Use of Contrast-Enhanced Ultrasound of the Testes after Non-Surgical Sterilization of Male Dogs with CaCl2 in Alcohol.

Sterilization by intratesticular injection of chemical agents is a non-surgical alternative to neutering male companion animals. We used contrast-enhanced ultrasound (CEUS) to monitor vascular alterations to testes immediately after the intratesticular injection of CaCL2 in alcohol. We evaluated the CEUS features of normal and damaged testes in 20 dogs after the intratesticular injection of CaCl2. The CEUS evaluation was performed at the site of the chemical agent inoculation. In treated testes, qualitative CEUS showed a lower intensity enhancement of the parenchyma than pre-treatment normal testes with a predominantly anechoic pattern and only a few hyperechoic vascular focal spots. Quantitative CEUS showed significantly lower values of time-intensity curve (TIC) parameters, including signal intensity (Peak: 4.72 ± 2.1), regional blood volume (RBV: 134.3 ± 63.7), and regional blood flow (RBF: 4.36 ± 2.18) than normal testes (p < 0.001). Sonographic findings from CEUS showed hypovascularization of the canine testicular parenchyma caused by the hardening agent. This diagnostic technique helps clinicians define testicular vascular alterations achieved by chemical castration more efficiently. Nevertheless, more studies are required to apply this methodology to more subjects with a broader weight range and stray dogs.

Pre- and Probiotics to Increase the Immune Power of Colostrum in Dogs.

Wide differences in Ig concentration in canine colostrum have been reported. Thus, some litters can be at risk of passive immune transfer failure. Present study evaluated if supplementation with MOS, FOS, E. faecium and L. acidophilus along pregnancy increases colostrum quality. Twenty Great Dane bitches were divided into 4 groups. Control group (CG) received standard diet, only. Diet was supplemented with pre- and probiotics in other 3 study groups during: the last (1WG), last 2 (2WG), and last 4 (4WG) weeks of pregnancy, until parturition. Serum samples were collected at estrous (T0), supplementation beginning (T1), and parturition (T2). Colostrum was collected at C-section end. The IgG, IgM, and IgA were assayed on both matrices. In serum, IgG were higher at T0 than at parturition in all study groups and they significantly lowered from T0 to T1 in all groups. In colostrum, IgG and IgM were significantly higher in 4WG, while IgA already increased in 2WG group. Four-week pre- and probiotic supplementation resulted in the best immune properties of colostrum, as by the higher IgG, IgM, and IgA colostrum levels found in 4WG. Further studies would verify the exact mechanisms involved: pre-partum IgG mammary accumulation and B-cells GALT proliferation and mammary transfer. Further trials would verify whether these beneficial effects of pre- and probiotics on colostrum also lead to improved clinical conditions and immunological functions of newborns and puppies.

Ejaculation Effect on Canine Prostatic Specific Esterase Serum Concentration.

Canine prostatic diseases are usually asymptomatic in their onset and often identified in advanced stages. Canine prostatic specific esterase (CPSE) represents an early serum marker for prostatic diseases, also in asymptomatic dogs. The present study aimed to identify the effects of ejaculation on serum CPSE. Twenty adult intact male dogs were enrolled. Blood samples were collected to measure CPSE concentrations before (T0), immediately after (T1), and 24 h post (T2) ejaculation. Data were compared within and between groups by ANOVA (p < 0.05). Dogs were divided in two equal groups: A (healthy: CPSE ≤ 52.3 ng/mL at T0) and B (suspected for prostatic disorders: CPSE > 52.3 ng/mL or diagnosed with symptoms of prostatic diseases: CPSE > 90 ng/mL). CPSE was shown to be statistically higher in group B than A at any time point. In both groups, CPSE showed a significant increase at T1, and no significant differences between T0 and T2. This study demonstrates a definite effect of ejaculation on CPSE concentration. Twenty-four hours post-ejaculation, CPSE returns to basal values. Such physiological effects of ejaculation should be considered when planning analyses of CPSE concentrations, i.e., by respecting a proper sexual rest.

Effects of intratesticular vs intraepididymal calcium chloride sterilant on testicular morphology and fertility in dogs.

BACKGROUND: Both stray and free-roaming owned dogs contribute to the serious global dog overpopulation problem. Many dog owners are unwilling to have their pet castrated for various reasons, including a reluctance to have their dog's behavior changed. A non-surgical method of sterilizing both stray and owned dogs would help to prevent unwanted litters. Previous studies have shown that intratesticular injection of calcium chloride dihydrate (CaCl2) in alcohol is a promising and cost-effective alternative to surgery for stray dogs, with testosterone significantly decreased and sexual activity eliminated. The aim of this study was to compare the use of a solution of 20% CaCl2 in 95% ethanol injected into the testicles or into the head of the epididymis. METHODS: A total of 148 dogs divided into 4 groups (2 experimental and 2 control) were respectively injected with CaCl2 or saline solution into the testicle or epididymal head (ultrasound-guided). The animals were examined at 0, 3, 6, and 9 months for sperm quality, concentration of testosterone in serum, and side effects; at 0 and 5 months with contrast-enhanced ultrasound (CEUS) to enhance the morphological aspects/alteration of the testicular parenchyma or epididymis; and at 9 months when all were castrated for histological examination. RESULTS: All dogs treated with CaCl2 became sterile with azoospermia achieved over the 9-month study. The concentration of testosterone in serum significantly decreased following intratesticular treatment with CaCl2. No adverse effects were noted. CONCLUSIONS: A single, bilateral intratesticular injection of 20% CaCl2 in 95% ethanol was confirmed to be a reliable method for induction of sterilization in male dogs. The approach showed long-term efficacy and may reduce sexual behavior, with the additional benefits of low-cost and ease of use, making this nonsurgical method appropriate for use in stray dogs. Sterility was also achieved if injected in the head of the epididymis but no significant decrease in serum concentration of testosterone occurred. Moreover, performing the intraepididymal injection into the epididymal head was as time consuming as orchiectomy. This approach may be optimal for use in owned dogs where anatomical integrity and testosterone maintenance is preferred by the owner.

The Effect of Dietary Supplementation of Vitamin E, Selenium, Zinc, Folic Acid, and N-3 Polyunsaturated Fatty Acids on Sperm Motility and Membrane Properties in Dogs.

Sub-fertility represents a common challenge in canine reproduction. Different protocols, supplementing daily given quantities of micronutrients, were investigated to improve poor sperm concentration and/or function, which represent breeding major constraining factors in the canine species. Little information is available for dogs concerning the effect of a daily supplementation with a complex of vitamin E, zinc, selenium, folic acid, and n-3 polyunsaturated fatty acids (PUFA) on semen quality. Thus, the present study investigated this effect on semen motility and sperm membrane properties. Serial semen analyses from fourteen healthy normospermic dogs, fed with the same commercial diet, were performed on Days 0 (T0), 30 (T30), 60 (T60), and 90 (T90). Seven dogs were randomly included in the treatment (T) group, receiving a supplementation of vitamin E, zinc, selenium, folic acid, n-3 PUFA; and seven other subjects composed the control (C) group. Total Sperm Count (TSC), Computer-Assisted Sperm Analysis (CASA) indexes, mortality, and functional membrane integrity were assessed. The ANOVA compared results between groups and sampling times (p < 0.05). From T60, the supplementation significantly improved TSC, progressive motility, functional membrane integrity, and decreased mortality. Present results lead us to consider ameliorative effects of a two-month healthy diet supplementation on canine spermatozoa. The positive effects of the described balanced integration of micronutrients on sperm motility and prevention of oxidative stress should be considered, especially when decreased seminal parameters may result from inadequate intake, reduced absorption, increased losses or demand, or to attenuate the impact of age.

Ejaculation effect on blood testosterone and prostatic pulsed-wave Doppler ultrasound in dogs.

Local vessels ultrasonography evaluates prostatic physio-pathologic states. Testosterone promotes tissue and vascular growth. Knowing variables on prostatic vasculature is crucial to correctly apply Pulsed-Wave exam. The study aims to assess how ejaculation and blood testosterone affect Pulsed-Wave indexes. Serial blood testosterone dosages and Pulsed-Wave exams were performed in 20 dogs, immediately before (T0) and after (T1) ejaculation and 6 hr later (T2). Arteria prostatica cranialis, Arteriola capsularis, Arteriola trabecularis and Arteriola parenchimalis were evaluated and mean Pulsatility and Resistivity Index, Systolic-Peak, End-Diastolic and Mean Velocity calculated. Data were grouped by time and vessel (ANOVA, p ≤ 0.05). At T1, Resistivity Index significantly lowered in A. prostatica cranialis, A. trabecularis and A. parenchimalis but grew in A. capsularis; Pulsatility Index had the same pattern, but not significant in A. parenchimalis; Systolic Peak Velocity, End-Diastolic Velocity, Mean Velocity significantly rose in A. capsularis and A. trabecularis. No indexes differed at T0 and T2. Testosterone did not differ at T0 (10.93 ± 7.05 ng/ml), T1 (12.71 ± 7.29) and T2 (10.54 ± 6.63). Results stated the risen prostatic vascular flow postejaculation, affecting Pulsed-Wave. Due to semi-rigid capsule, impairing vasodilation of other vessels, only A. capsularis indexes increased. Intimal cushions of A. prostatica cranialis kept velocities fixed; A. capsularis and A. trabecularis lack of intimal cushions, thus velocities grew. In A. parenchimalis, precapillary sphincters opening allows increased flow redistribution in vasodilated parenchymal bed, keeping velocities fixed. As testosterone, not affected by ejaculation, did not peak, vascular changes are not due to testosterone itself. These physiological effects of ejaculation suggest proper sexual rest before Pulsed-Wave exam planned to explore suspected prostatic neovascularization.

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

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.

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.

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.

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.

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.