Prostate vascular flow: The effect of the ejaculation on the power doppler ultrasonographic examination.

Power Doppler sonography (PD) can accurately depict tissue perfusion, recognize slow flows, and is relatively angle independent. The monitoring of local blood flow by Doppler ultrasonography is helpful in differentiating prostatic physio-pathological conditions, but the recognizing of physiological variables that could affect it is crucial to apply this technique in clinical practice. This study aimed to evaluate if ejaculation affects blood flow to the prostate and to state how long this effect lasts. Serial PD examinations of prostate were performed in 18 dogs (1-5 years, 6-40 kg) immediately before (T0) and after (T1) the ejaculation, and repeated 6 (T2), 18 (T3) and 24 (T4) hours later. For each examination, two representative PD images were chosen and ranked by two independent observers according to the following scoring system: 0 = mild subcapsular (S) vascularization without clear evidence of parenchymal (P) vascularization; 1 = moderate P and S vascularization; 2 = severe S and moderate P vascularization; 3 = severe P and moderate S vascularization; 4 = severe P and S vascularization. Interobserver agreement was assessed using Kappa of Cohen. Ranked data, grouped according to time, were compared by ANOVA and Tukey HSD test (p < .05). Variations in the vascular flow pattern at different times were observed for all dogs. The statistical analysis evidenced a significant difference for T0 vs T1 and vs T2 and vs T3 (p < .01), with no significant difference for T0 vs T4 (p > .05). Interobserver agreement was very good (Kappa of Cohen = 0.86). This study demonstrated a definite increase in vascular flow to the prostate after ejaculation. The present results suggest a minimum of 24 hr sexual rest before the PD examination of the gland. This result should be taken into account whenever Doppler sonography is used to evaluate potential hyperaemia in dogs suspected of having prostate abnormalities.

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.

Differential surface glycoprofile of buffalo bull spermatozoa during mating and non-mating periods.

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 NeuNAcα2-3Galß1,3(±NeuNAcα2-6)GalNAc, Galß1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galß1,3/4GlcNAcß1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galß1,4GlcNAc, 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.

Therapeutic Ultrasound as a Potential Male Dog Contraceptive: Determination of the Most Effective Application Protocol.

Ultrasound is one of the most promising forms of non-invasive contraception and has been studied in several animal models. The objective of the current investigation was to determine the most practical and effective application protocol for dog sterilization. A total of 100 dogs were divided into five equal groups. Group A received 5-min applications three times performed at 48-hr intervals and covering the entire testicular area at frequency of 1 MHz; Group B received 5-min applications three times performed at 48-hr intervals over the dorso-cranial area of the testis at frequency of 3 MHz; Group C received three sequential 5-min applications (at 5-min intervals between applications) covering the entire testicular area at frequency of 1 MHz; Group D received 15-min applications two times performed at 48-hr intervals and covering the entire testicular area at frequency of 1 MHz. The experimental groups' ultrasound had an intensity of 1.5W/cm(2) . The Control Group had the same procedure as Group A, but with the transducer switched-off. Dogs were surgically castrated 40 days following the treatment for histological examination. Azoospermia, testicular volume reduction and apparently irreversible testicular damage were achieved by Group A. No effects were noticed in the other groups. Testosterone levels remained within physiological range with all application protocols. A regimen of three applications of ultrasound at 1 MHz, and 1.5 W/cm(2) , lasting 5 min with an interval of 48 h was effective as permanent sterilization in the dog without hormonal impact.

A rare case of simple syndactyly in a puppy.

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.

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.

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.

Influence of feeding level on live body weight and semen characteristics of Sardinian rams reared under intensive conditions.

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 control-concentrate (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.

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.

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.

Induction of fertile oestrus in cats by administration of hCG and calcium-naloxone.

The aim of this study was to evaluate the possibility of inducing fertile oestrus in queens by administering hCG in combination with Ca(2+)-naloxone. It is well established that an increase in endogenous opioids leads to a decrease in LH. The administration of naloxone, an opioid antagonist, inhibits endogenous opioidergic tone and induces the onset of pro-oestrus. The opioidergic block is related to the increase in binding of beta-endorphins to specific receptors, which determines calcium channel blockage. Pretreatment with hCG results in a rapid increase in the number of LH receptors and Ca(2+)-naloxone induces G protein activity. Twenty-one anoestrous queens were divided into four groups: (i) group 1, nine queens were treated with a single s.c. injection of hCG (1000 iu) and daily for 4 days with 0.1 ml kg-1 body weight i.m. of a solution containing 0.4 mg naloxone ml-1 dissolved in 20% calcium gluconate; (ii) group 2, four animals were treated with a single s.c. injection of hCG (1000 iu); (iii) group 3, four queens were treated with Ca(2+)-naloxone (0.1 ml body weight kg-1 i.m.) daily for 4 days; and (iv) group 4, four queens received no treatment (controls). Queens were monitored using vaginal cytology and blood progesterone concentrations, and pregnancy was detected using ultrasonography. In groups 2, 3 and 4 clinical signs of oestrus were not observed. In group 1, 88.8% of treated queens were mated (8 of 9) and ovulated on the basis of an increase in progesterone, and 75% (6 of 8) of these queens became pregnant. In conclusion, pretreatment with hCG increased the number of LH receptors and Ca(2+)-naloxone antagonized the hypothalamic GnRH opioid block thus inducing the pulsatility of LH leading to fertile oestrus in queens.