerated 20% formulation.

Effect of Pulsed Electromagnetic Field Therapy on Prostate Volume and Vascularity in the Treatment of Benign Prostatic Hyperplasia: A Pilot Study in a Canine Model

Benign prostatic hyperplasia (BPH) is a result of urogenital aging. Recent studies suggest that an age‐related impairment of the blood supply to the lower urinary tract plays a role in the development of BPH and thus may be a contributing factor in the pathogenesis of BPH. The canine prostate is a model for understanding abnormal growth of the human prostate gland. We studied the efficacy of pulsed electromagnetic field therapy (PEMF) in dogs to modify prostate blood flow and evaluated its effect on BPH.

Ultrasound as a Mechanical Method for Male Dog Contraception

Therapeutic ultrasound (1.5 W/cm(2)) was investigated for its effect upon testicular size and consistency and semen quality in five dogs. Dogs were treated every other day with a total of three treatments and were examined up to 2 weeks after treatment. There was some tenderness after treatment and a significant decrease in testicular size and semen quality. This method may provide a suitable contraception technique in dogs and requires further investigation.

Mu opioid receptor in spermatozoa, eggs and larvae of gilthead sea bream ( Sparus Aurata ) and its involvement in stress related to aquaculture

Abstract In aquaculture, unfavourable conditions experienced during early development may have strong downstream effects on the adult phenotype and fitness. Sensitivity to stress, leading to disease, reduced growth and mortality, is higher in larvae than in adult fish. In this study, conducted on sea bream (Sparus aurata), we evidenced the presence of the mu opioid receptor in gametes and larvae at different developmental stages. Moreover, we evaluated the possibility of reducing the effects of artificially produced stress, altering temperature, salinity and pH, by naloxone (an opioid antagonist) and calcium. Results evidenced that mu opioid receptor is present in larvae and in gametes of both sexes and that, during larval growth, its expression level changes accordingly; furthermore, naloxone/calcium association is efficacious in increasing the survival period of treated larvae compared to controls. We conclude that in sea bream rearing, the use of naloxone/calcium against stress can improve fish farming techniques by reducing larval mortality and consequently increasing productivity.

Induction of sexual activity in male and female farmed ostriches (Struthio camelus) with GnRH implant

Abstract The annual photoperiodic changes are the most important factor controlling the reproductive activity in birds. A single subcutaneous implant of a slow-release GnRH analogue induced the seasonal reproductive activity in both sex of farmed ostriches. Significant increase in annual egg production and clutch number compared with control group were observed.

Ultrasound measurement of reproductive organs in juvenile European sea bass Dicentrarchus labrax

In this study, ultrasonographic examination was performed thrice, 15 days apart, on juvenile European sea bass Dicentrarchus labrax, from 330 to 360 days of age, to assess the size and the morphology of male and female. Results have proved this method as a suitable and non-invasive procedure to assess sexual differentiation.