Application of B-mode ultrasonography in the assessment of the dog penis

With the purpose to characterize the ultrasonographic features of the dog penis, 20 healthy dogs aged from 2 to 11 years old were used. Ultrasonography was performed using a 5-9MHz linear probe and a Philips HD3 scanner. Ultrasonograms were obtained at the penile body (pre- and post-scrotal sections) and at both the bulbus glandis and the pars longa glandis, with the dog placed in dorsal recumbency. The typical anatomy of the dog penis is easily demonstrated in ultrasonographic images, as revealed in this study. On ultrasonograms taken at the body level, the two corpora cavernosa showed a homogeneous, hypoechoic structure; ventrally, the corpus spongiosum appears as a slightly hyperechoic structure surrounding the urethra. In glans ultrasonograms, the most typical feature is the os penis, observed as a strong hyperechoic structure, whose acoustic shadow impairs the screening of adjacent tissues. Lining on its concavity, the corpus spongiosum is observed as a slightly hyperechoic tissue around the urethra. An ovoid, regularly hypoechoic tissue corresponding to the erectile tissue of the pars longa, surrounds these structures. The bulbus glandis is visualized as a regular hypoechoic structure with small anechoic areas, corresponding to sinusoidal spaces. The albuginea is evidenced as a well-defined, hyperechoic external layer. The preputial cavity appears as a thin anechoic layer surrounding the albuginea. The results of this study demonstrate that ultrasonographic assessment of the dog penis is easily performed and accurate enough for penile examination. Furthermore, ultrasonography of the dog penis can become an interesting diagnostic tool to explore in the canine practice, during breeding soundness evaluation and also whenever penile exposition is impossible to achieve.     

Trehalose enhanced the freezability of Poodle dog sperm collected by an artificial vagina (AV)

In an attempt to develop a suitable freezing method for Poodle dog sperm, an experiment was conducted to investigate semen collection methods of digital stimulation and an artificial vagina (AV), using Tris and trehalose-egg yolk extender, on the characteristics and cryopreservation of sperm. Two dogs (dogs A and B) were subjected to semen collection by digital stimulation and AV. The volume, sperm concentration, sperm motility index (SMI) and acrosome status of ejaculates were determined immediately after collection. The remainder was frozen as pellets in Tris and trehalose-egg yolk extender. Sperm motility index was evaluated after thawing and during a thermal resistance test, and acrosome integrity was also assessed. No significant differences regarding sperm concentrations, SMI and acrosome integrity were observed between semen collected by AV and digital stimulation. However, when dog sperm were collected by an AV and frozen in trehalose-egg yolk extender, the motility index of frozen-thawed sperm was significantly improved compared to sperm frozen in Tris-egg yolk extender which were collected by digital stimulation. In conclusion, semen collected by an AV and frozen in trehalose-egg yolk extender was effective in enhancing the freezability of Poodle dog sperm.     

Effect of semen collection method on sperm motility of gray wolves (Canis lupus) and domestic dogs (C. l. familiaris)

Genetic management of Mexican gray wolves includes semen banking, but due to the small number of animals in the population and handling restrictions, improvements in semen collection and cryopreservation rely on results from studies of domestic dogs. Semen collection from wolves requires anesthesia and electroejaculation, which introduce potentially important variables into species comparisons, as dog semen is typically collected manually from conscious animals. To investigate possible effects of collection method on semen quality, we compared semen collection by the traditional manual method and by electroejaculation (EE) in a group of dogs (n = 5) to collection by EE only in wolves (n = 7). Samples were divided into two aliquots: neat or diluted in Tris/egg yolk extender, with motility evaluated at intervals up to 24 h. There were no differences (P > 0.10) in sperm motility in either neat or extended samples at 24 h from EE dogs and wolves, although motility of the wolf neat samples declined more rapidly (P < 0.05). However, there were differences (P < 0.01) between EE and manually collected dog semen in motility at 24 h, in both the neat and extended samples. Therefore, general motility patterns of dog and wolf semen collected by EE were similar, especially when diluted with a Tris/egg yolk extender, but sperm collected from dogs by EE did not maintain motility as long as manually collected samples, perhaps related to the longer exposure of EE samples to more prostate fluid.     

L’erection et l’ejaculation psychogenes: Mythe ou realite? Intérêt de leur recherche chez le blessé médullaire

Erection and ejaculation can be induced by two forms of stimulation. The first is mechanical: touching of the penis, scrotum and anus stimulates the pudendal nerves and a reflex erection appears via the parasympathetic pathway. The second form of sexual stimulation is psychological: erotic pictures and fantasies stimulate the splanchnic nerves and the hypogastric plexus, via the orthosympathetic dorso-lumbar pathway, and induce a psychogenic erection. Spinal injury can destroy a part of these neurological routes and cause impotence and/or failures of ejaculation. We have studied 35 patients reporting a sexual dysfunction after a spinal injury, and have tried to generate an erection and ejaculation first with mechanical stimulation, and then with erotic pictures without mechanical genital stimulation. Tumescence was registered using a plethysmograph and semen was collected in order to study spermatogenesis. The spinal cord was destroyed in 12 patients, but orthosympathetic stimulation produced a psychogenic erection in 15 cases, followed by ejaculation in six cases. This form of stimulation seems to be better than prostaglandin or electrical stimulation.     

Avulsion of the bovine prepuce from its attachment to the penile integument during semen collection with an artificial vagina

Avulsion of the prepuce (AP) from the penile integument during intromission into an artificial vagina (AV) is the most common injury involving the penis and prepuce of bulls collected for the production of frozen semen at artificial insemination (AI) centers. We observed clinical cases at two AI centers. Because temporary loss of seminal collections from genetically superior bulls occurs and the possibility of impotency exists, the economical loss from AP is significant. Surgery, rather than a nonsurgical procedure, is the preferred method for repair of AP if the diagnosis occurs on the same day as the injury. After injury, seminal collection is avoided. The bull can then begin a conditioning program prior to returning to a normal semen collection schedule. A specific etiology for AP remains undetermined; however, many variables potentially involved in its occurrence are known. Bulls with good libido following 4 to 5 yr of sexual inactivity are particularly susceptible to AP. No genetic predisposition was observed. At present, we believe that AP is caused by many factors and that implementing preventive measures does help.     

Stallion ejaculation induced by manual stimulation of the penis

This paper reports the use of a procedure for collection of semen from stallions by manual stimulation of the penis while the stallion is standing. Our use of this method with 18 stallions of various ages and types of semen collection experience indicates that this method may be an efficient alternative to traditional semen collection techniques using an artificial vagina and stimulus mare or dummy mount mare. Our observations, together with those of others who have tried the manual technique, suggest that both animals and handlers can be readily trained to use this method. Limited data suggests that semen samples obtained by manual stimulation are similar to those obtained using an artificial vagina.     

Etude de l’erection chez le rat: un nouveau modele physiologique

Penile erection is a muscular and vascular event mediated by the autonomic nervous system. The neurophysiology of erection remains poorly understood and controversial, requiring a suitable model for in-vitro studies of erectile function. Such a model, based in the rat whose penile innervation is very similar to man, is described here. The first study using this model considers the influence of systemic blodd pressure (BP) on penile erection. In 33 anaesthetized rats the pelvic and cavernosal nerves were identified and dissected. Supra maximal electrical stimulation was delivered over 1 minute by a train of 1 ms pulses onto the pelvic nerve (10 V, 15 Hz) or the cavernosal nerve (6 V, 10 Hz). Systemic blood pressure and intracavernosal pressure (ICP) were monitored and stored on a computer. As in previous animal models (dog, monkey), four phases of the cavernosal response to neural electrical stimulation were observed: latency, tumescence, full erection, and détumescence. In all rats electrical stimulation of either the pelvic or cavernosal nerves significantly increased intracavernosal pressure. Complete erectile response (rigidity and unfolding of the penis) was only seen with intracavernosal pressures > 95 mm Hg. Intracavernosal pressure increased proportionally with blood preessure during the full erection phase according to the equation ICP=0.94 BP ? 31 mm Hg (r=0.94 BP ? 31 mm Hg (r=0.94) for electrical stimulation of the cavernosal nerve, or the alternative aquation ICP=0.76 BP ? 21 mm Hg (r=0.73) for electrical stimulation of the pelvic nerve. The rat is a readily available model for the study of erection and present obvious advantages over existing models such as the dog, cat and monkey. Cavernosal repsonse to neural stimulation was closely related to arterial blood pressure and the two linear equations presented above should be considered further in studies modifying autonomic neurotransmission as well as in relation to the effects of pharmacological compounds with vasomotor actions on erectile function.     

Preliminary analysis of reproductive,behavioral and physiological characteristics of military working dogs

The use of dogs in military work environments has always aroused great interest in the general population and determining the stress levels they go through is extremely important to maintain their welfare. The aim of this research was to evaluate if the work shifts in military working dogs leads to stress conditions and if this working influences on the reproductive performance and life quality. The study was conducted at the Military Police Kennel located at Campinas, Sao Paulo, Brazil. Eight male dogs of four different breeds (German Shepherd, Belgian Malinois Shepherd, Doberman, and Rottweiler) were evaluated during two different shifts: Working Shifts: animals working 12 hours a day with 2 hour-interval; and Control Shifts: animals that were on their day off (36 hours). Saliva samples were collected for cortisol analysis at the control and working shifts. The study was carried out over 60 days and analyzed behavior, physiology, and reproduction quality. Saliva samples, behavior observation of stereotyping, resting and moving activities and semen analysis were collected by digital stimulation (for combined second and third fractions). The salivary cortisol levels during the control and working shifts were between 0.361–0.438 and 0.312–0.592 µg/dL, respectively; the highest values were found at the end of working shifts. The animals were resting during most of the observation period, but few showed stereotypic behaviors. The testicular consistency was firm and semen parameters were within the normal values in German Shepherd, Belgian Malinois Shepherd, and Doberman dogs. However, Rottweiler dogs had a higher rate of sperm abnormalities, higher salivary cortisol levels, and more stereotypic behaviors. Nevertheless this work highlights the importance of further research relating reproduction and cortisol levels in military dogs.     

Heritability of semen characteristics in dogs

Retrospective analysis was performed on semen collected from 24 dogs (parents: 14 Labrador retrievers and 10 Golden retrievers) aged between 16 and 28 months of age. The dogs were part of a large breeding programme but lived in the homes of volunteer families. The semen was subjected to a standardised examination procedure including assessment of: percentage normal motility, sperm concentration, total sperm output, percentage of live normal sperm, and total number of live normal sperm. Semen was subsequently collected from one son of each of the parents when the offspring were aged between 16 and 28 mo (offspring: 14 Labrador retrievers and 10 Golden retrievers), and was subjected to the same examination procedures conducted by the same technician. Examination of breeding records demonstrated that each of the 48 dogs achieved at least one pregnancy within a period of 3 months before to 3 months after the semen collection.There was a weak correlation between parents and offspring for each of the 5 semen parameters, although none of these were statistically significant. Narrow sense heritability measures were low for all parameters except for the heritability of high sperm motility (rN2 = 0.57) and the heritability of low total sperm output (rN2 = 0.57).It is plausible that breeding selection procedures may be useful in dog breeding programmes in an attempt to improve semen quality, although any impact upon fertility is yet to be proven.     

A method for collecting semen by fingers in the African green monkey (Cercopithecus aethiops) and properties of the semen collected

We tried to make a successful collection of semen by human fingers, using ten African green monkeys (Cercopithecus aethiops) aged from 5-7 years, which were bred and reared at Tsukuba Primate Center for Medical Science. Under anesthesia, the animals were laid side way and their penis were gently stimulated by fingers. Semen was successfully obtained from five animals aged 6-7 years, but it was impossible to collect semen from the remaining five animals aged 5 years. It took only 5-15 seconds from the beginning of stimulation to the end of semen collection. The volume of semen collected, the number of spermatozoa and spermatozoal motility rate averaged 1.5 ml, 3.1 x 10(8)/ml, and 46%, respectively.     

Semen quality in dogs and the influence of a short-interval second ejaculation

Semen quality was examined in each of 65 known fertile dogs. Values were found to be similar to those previously published, although an apparent breed influence was demonstrated, with German shepherd dogs producing ejaculates of larger volume and greater total spermatozoal output than other breeds. A second ejaculate was collected from each dog with a mean interval of 63 min. The second samples had significantly lower values for the volume of the second fraction, the spermatozoal concentration and the total spermatozoal output. There were no differences for the percentage motility or the percentage of morphologically normal live spermatozoa. While there was no increase in semen quality of the second ejaculate, the technique may be useful since it results in the collection of approximately 70% more spermatozoa than a single ejaculate. These spermatozoa also had normal motility and morphology and could, therefore, be used for insemination or cryopreservation.     

Effect of the GnRH antagonist, acyline, on canine testicular characteristics

The primary objective was to assess the effects and clinical safety of a single high-dose of the third generation GnRH antagonist, acyline, on testicular characteristics in male dogs. Seven dogs were followed up weekly for six, 2-week periods (-2, -1, 1, 2, 3 and 4). At the end of the second period, they were given acyline (330 microg/kg sc). Responses to treatment varied among individual animals. Testicular consistency and scrotal diameter were slightly diminished (P>0.05) in Periods 1, 2, and 3. Libido and erection decreased during Periods 1 and 2 (P<0.05). Second and third fractions of the ejaculate volume, sperm count and motility varied throughout periods (P<0.01); there was a clear impairment of these parameters (< or =0.2 mL, < or =0.6 mL, < or =0.5 million/mL and < or =30%, respectively) around the second week of Period 1, followed by slow improvement (to the end of the study). Semen volumes and motility diminished during Period 1 (P<0.05). Sperm count decreased during Periods 1, 2, and 3, relative to Periods -2 and -1 (22.7+/-11.7, 62.8+/-19.9, and 51.0+/-25.4 versus 235.7+/-63.3 and 315.5+/-27.3, respectively; P<0.05; (L.S.M.+/-S.E.M.). Morphologically abnormal sperm increased during Periods 2 and 3 (up to 64.9+/-2.6%; P<0.05). Throughout the study, no dog had hematological, biochemical, local, or systemic side effects. In conclusion, a single high-dose acyline treatment severely decreased semen quality with no adverse effects.     

Penile paralysis and paraphimosis associated with reserpine administration in a stallion

Reserpine was administered to an 8-yr-old Thoroughbred stallion at a dosage of 5 mg subcutaneously (s.c.) every 2 wk for a 2-mo period to control unmanageable behavior. Reserpine produced a satisfactory calming effect that lasted for about 2 wk. After the last injection, the stallion developed penile paralysis and was unable to retract his penis, resulting in paraphimosis and attendant penile edema. The prolapsed penis was reduced and kept within the prepuce by placing a purse string suture in the preputial orifice. Phenylbutazone was given orally (1 gm) and the stallion was exercised for 20 to 30 min twice daily. This treatment was continued for 20 d with little improvement. The purse string retention suture cut through the skin 5 d after the stallion was discharged from the clinic. The penis was then supported against the abdominal wall and the stallion was exercised by hand for 30 min each day. The stallion was not used for breeding within 34 mo after the last injection of reserpine. A breeding soundness examination was performed approximately 3 yr after the initial injury. At this time the stallion's penis was noted to extend 5 to 8 cm from the prepuce when in a detumescent state. Although the stallion protruded his penis when exposed to a mare in estrus, a full rigid erection was never attained. Examination of the penis revealed partial engorgement of the corpus cavernosum penis and a 2.5-cm-wide dorsal semi-circumferential depression of the penile shaft approximately 10 to 12 cm proximal to the glans penis. The penile shaft and glans penis distal to this depression were cooler than the proximal portion of the penis. Semen collection was attempted, aided by manual insertion of the penis into the artificial vagina. When serving the artificial vagina, no "belling" of the glans penis was observed, although ejaculation occurred. Semen evaluation indicated normal spermatozoal motility and morphology parameters. The stallion was able to breed several mares with manual assistance to guide the penis into the vagina and one mare was diagnosed pregnant.     

Surgical treatment of balanitis xerotica obliterans

Balanitis xerotica obliterans, or kraurosis penis, is a chronic progressive scleroatrophic process of the penis, prepuce, and urethral meatus. This syndrome is due to lichen sclerosus et atrophicus of the genital region. We have observed 32 patients, whose ages ranged from 24 to 78, with different clinical and pathologic findings. Clinical symptomatology consisted of painful erection with secondary impotence, burning, itching, and urinary disorders. The treatment in the early stages is pharmacologic; stenosis of the meatus, phimosis, scar adhesions, fissures, and erosions of glans and prepuce prescribe a surgical treatment. We have performed modified circumcision, meatotomy and meatoplasty, removal of the scleroatrophic tract and subsequent grafting. The functional results were satisfactory.     

Semen quality during vincristine treatment in dogs with transmissible venereal tumor

The aim of this study was to evaluate the direct effects of vincristine on semen quality in dogs with transmissible venereal tumor (TVT). We examined the semen of 17 dogs suffering from TVT during vincristine treatment. Each animal received 0.6 mg, i.v. vincristine sulphate per square meter of body surface, per week for 4 wk until complete regression of the tumor. The following semen parameters were evaluated: semen volume (second fraction), sperm concentration, total spermatozoa per ejaculate, percentage of progressively motile spermatozoa, percentage of dead spermatozoa, percentage of swollen spermatozoa (hypo-osmotic swelling test) and percentage of morphologically abnormal spermatozoa (primary and secondary defects). Semen was collected and evaluated prior to the beginning of treatment, 3 d after each vincristine injection and 15 d after the last injection. Semen characteristics transiently deteriorated during treatment, but returned to normal 15 d later. These changes were attributed to a direct effect of vincristine on the extragonadal spermatozoal reserves contained in the epididymis and ductus deferens. A GnRH stimulation test was also performed after each semen collection in order to assess the function of the hypothalamic-pituitary-Leydig cell axis. No effect was noted on the above axis.     

Retrograde ejaculation occurs in the dog, but can be prevented by pre-treatment with phenylpropanolamine: a urodynamic study

Retrograde ejaculation is partial or total propulsion of semen from the posterior urethra into the urinary bladder; it is well characterized (and relatively common) in humans, with only a few reports in animals. Our objectives were to determine whether retrograde flow of semen occurred during ejaculation in mature dogs with normal fertility, and to determine the effects of phenylpropanolamine on this phenomenon (dose-titration, switch-back study). Retrograde ejaculation and urethral pressure profile measurements were evaluated (double-blind) in six dogs after 5 days of oral treatment with phenylpropanolamine (0, 2, 4, or 8mg/kg); all dogs received all treatments (at 2-week intervals). The number of sperm in the urine was determined before and after each manual sperm collection. Urethral pressure profiles were obtained three times during each procedure. In the absence of phenylpropanolamine, sperm were present in the bladder after semen collection in all dogs (number varied significantly among individuals). The mean (+/-S.D.) number of sperm in the bladder was 17.0+/-5.0, 18.5+/-1.2, 5.1+/-5.0, and 4.8+/-0.1x10(6) sperm for 0, 2, 4, and 8mg/kg, respectively (no significant difference between dogs given 4 or 8mg/kg, but both were significantly lower than those given 0 or 2mg/kg). This reduction was significantly correlated to the increase in mean urethral pressure at the level of the sphincter (39cm versus 59cm H(2)O in placebo-treated dogs versus those given 8mg/kg). In conclusion, we confirmed that retrograde ejaculation occurred during the ejaculatory process in normal dogs, and we demonstrated that phenylpropanolamine (4 or 8mg/kg once daily for 5 days before collection) increased urethral pressure and reduced the number of sperm voided into the bladder during ejaculation.     

Breeding soundness examinations of rams and bucks, a review

Breeding soundness examinations of rams and bucks may be performed by veterinarians as a service for clients. A physical examination for breeding soudness includes a general examination for health with special consideration of the reproductive organs. Careful examination of the penis, prepuce, and testicles may reveal diseases or abnormalities which decrease reproductive potential. Rams with small or hypoplastic testicles are unsatisfactory potential breeders as testicle size is positively related to sperm production. Libido may be assessed during semen collection procedures or from observations of the owner. Semen quality may be evaluated using tests for motility, concentration, and morphology of spermatozoa. Rams may be declared satisfactory, questionable, or unsatisfactory potential breeders as a result of the examination.     

Characterization of semen collected from beagles and captive Japanese black bears (Ursus thibetanus japonicus)

This study characterized semen collected from the Japanese black bear, Ursus thibetanus aponicus, to provide information on semen cryopreservation for artificial breeding. Preliminary studies using a beagle dog as the model species showed that sperm concentration and total sperm count were lower in semen collected by electroejaculation than in semen collected by digital manipulation, but that sperm motility, viability and morphology were similar. Characterization of semen obtained from Japanese black bears by electroejaculation under general anesthesia revealed that semen volume and total number of spermatozoa collected were lower; but that sperm concentration, motility, viability and morphology were equivalent to those reported in other ursids. When semen was collected via a catheter inserted into the urethra during the stimulation for ejaculation, the sperm concentration, total sperm count and motility were relatively higher than when semen was collected directly in a test tube. Specific normal semen characteristics (mean +/- SEM) were pH, 7.6 +/- 0.0; volume, 0.212 +/- 0.038 mL; sperm concentration, 361 +/- 100 x 10(6)/mL; total sperm count, 84.0 +/- 32.2 x 106; +++ motility, 30 +/- 5%; motility, 77 +/- 3%; viability 77 +/- 2%; and abnormal morphology, 11+/- 2%. These results suggest that semen can be collected from Japanese black bears by electroejaculation.     

Artificial insemination in canids: a useful tool in breeding and conservation

Artificial insemination (AI) and semen freezing have become services available to dog owners worldwide, and the demand for services to freeze semen is increasing. In other canids such as the fox, the fur industry utilizes fresh or frozen semen to artificially inseminate vixens to produce pelts. Clearly, AI facilitates the use of a male to sire several females by diluting the ejaculate, increases breeding hygiene, and allows crossing between species with slightly different breeding seasons. The African wild dog (Lycaon pictus) is currently considered by the World Conservation Union (IUCN) as one of most endangered canids. In captive populations of African wild dogs, semen has been frozen with encouraging results, using a standard cryopreservation protocol for domestic dogs, but successful AI has not been reported. In wolves, there is one report regarding the live birth of an offspring after intravaginal AI of a deslorelin-induced estrous female. In 2005, three Mexican gray wolf females were artificially bred by intrauterine insemination with freshly collected semen from unrelated males, and all females whelped. Artificial insemination may be vaginal, intrauterine or intratubal, and the semen may be fresh, fresh and chilled (diluted), or frozen-thawed, and the source of semen may be epididymal or ejaculated. In the domestic dog, the results are good to excellent for AI with all three types of processed semen when the source is ejaculated semen, whereas epididymal sperm still yields poorer results. Species differences in female physiology, as well as differences in the cryotolerance of the sperm from various canid species, warrant further research and development.     

Cryopreservation of canine semen using a coconut water extender with egg yolk and three different glycerol concentrations

Semen was collected by digital manipulation from six adult dogs. The second fraction of the ejaculate was used in this study. The semen was assessed by macroscopic and microscopic criteria 1 min after collection, first dilution, cooling, glycerol addition and thawing. Experiments were conducted to compare the effect of three different concentrations of glycerol in coconut water extender. The freezing method employed was that one described for caprine semen with slight changes. Semen was thawed at 37 degrees C for 1 min. Spermatozoal motility after thawing was 49.2+/-26, 44.2+/-18.3 and 35.8+/-26.8% for groups with 4, 6 and 8% glycerol, respectively. The vigor after thawing was 2.6+/-1.1, 2.7+/-1.0 and 2.1+/-1.2 for these groups, respectively. There was no difference among groups in motility and vigor. However, a smaller percentage of total and secondary abnormalities was observed using 6% glycerol in coconut water extender. In conclusion, the three glycerol concentrations (4, 6 or 8%) can be used successfully in cryopreservation of canine semen using a coconut water extender.