Clinical Efficacy of the GnRH Agonist (Deslorelin) in Dogs Affected by Benign Prostatic Hyperplasia and Evaluation of Prostatic Blood Flow by Doppler Ultrasound

In six German Shepherds dogs, GnRH agonist implants (Deslorelin) were inserted subcutaneously one month after histological confirmation of benign prostatic hyperplasia (BPH). Prostatic volume (PV), characteristics of ejaculate, serum testosterone concentrations and Doppler parameters of prostatic and subcapsular arteries were detected at different time intervals, for 6 month. The prostatic volume showed a significantly reduction starting at day 37. The decrease in sperm concentration, motility and increase in morphological abnormal sperm were observed from day 22 to day 37, when it was no longer possible to obtain the ejaculate. The values of peak systolic velocity and end‐diastolic velocity in prostatic and subcapsular arteries showed from day 11 a gradual decrease, significant at day 22 until day 37 and reaching the lowest values at day 52 until the end of observation. The power Doppler pixel intensity of both arteries showed a gradual decrease from day 5 until day 52. In particular, a significant decrease was observed for both arteries from day 11. Testosterone serum concentration decreased to undetectable levels by day 11 until the end of the observations. All these Doppler parameters and testosterone values were positively correlated with the prostatic volume. Furthermore, testosterone values were positively correlated with peak systolic velocity, end diastolic velocity and pixel numbers. The use of implants containing GnRH analogues, even in asymptomatic subjects, is effective for the control of BPH and the application of Doppler exam of prostatic blood flow represent an non‐invasive tool for monitoring the response of medical treatment.     

Effects of an extract of Serenoa repens on dogs with hyperplasia of the prostate gland

OBJECTIVE: To determine effects of an extract of Serenoa repens on dogs with prostatic hyperplasia. ANIMALS: 20 mature male dogs with benign prostatic hyperplasia. PROCEDURE: Dogs were assigned to 3 comparable groups on the basis of prostatic volume per kg of body weight and degree of prostatic hyperplasia determined histologically. Dogs in 2 groups were treated for 91 days (8 received 500 mg, PO, q 8 h [1,500 mg/d], and 6 received 100 mg, PO, q 8 h [300 mg/d]). The control group of 6 dogs did not receive medication. Effects of treatment on prostatic volume, prostatic weight, prostatic histologic characteristics, radiographic and ultrasonographic assessment of prostatic size, results of CBC, serum biochemical analyses, and urinalysis, serum testosterone concentration, and semen characteristics were determined. At the termination of the study, all dogs were euthanatized, and necropsies were performed. Investigators conducting tests and interpreting results were not aware of treatment group of each dog. RESULTS: Treatment did not affect prostatic weight, prostatic volume, or prostatic histologic scores, libido, semen characteristics, radiographs of the caudal portion of the abdomen, prostatic ultrasonographs, or serum testosterone concentrations. Results of CBC, serum biochemical analyses or urinalysis, and body weights did not change during treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment with an extract of S repens for 91 days did not significantly affect the prostate gland of dogs. Adverse effects were not evident. Although products containing extracts of S repens are widely advertised for men with prostatic hyperplasia, beneficial or harmful effects of this plant extract were not found in dogs with prostatic hyperplasia.     

Evaluation of Efficacy and Safety of Zinc Gluconate Associated with Dimethyl Sulphoxide for Sexually Mature Canine Males Chemical Neutering

The aim of this research was to evaluate the efficacy of zinc gluconate associated with dimethyl sulphoxide (DMSO) for chemical neutering in canine males. Fifteen sexually mature male dogs were divided in two groups, named control and treated. An injection was administered to both testicles, at a concentration of 26.2 mg zinc gluconate per ml and 0.5% DMSO in the treated group (11 dogs). The control group was given injections of saline solution (four dogs). Clinical examination and blood collection for a haemogram were done both before and after drug injection. There were 12 spermograms performed to analyse sperm motility, sperm vigour, ejaculate volume, testicle size, pathology and sperm concentrations. Libido was also measured. An ultrasound examination and histopathology were performed at the end of the experiment. Dogs’ libido after chemical injection was reduced by over 50%. The spermogram analysis showed final mean results of 14.54% for sperm motility, 0.72 of sperm vigour and 37 150 per million spermatozoa per millilitre, values considered below the necessary levels at which fertilization can occur. Ultrasound and histopathology analyses of testicles for the treated group revealed more intense injuries when compared with the control group, with compromised testicular parenchyma and a decrease of germ cell number leading to total atrophy, indicating that the treatment reduced the fertilizing potential of male dogs, promoting a possible subfertile status.     

Effects of finasteride on size of the prostate gland and semen quality in dogs with benign prostatic hypertrophy

OBJECTIVE: To determine the effect of the 5alpha-reductase inhibitor finasteride on prostatic diameter and volume, semen quality, and serum dihydrotestosterone (DHT) and testosterone concentrations in dogs with spontaneous benign prostatic hypertrophy (BPH). DESIGN: Double-blind placebo-controlled trial. ANIMALS: 9 dogs with BPH. PROCEDURE: Five dogs were treated with finasteride for 16 weeks (0.1 to 0.5 mg/kg [0.05 to 0.23 mg/lb] of body weight, PO, q 24 h); the other 4 received a placebo. Prostatic diameter, measured radiographically, prostatic volume, measured ultrasonographically, semen quality, and serum DHT and testosterone concentrations were evaluated before and during treatment. After receiving the placebo for 16 weeks, the 4 control dogs were treated with finasteride for 16 weeks, and evaluations were repeated. RESULTS: Finasteride significantly decreased prostatic diameter (mean percentage decrease, 20%), prostatic volume (mean percentage decrease, 43%), and serum DHT concentration (mean percentage decrease, 58%). Finasteride decreased semen volume but did not adversely effect semen quality or serum testosterone concentration. No adverse effects were reported by owners of dogs in the study. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that finasteride can be used to reduce prostatic size in dogs with BPH without adversely affecting semen quality or serum testosterone concentration.     

RELATIONSHIP BETWEEN PROSTATOMEGALY, PROSTATIC MINERALIZATION, AND CYTOLOGIC DIAGNOSIS

Canine prostatic disease is commonly evaluated with abdominal ultrasound and radiographs. Mineralization of the prostate is often reported, but the clinical relevance of this finding is currently not known. The purpose of this study was to characterize the relationship between ultrasonographic and radiographic prostate mineralization and the final diagnosis. Medical records of 55 dogs with evidence of prostatomegaly or prostatic mineralization and a cytologic diagnosis were evaluated. Radiographs and ultrasound images were assessed for caudal retroperitoneal lymphadenopathy, vertebral lesions, or other signs of metastasis, and mineralization was assessed semiquantitatively. Twenty-two of 55 (40%) dogs had prostatic neoplasia. Regarding neoplasia, mineralization in neutered dogs had a positive predictive value (PPV) of 100%, a negative predictive value (NPV) of 50%, and a sensitivity and specificity of 84% and 100%, respectively. Mineralization in intact dogs had a PPV of 22%, an NPV of 96%, and a sensitivity and specificity of 67% and 77%, respectively. All neutered dogs with prostatomegaly but not prostatic neoplasia had bacterial prostatitis and were castrated within the previous 3 months. Intact dogs with prostatomegaly and mineralization but not neoplasia had paraprostatic cysts ( n =3), benign prostatic hyperplasia ( n =2) or prostatitis ( n =2). Mineralization score was not indicative of neoplasia. In conclusion, neutered dogs with prostatic mineralization were very likely to have prostatic neoplasia. Intact dogs were unlikely to have prostatic neoplasia if no mineralization was found on radiographs or ultrasound.     

Prostatic Neoplasia: In Search of a Treatment

Introduction:  Complete prostatectomy with healing and normal urinary function is reported for some prostatic diseases. However incontinence rates of up to 100% for neoplasia‐affected dogs have dampened surgeons' enthusiasm for performing prostatectomies. Hypothesizing that incontinence following prostatectomy for prostatic cancer is related to extensive dissection associated with advanced invasion, it is proposed that there exists a subset of dogs with prostatic cancer that require minimal dissection and have the potential for quality survival for pet and owner.
Materials and Methods:  Dogs that received prostatectomy for neoplasia were qualitatively reviewed for commonalities associated with an outcome viewed as successful by owner and veterinarian.
Results:  Early diagnosis (prostatomegaly‐castrated population, high index of suspicion, ultrasound guided cytology) with referral center staging (ultrasound, contrast CT) enabled selection of appropriate surgery (urinary drainage, stent, prostatic enucleation, extrapelvic urethral anastomosis, cystoprostatourethrectomy with ureterocolonic anastomosis) for several dogs.
Conclusions:  A massive demographic shift toward castrated dogs and widespread availability of abdominal u/s can distinguish dogs that may benefit from advanced staging technology and treatment for locally confined prostatic neoplasia.
Application:  Step one is local control. Though based on a dribble of cases, it is clear that some dogs with prostatic cancer can be cured and live high quality lives. We wanted to leak these results so that oncologists will encourage early aggressive investigation and referral for prostatomegaly in castrated dogs.     

The effect of season of the year on the characteristics and composition of dog semen

Semen (collected by digital manipulation) and peripheral blood samples were obtained twice weekly from five fully grown Beagle dogs for a twelve month period from 1st August to 31st July. Sperm concentration, sperm output and glycerylphosphorylcholine concentrations of the second fraction showed evidence of a seasonal variation with highest values recorded in the months of March, April, May and June. The libido of the dogs, volume of ejaculate, percentage dead and abnormal spermatozoa, sperm motility and plasma testosterone concentration showed no evidence of seasonal change.     

Residual effect after salmon oil supplementation on semen quality and serum levels of testosterone in dogs

The objective of present study was to evaluate the effects of oral supplementation of salmon oil on seminal parameters and testosterone serum levels in dogs, following also the residual effects for 60 days after treatment. Nine healthy male dogs with proven fertility, weighing between 10 and 36 kg, ageing from 2 to 11 years, of different breeds, fed diets supplemented with salmon oil at the manufacturer's recommended dosage. The parameters measured were sperm volume, motility, vigour, normal morphology and concentration, live/dead ratio, membrane viability by means of HOST test and serum testosterone levels. Evaluations occurred at baseline (D0), after 90 days of supplementation (D90) and at the end of the experiment, 60 days after supplementation cessation (D150). Results (mean ± SD) obtained at time D0, D90 and D150 were as follows: motility of 76.66% ± 13.7, 92.77 ± 4.41 and 93.0 ± 7.90 (p = .001); normal spermatozoa of 69.11% ± 24.90, 90.00% ± 5.15 and 80.66 ± 16.04 (p = .05); live/dead (%) from 64.44 ± 22.86 to 85.33 ± 8.41 (p = .001); and spermatozoa (%) with integral membrane in the membrane integrity (HOST) test ranging from 76.44 ± 20.74 to 91.22 ± 4.68 (p = .05). Serum levels of testosterone (ng/ml) increased from 5.50 ± 1.13 to 8.84 ± 1.13 at D90 (p = .003) and decreased after 2 months (D150) to 5.13 ± 1.13. In conclusion, a 90‐day supplementation with salmon oil had a positive influence on semen quality and serum testosterone levels. The supplementation of omegas 3 and 6 at the ratio of 10:1 for 90 days determined an increase in concentration and motility of the sperm, and these effects were maintained for 60 days, with the only exception of testosterone levels.     

Effect of supplementation with the reduced form of coenzyme Q10 on semen quality and antioxidant status in dogs with poor semen quality: Three case studies

Oxidative stress owing to an imbalance between reactive oxygen species and antioxidants, such as coenzyme Q10 (CoQ10), is a major contributor to male infertility. We investigated the effects of the reduced form of CoQ10 (ubiquinol) supplementation on semen quality in dogs with poor semen quality. Three dogs received 100 mg of ubiquinol orally once daily for 12 weeks. Semen quality, serum testosterone, and seminal plasma superoxide dismutase (SOD) activity were examined at 2-week intervals from 2 weeks before ubiquinol supplementation to 4 weeks after the treatment. Ubiquinol improved sperm motility, reduced morphologically abnormal sperm, and increased seminal plasma SOD activity; however, it had no effect on testosterone level, semen volume, and sperm number. Ubiquinol supplementation could be used as a non-endocrine therapy for infertile dogs.     

Influence of different anaesthetic protocols over the sperm quality on the fresh,chilled (4°C) and frozen‐thawed epididymal sperm samples in domestic dogs

This study assessed the influence of three different anaesthetic protocols on semen quality obtained from the epididymis. Sixty male dogs undergoing to routine sterilization were assigned to three anaesthetic protocols: thiopental group (TG, n = 20), propofol group (PG, n = 20) and ketamine–dexmedetomidine group (KDG, n = 20). Immediately after orchidectomy, the cauda epididymides and vas deferent ducts were isolated and then a retrograde flushing was performed to collect spermatozoa. In experiment 1, after the initial evaluation of the semen (sperm concentration, sperm motility and the percentages of live spermatozoa, abnormal spermatozoa and acrosome membrane integrity), semen samples were diluted in Tris‐glucose‐egg yolk extender and chilled for 48 hr, and the sperm motility was assessed at 6, 24 and 48 hr. In experiment 2, semen samples were diluted in Tris‐glucose‐egg yolk extender and chilled for 24 hr, and then samples were frozen in two extenders with different glycerol concentrations, to reach a final concentration of 50–100 × 10⁶ spermatozoa ml^?1, 20% egg yolk, 0.5% Equex and 4% and 5% glycerol, respectively. Mean values of total sperm concentration, sperm viability and the percentages of intact acrosome and abnormal spermatozoa were not significantly different between experimental groups, and therefore, the anaesthetic protocols assessed did not affect sperm parameters mentioned above. However, our study confirmed a detrimental effect of the use of thiopental (TG) over the total sperm motility (p < 0.05) and progressive sperm motility (p < 0.05) of the fresh and chilled epididymal sperm samples. The anaesthetic protocols including the application of propofol or ketamine–dexmedetomidine can be used to recover sperm in domestic canids without significant changes in sperm quality compared when semen is collected routinely and these techniques could be applicable to endangered wild canids.     

Seasonal variation in semen quality of bulls and correlations with metabolic and endocrine parameters

Sperm output and semen quality of 17 bulls sampled over 12 months showed minimal output in mid-winter and late summer and minimum quality in late summer. Monthly measurements of luteinising hormone and testosterone concentration in plasma and testosterone concentration in semen were made over 12 months. Serum non-esterified fatty acids (NEFA), albumin and total protein were also measured for the final seven months of this period. Plasma testosterone showed a strong negative correlation with sperm numbers two months hence but not in the current month. Plasma testosterone by bull and semen testosterone by month was also correlated with sperm output. Plasma luteinising hormone three months and serum total protein two months prior was positively correlated with sperm numbers ejaculated and the normality of sperm morphology, possibly by affecting the luteinising hormone dependent A0 to A1 spermatogonial division. NEFA was correlated with initial and post freezing/thawing motility in the current month, possibly by affecting membrane stability. The value of examining the bull in diagnosing infertility of cows where nutritional stress may have occurred is suggested, as is the use of albumin/total protein and NEFA measurements as a prognostic aid for time to return to normality of function of such bulls.     

Seminal Plasma Characteristics and Expression of ATP‐binding Cassette Transporter A1 (ABCA1) in Canine Spermatozoa from Ejaculates with Good and Bad Freezability

The composition of seminal plasma and the localization of the ATP‐binding cassette transporter A1 (ABCA1) in spermatozoa from good and bad freezers were compared to frozen–thawed spermatozoa from the same dog. Ejaculates were obtained from 31 stud dogs, and the sperm‐rich fraction (SRF) was kept for analysis. One aliquot was used for the analysis of concentration, progressive motility (P; CASA), viability (V; CASA) and leucocyte count, and the analysis was performed by flow cytometry (FITC‐PNA/PI), SCSA and HOST. In seminal plasma, concentration of albumin, cholesterol, calcium, inorganic phosphate, sodium, potassium, zinc and copper was measured. Semen smears were prepared and evaluated for the expression of ABCA1. The remainder of each ejaculate was frozen. After thawing, the quality assessment was repeated and further smears were prepared. According to post‐thaw semen quality, dogs were assigned to good freezers (n = 20) or bad freezers (n = 11), the latter were defined as < 50% progressive motility and/or > 40% morphologically abnormal sperm and/or < 50% viability. Bad freezers were older than good freezers (5.3 vs 3.4 years, p < 0.05). In bad freezers, the percentage of sperm with ABCA1 signal in the acrosome was lower (26.3% vs 35.7%, p < 0.01) and the percentage of sperm with complete loss of ABCA1 signal higher (46.7% vs 30%, p < 0.01); the percentage of dead spermatozoa was higher (36.1% vs 25.5%, p < 0.05), and the concentration of cholesterol and sodium in seminal plasma was lower than in good freezers (p < 0.05). We conclude that in thawed bad freezer sperm, an increase in acrosome damages coincided with an increased loss of cholesterol transporters and cell death, and a lower cholesterol concentration in seminal plasma. Follow‐up studies revealed whether a relation exists between these findings.     

Serum testosterone and estradiol 17-beta concentrations in 15 dogs with perineal hernia

Serum testosterone and estradiol 17-beta concentrations, and serum testosterone-to-estradiol ratio were evaluated in 15 dogs (greater than or equal to 5 years old) with perineal hernia (9 sexually intact males and 6 castrated males) and in 9 clinically normal sexually intact male dogs greater than or equal to 5 years old. There was no significant difference in serum testosterone-to-estradiol ratio between sexually intact male dogs with perineal hernia and clinically normal sexually intact male dogs. In castrated dogs with perineal hernia, serum testosterone concentration and testosterone-to-estradiol ratio were significantly (P less than 0.05) lower, compared with those values in sexually intact dogs with perineal hernia and in clinically normal sexually intact male dogs. There was no significant difference in serum estradiol 17-beta concentration among sexually intact male dogs with perineal hernia, castrated dogs with perineal hernia, and clinically normal sexually intact male dogs. Serum testosterone and estradiol 17-beta concentrations in dogs with perineal hernia did not differ from those values in clinically normal male dogs of the same age. Castration cannot be recommended for the treatment of perineal hernia unless a castration-responsive contributing factor such as prostatomegaly is identified, unless the pelvic diaphragm of dogs with perineal hernia has high sensitivity to normal or low serum testosterone and estradiol 17-beta concentrations, or unless there is documentation that other androgens and/or estrogens are involved.     

Effects of Seminal Fluid Fractions on Plasma and Acrosome Membrane Integrity and Mitochondrial Membrane Potential Determined by Flow Cytometry in Chilled Canine Spermatozoa

Depending on the mammal species, the use of seminal plasma during semen processing for cryopreservation has been found to have both beneficial and detrimental effects. This study was designed to determine the effects of the second (SF) and third [prostatic fluid, (PF)] ejaculate fractions on plasma membrane and acrosome integrity, mitochondrial membrane potential (ΔΨm), phosphatidylserine (PS) translocation and sperm motility in chilled canine spermatozoa by flow cytometry. After pooling the second sperm‐rich fraction of ejaculates from six dogs, samples for each assay were preserved at 5°C for 72 h in egg yolk‐TRIS extender (EYT) alone (control) or supplemented with seminal fluid from the second (EYT‐SF) or third (EYT‐PF) ejaculated fractions. After cold storage, groups EYT‐SF and EYT‐PF showed significantly higher percentages of sperm cells with an intact acrosome [68.8 ± 1.4%, 69.6 ± 2.6% (p < 0.01)] and intact plasma membrane [48.1 ± 2.8%, 50.4 ± 8.2% (p < 0.001)] than that observed in EYT [51.7 ± 3.2% and 33.3 ± 4.1% respectively]. Only in EYT‐SF was PS translocation significantly reduced compared to EYT‐PF and EYT [3.9 ± 0.4%, 10.2 ± 2.2% and 9.0 ± 1.5%, respectively (p < 0.001)]. However, significantly diminished sperm motility was observed in EYT‐SF and EYT‐PF compared to EYT [36.8 ± 2.1%, 35.5 ± 2.3% and 78.4 ± 4.7% (p < 0.001)]. No significant differences were detected in ΔΨm (p > 0.05). In conclusion, supplementing semen extenders with seminal fluid from the second or third fractions of the ejaculate supplementation helps to preserve the integrity of the plasma and acrosome membranes along with the mitochondrial membrane potential but seems to compromise the motility of canine spermatozoa chilled for 72 h.     

Endogenous serum insulin concentration in dogs with diabetic ketoacidosis

Objective: To determine endogenous serum insulin concentration in dogs with diabetic ketoacidosis (DKA), and to compare it to endogenous serum insulin concentration in diabetic dogs with ketonuria but no acidosis (KDM), diabetic dogs with uncomplicated diabetes mellitus (DM) that did not have ketonuria or acidosis, and dogs with non‐pancreatic disease (NP). Design: Prospective study. Setting: Veterinary Hospital of the University of Pennsylvania. Animals: Forty‐four client‐owned dogs; 20 dogs with newly diagnosed diabetes mellitus (7 dogs with DKA, 6 dogs with KDM, and 7 dogs with DM) and 24 dogs with non‐pancreatic disease. Interventions: Blood and urine samples were obtained at the time of admission to the hospital. Measurements and main results: Signalment, clinical signs, physical examination findings, and concurrent disease were recorded for all dogs. Blood glucose concentration, venous blood pH, venous blood HCO3^? concentration, urinalysis, and endogenous serum insulin concentration were determined in all dogs. Dogs with DKA have significantly decreased endogenous serum insulin concentrations compared to dogs with DM (P = 0.03) and dogs with non‐pancreatic disease (P = 0.0002), but not compared to dogs with KDM (P = 0.2). Five of 7 dogs with DKA had detectable endogenous serum insulin concentrations, and 2 of these dogs had endogenous serum insulin concentration within the normal range. Conclusions: Diabetic dogs with ketoacidosis have significantly decreased endogenous serum insulin concentration compared to dogs with uncomplicated diabetes mellitus. However, most dogs with DKA have detectable endogenous serum insulin concentrations, and some dogs with DKA have endogenous serum insulin concentrations within the normal range.     

Effects of Incubation Temperature and Semen Pooling on the Viability of Fresh,Chilled and Freeze‐Thawed Canine Semen Samples

This study assessed the effects of different incubation temperatures on semen viability and the influence of pooling on semen longevity. In experiment 1, semen samples were collected from five dogs, individually processed (individual semen: IS) and then aliquots from each male were pooled (pooled semen: PS). Semen samples (IS and PS) were diluted in a Tris‐glucose‐yolk extender and preserved as fresh (37 and 25°C) and chilled semen (4°C). Sperm motility and the percentages of sperm abnormalities and acrosome membrane integrity were assessed for 24 h. Storage at 25 or 4°C for the first 24 h yielded similar semen quality, but incubation at 37°C caused drastic reduction in sperm motility from 8 h of incubation onwards. In experiment 2, the semen was processed in the same way to that of experiment 1 and then preserved at 25 or 4°C until semen inactivation. Semen that was incubated at 25°C became completely inactive after 3–4 days of storage, while semen that was preserved at 4°C presented with more gradually decreased sperm motility (mean values of 40–60% for the first 8 days). In addition, the mixing of semen was only observed to influence the sperm quality of the samples stored at 4°C. In experiment 3, semen was collected from five dogs, pooled and frozen in liquid nitrogen; after thawing, it was preserved at 37, 25, 15 and 4°C, and the sperm quality was defined. The motility of the freeze‐thawed semen samples decreased quickly in the first 4 h after thawing, regardless of the preservation temperature of the thawed semen. This study confirmed that semen preserved at 37°C should be used within a maximum of 12 h, while the semen stored at 25°C shows acceptable quality for 24 h. Chilled semen presented highest most sustainable quality, especially when semen is processed as pooled semen.     

Effect of Ascorbic Acid Concentrations,Methods of Cooling and Freezing on Boer Goat Semen Cryopreservation

To improve the Boer goat semen quality during cryopreservation process, three experiments were carried out to investigate the effect of (i) different concentration of ascorbic acid supplementation (ii) rate of cooling with chilled semen characteristics and (iii) method of freezing on post‐thaw Boer goat sperm using Tris‐based extender. Ascorbic acid at 8.5 mg/ml improved the sperm parameters (motility, integrity of membrane and acrosome, morphology and viability), compared to control in cooled samples (p < 0.05). With regard to other concentrations and post‐thawed parameters, ascorbic acid at 2.5–8.5 mg/ml led to higher percentages of sperm motility and integrities of membrane and acrosome when compared to control (p < 0.05). Slow cooling rises to higher percentages of sperm motility, acrosome integrity and viability, in comparison with fast cooling, in terms of cooled and frozen samples (p < 0.05). Programmable freezing method produced the higher percentages of sperm motility, integrities of membrane and acrosome and viability when compared to the freezing method of polystyrene box during goat sperm freezing (p < 0.05). In conclusion, chilled and post‐thawed sperm quality of Boer goat was improved when a Tris‐based extender supplemented with ascorbic acid was used at stages of different cooling rates and freezing methods.     

In vitro evaluation of cryopreserved bovine sperm and its relation to field fertility in fixed‐time artificial insemination

This study aimed to assess characteristics of bovine cryopreserved sperm and evaluate its relation to field fertility in fixed‐time artificial insemination (FTAI). Semen samples of 16 bulls were used to inseminate 811 Nellore cows, and four of these bulls were also used to inseminate 101 Nellore heifers. Samples of the same ejaculate used for FTAI from each bull were analysed in the laboratory after thawing. Sperm motility and vigour were subjectively assessed by light microscope, and integrity of the plasma and acrosome membranes, and H₂O₂ production were evaluated by flow cytometer. Relation among sperm characteristics and pregnancy rate of cows and heifers were evaluated by univariate and multivariate logistic regression. Subjective sperm motility and vigour did not affect the probability of pregnancy in cows or heifers. In univariate analysis for pregnancy in cows, sperm traits related to acrosome injury positively affected probability of pregnancy mainly when associated with plasma membrane integrity; H₂O₂ production seems to be less important than plasma membrane integrity in affecting probability of pregnancy. In multivariate analysis, sperm traits related to injured acrosome positively affected probability of cow and heifer pregnancies while intact acrosome was negatively related to cow pregnancy. Intact plasma membrane and high H₂O₂ production were positively related to cow pregnancy but negatively related to heifer pregnancy. Results suggest that a capacitation‐like status of the acrosome may benefit probability of pregnancy in cows.     

Assessment of Different Sperm Quality Parameters to Predict in vitro Fertility of Bulls

Frozen‐thawed semen from six bulls with high (> 60%) and low (20–35%) in vitro fertility was used for studying the predictive value of simple sperm quality tests with respect to in vitro fertilization (IVF) outcome as assessed by pronucleus (PN) formation ability. Sperm quality parameters, such as sperm concentration, motility, progressive motility, live‐dead sperm ratio, morphology, membrane integrity, mitochondrial activity and acrosomal status were analysed using both conventional and automatic techniques at three time points during the IVF process, namely after sperm thawing, Percoll differential gradient centrifugation and IVF. Associations between the sperm quality parameters before and after IVF, and PN formation ability were assessed by using linear regression analyses. The percentages of motility, progressive motility and normal morphology determined after sperm thawing, and the percentage of live spermatozoa assessed after Percoll preparation by using nigrosin‐eosin (N‐E) staining showed a good correlation with PN formation ability, but the regression parameters were borderline not significant. These parameters formed the most reliable basis for predicting IVF outcome. After IVF, the percentage of live spermatozoa determined by using N‐E staining was the only sperm quality parameter showing a significant association with the PN formation ability of a given bull. This sperm quality test can be used as a non‐invasive method to estimate the PN formation ability of oocytes which are further cultured to assess embryonic development.     

Canine prostate specific esterase (CPSE) as an useful biomarker in preventive screening programme of canine prostate: CPSE threshold value assessment and its correlation with ultrasonographic prostatic abnormalities in asymptomatic dogs

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.