BACKGROUND: Canine 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. RESULTS: Total 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. CONCLUSIONS: This 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.

A case of non-syndromic, complete syndactyly involving all four limbs is described in a three-month-old male crossbreed dog for the first time. Syndactyly is a rare condition in most animal species, in dogs it has been infrequently reported. Findings of clinical, radiographic and cytogenetic analyses are described and demonstrate probably for the first time that numerical and structural chromosome aberrations are not involved in the pathogenesis of this case of syndactyly

Background: Surgical 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. Results: Testicles 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. Conclusions: A 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.

Due to the increased attention that pet-owners devote to their animals and to the improved veterinary care, investigations regarding methods to early detect prostatic disorders that might affect canine life quality have been performed. Canine prostate specific esterase (CPSE) concentration was reported to be higher in dogs suffering from prostatic diseases. This study aimed to estimate the CPSE threshold as a biomarker to early identify prostatic diseases in asymptomatic dogs. The ultrasonographic examination of the prostate was performed in 19 dogs (6–40 kg; 1–5 years) with no symptoms of prostatic diseases. Dogs were grouped according to the presence (Group A) or absence (Group B) of prostatic disorders at the ultrasound (altered appearance, the presence of cysts or irregular borders). For each dog, a venous blood sample was collected to measure serum CPSE and the ratio between calculated and normal expected prostatic volume was assessed for each dog. The CPSE data were statistically analysed (t test, p < .05), and the CPSE threshold in blood serum between groups was calculated by ROC. In 11 dogs, ultrasonography showed signs of prostatic abnormalities (Group A, 2–5 years), while no signs were detected in eight dogs (Group B, 1–3 years). The calculated/estimated volume ratio resulted greater than 1.5 in Group A dogs. The CPSE was statistically different between groups (p < .0001): higher in Group A (mean = 184.9, SD = 126 ng/ml) than in Group B (38.9 ± 22.1 ng/ml). The cut-off CPSE threshold was 52.3 ng/ml (ROC, AUC = 0.974, SE 95.6%, SP 89.2%). This study suggests that CPSE serum concentration higher than 50 ng/ml in asymptomatic dogs is associated with ultrasonographic alterations and increased the prostatic size (volume by 1.5 times greater than the normal size). As the onset of prostatic disorders often remains asymptomatic, the rapid assessment of CPSE could be suitable for selecting preventively those animals that would require further accurate evaluation.

An affordable and effective non-surgical technique for achieving male dog sterility is needed to solve the problem of overpopulation. The efficacy of 20% calcium chloride in pure alcohol solution, injected into the testicular parenchyma, as a method for chemical castration, was evaluated. Twenty-one dogs of mixed breed, 4.7 ± 1.23 years old, 20 ± 5.84 kg of body weight, with good clinical conditions and normal reproductive parameters, were lightly sedated and injected into the dorsocranial portion of both testes with a solution of 20% calcium chloride dihyrdate in ethanol (95%). The dose injected corresponds with the testicular width (19–22 mm receive 0.8 ml; 23 and above 1 ml). Semen evaluation was performed by CASA (Computer Assisted Sperm Analysis) system at day 30–60–90. The animals in the control group received a single bilateral intratesticular injection of 1 ml sterile saline solution (testicular width 23 mm and above). Forty-eight hours after the injection, dogs showed very light discomfort at palpation and testicular tumefaction, which regressed within 3 days. At day 30, testicular ultrasonography revealed bilateral more dense nodular lesions; prostatic volume and parenchyma were normal. Semen evaluation showed azoospermia at day 30–60 and 90. The sperm count was decreased significantly (p < 01) in all the CaCl2 treated dogs in comparison to saline solution control animals. At day 90 testicles were shrunk at palpation. An intratesticular injection of 20% calcium chloride in pure alcohol solution, as a method for chemical castration, was effective and economical for the sterilization of male dogs. It is free from pain and chronic stress and will contribute to a simple alternative method to surgical castration. The dogs of this study are under evaluation to study this solution long term effect (1 year).

The buffalo has a seasonal reproduction activity with mating and non-mating periods occurring from late autumn to winter and from late spring to beginning of autumn, respectively. Sperm glycocalyx plays an important role in reproduction as it is the first interface between sperm and environment. Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. We adopted a novel procedure in which spermatozoa was spotted on microarray slides, incubated with a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin, and then signal intensity was detected using a microarray scanner. Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods.

BACKGROUND. 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. METHODS. PEMF (5 min, twice a day for 3 weeks) was performed on 20 dogs affected by BPH. Prostatic volume, Doppler assessment by ultrasonography, libido, semen quality, testosterone levels, and seminal plasma volume, composition and pH were evaluated before and after treatment. RESULTS. The 3 weeks of PEMF produced a significant reduction in prostatic volume (average 57%) without any interference with semen quality, testosterone levels or libido. Doppler parameters showed a reduction of peripheral resistances and a progressive reduction throughout the trial of the systolic peak velocity, end-diastolic velocity, mean velocity, mean, and peak gradient of the blood flow in the dorsal branch of the prostatic artery. The pulsatility index and the resistance index did not vary significantly over time. CONCLUSIONS. The efficacy of PEMF on BPH in dogs, with no side effects, suggests the suitability of this treatment in humans and supports the hypothesis that impairment of blood supply to the lower urinary tract may be a causative factor in the development of BPH. Prostate 74:1132–1141, 2014. # 2014 The Authors. The Prostate published by Wiley Periodicals, Inc. This is an open access article under the terms of the Creative Commons Attribution- NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

Abstract The aim of the present study was to assess the effects feeding level on body weight changes and semen parameters in adult Sardinian rams reared under intensive conditions in a semi-arid area of southern Italy. During an experimental period of 90 days, 24 healthy Sardinian rams were divided into three equal groups that differed in their feeding level, in terms of concentrate amount. The controlconcentrate (CC; n=8) group received 1.0 times their maintenance requirements, the medium-concentrate (MC; n=8) group received a diet that supplied 1.2 times their maintenance requirements, and high-concentrate (HC, n=8) group received a diet that supplied 1.5 times their maintenance requirements. Mixed vetch–oat hay was offered ad libitum to ram groups and water and mineral licks were freely available. Body weight and feed intake was recorded weekly, and semen characteristics were determined every 2 weeks. Dietary treatment affected final body weight (P<0.01) as feeding level increased. Total dry matter and protein intake changed significantly (P<0.05 and P<0.01, respectively) among experimental groups. Semen volume and concentration were positively influenced by feeding level for HC group, whereas no differences were observed in sperm viability and scrotal circumference of rams. It was concluded that dietary level with higher concentrate supplementation resulted in improved body weight gain, feed intake, sperm production, and semen quality in Sardinian rams.

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.

ABSTRACT - The purpose of this review article is to present a 50-year perspective of research on mechanical method for limiting male dog reproduction by the use of therapeutic ultrasound. Ultrasound's potential as a male contraceptive was first reported by Fahim et al. in 1977 where it was shown that a single application could result in a reversible dramatic loss of germ cells. If the method can be made permanent, a noninvasive method for controlling domestic pet populations could be developed, although standard treatment is not yet identified. More recent studies in 2000s by our research group demonstrated that tree treatment

An ideal contraceptive for male dogs should be 100% efficient, irreversible, inexpensive and with no side effects. Ultrasound was used as a male contraceptive in several species, including dogs. Studies are needed to determine minimal number of treatments, interval between treatments, part of testes that requires treatment, frequency or power of ultrasonic wave exposure and many more need to be investigated before its application in practice. A previous study concluded that three treatments of 5 min/treatment per day (2.5 cm2 transducer, 1 MHz, 1.5 W/cm2) are ineffective. In this trial, effects of two testicular ultrasound exposure protocols (differing in ultrasound treatment length and number of applications) on testicular size, consistency and volume, and sperm concentration and motility were evaluated. Twenty dogs were divided in two equal groups- A and B. All subjects were exposed to 1.5 Wcm2 of ultrasound on each testicle using Vetrison Portable ultrasound (Physiomed Elektromedizin AG, Germany; 2.5 cm2 transducer). Dogs in group-A received ultrasound for 5 min on alternate days for one week and dogs in group-B received ultrasound for 15 min inutes twice a day on alternate days. Sperm concentrations and motility evaluations were made before and 25 days after the end of treatments. Length and testicular width were echographically measured to calculate the volume. Semen collected was examined by using an integrated visual optical system for semen analysis for sperm concentration and for percentage of total and progressively motile sperms. All dogs were castrated at day 40 and gonads were collected for histological examination. Data concerning testicular volume were statistically analyzed with ‘Wilcoxon matched pairs signed rank sum’ test (p £ 0.05); semen evaluation was statistically analyzed with ANOVA test (p £ 0.01). After ultrasound treatment, all dogs showed no local or systemic adverse effects, and no pain or skin burns. However, dogs in group A exhibited marked tenderness of testicles at palpation. Group A dogs showed a statistically significant reduction of the volume of both testis (left 9.6 ± 3.7 vs. 5.5 ± 3.6/cm3; right 9.6 ± 2.7 vs. 3.6 ± 1.4/cm3; p £ 0.05) while no reduction in testicular volume was noticed in dogs in group (left: 9.1 ± 1.4 vs. 9.2 ± 1.6/cm3; right: 9.3 ± 1.5 vs. 9.4 ± 1.7/cm3). Before the US treatment, mean volume of ejaculates was 10 ± 3.5 ml, sperm concentration was 300.8 ± 24.8 · 106/ml with an average percentage of total and progressive motile sperms of 88.2 ± 4.5 and 59.3 ± 5.3, respectively. After the ultrasound treatment, a zero sperm count was noticed in group A dogs (p £ 0.01), and no variation in B group. Histology evaluation showed interstitial fibrosis, widespread tubular atrophy and hyalinization of the basement membranes in group-A dogs and no changes were observed in group-B dogs. Our results demonstrated that ultrasound treatment for 5 min on alternate days for one week leads to irreversible testis damage consistent with permanent sterilization, while reducing the number of applicationseven with a longer treatment was ineffective on dog fertility

L’esperienza svolta nella ideazione, gestione e realizzazione del progetto pilota “Tutela e Riproduzione del Riccio di Mare (Paracentrotus lividus) specie innovativa per l’acquacoltura” dalla Algesiro srl di Manfredonia e della Facoltà di Medicina Veterinaria Dipartimento di Benessere Animale dell’Università di Bari è un modello per sviluppare la filiera ittica e la collaborazione tra imprese e mondo della ricerca. Classificato al primo posto della graduatoria nella valutazione di merito, avendo colto il senso del bando ed i suoi obiettivi, il progetto è stato condotto con puntualità ed aggiornamento costante rispetto alle scoperte via via effettuate ed alle tecnologie applicate. L’esperienza si colloca nelle buone pratiche che la Regione Puglia intende adottare e replicare perché il settore delle risorse ittiche possa crescere in maniera sostenibile e nel lungo periodo. L’attenzione posta alla tutela dell’ambiente e delle risorse partendo dalla richiesta di mercato del riccio di mare dimostra che è possibile progettare la gestione della filiera ittica bilanciando gli interventi tenendo conto della pluralità degli interessi e della qualità. Il protocollo sviluppato, primo al mondo, è replicabile in ogni angolo del pianeta e la funzione degli echinodermi, sia in allevamento sia in banchi naturali, contribuisce a tutelare l’ecosistema. Con l’auspicio che esperienze positive come questa possano replicarsi attivando rapporti virtuosi tra imprese, mondo della ricerca, istituzioni e ambiente per la crescita del territorio pugliese

In this study, ultrasonographic examination was performed thrice, 15days apart, on juvenile European sea bass Dicentrarchus labrax, from 330 to 360days 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.

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.