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Hematuria, or preputial hemorrhagic discharge, is an extremely common clinical sign; it can be associated with a wide range of diseases, including, even if only rarely, penile foreign bodies. The aim of this retrospective study was to describe the diagnosis and therapy involving migration from the preputial ostium or penile urethra of a foreign body (awn grass) embedded in the connective tissue surrounding the penis, or in deeper inguinal tissues, in dogs with hematuria or preputial hemorrhagic discharge. In the selected cases, signaling, history, clinical signs, results of laboratory exams, endoscopic and ultrasonographic evaluation, and the technique used for foreign body removal were evaluated. Dogs with hemorrhagic discharge consequent to a penile foreign body represented 2% of the entire population considered. At physical examination, the most common features were the presence of swelling of the glans and hyperemia associated with a penile fistula (4/6 dogs), and pain during penile exteriorization (3/6 dogs). Laboratory results showed mild neutrophilic leukocytosis in 2/3 dogs and reticulocytosis in 1/3 dogs. Endoscopy, performed in 2/6 dogs, did not reveal any alterations associated with passage of the foreign body. Ultrasonography was useful in reaching a definitive diagnosis, identifying the position of the grass awn in 6/6 cases and permitting its removal in all dogs using an ultrasound-guided technique. This case report suggested that penile foreign bodies are a rare, but possible, cause of hematuria or hemorrhagic discharge in male dogs, and that ultrasonography is a useful technique in the making of a differential diagnosis and removal of foreign bodies.

Placenta is essential for the development of the fetus, and its impaired function can lead to a negative outcome (i.e., neonatal mortality). In dogs, investigations on placenta histology and neonatal outcome in healthy bitches are lacking, and a contribution is provided in this study to emphasize the use of placenta histology in practice. Fifty-one placentas from 11 litters were collected during cesarean section, classified according to the litter size (large (L) or small (S)) and the outcome, this latter as healthy (Group 1) or dead within 7 days (Group 2). The placenta/puppy weight ratio (PPR) was calculated, and specimens were formalin-fixed and paraffin-wax embedded, and on the resulting histological slides, capillary density (CD) was quantified. Among necrosis, calcification, and intravascular leucocytes, only the presence of multifocal-confluent necrosis (significantly more frequent in Group 2) was associated with a higher risk of death within 7 days (odds ratio = 30.7). Mixed logistic regression ruled out the effect on death both of a bitch and cesarean type (programmed vs. emergency). PPR and CD values were associated with litter size, large litters had lower PPR (p &lt, 0.01) and higher CD (p &lt, 0.05) than small litters. The relationship between PPR and CD was negative and significant (p &lt, 0.01). Necrosis was a frequent finding in canine placentas, but only when multifocal-confluent was it associated with a poor outcome. The litter size influenced PPR (lower in L) and CD (higher in L), and this is likely due to the plasticity of placenta adaptation.

High dose medetomidine 0.13 mg/kg can be used for semen collection in cats with variable results in terms of quantity and quality. Therefore, a variation in terms of distribution and elimination among patients has been hypothesised. The aim of the study was to characterise the pharmacokinetics of medetomidine (0.13 mg/kg) administered intramuscularly (IM) in healthy male cats. Eighteen male cats undergoing castration were included, and medetomidine (0.13 mg/kg) was administered IM. Venous blood samples were collected at 20, 30, 40, 50, 60, 75 and 90 minutes after medetomidine administration. Before orchiectomy, at T20, sperm collection was attempted. Plasma medetomidine concentrations were determined by liquid chromatography/mass spectrometry analysis. Semen collection was successful in 15/18 cats. The medetomidine plasma concentration following the IM administration of a bolus was best described using a non-compartment model. Time of maximum concentration was observed at 40 minutes (range 20−90); maximum concentration was 32.8 ng/mL (range 26.8−51.2). The median apparent clearance was 11.9 mL/kg/minute (range 0.7−43.8). In conclusion, medetomidine administered IM at 0.13 mg/kg reached its peak plasma concentration slowly and with variability among patients. In addition, it was characterised by low total body clearance probably due to the cardiovascular alterations associated with medetomidine administration.

Egg yolk (EY) is conventionally used to reduce sperm cryodamage, however, there has not be evaluation of whether there is a dose-dependent effect with inclusion of EY in semen extender. To enhance the knowledge about the protective effect of EY during cryopreservation of dog semen, a specific study was designed to evaluate the dose-dependent protection of the EY against osmotic and cryogenic damage of dog sperm. In the first experiment, sperm stored in an extender that contained graded EY concentrations (0 %, 5 %, 10 %, and 20 %) were diluted with hypo- or hyper-osmotic solutions (final osmolality of 75, 150, 300, 500, 1000 mOsm/kg). Results from sperm kinetic, membrane integrity (MI), mitochondrial activity, and normal morphology evaluations indicated osmotic stress has especially marked effects on the kinetic capacity of spermatozoa, however, there were no direct effects on mitochondrial activity. In both hypo- and hyper-osmotic conditions, EY had a protective effect regardless of concentration. In the second experiment, semen samples were diluted in extenders at increasing EY concentrations (0 %, 5 %, 10 %, and 20 %) and cryopreserved. Effects on sperm kinetics, membrane and acrosome integrity and mitochondrial membrane potential indicated there was improved sperm viability after thawing when the EY concentration was 5 % and 10 %, and lesser viability when it was 20 %. These results indicate, for the first time, that EY reduces osmotic and cryogenic damage when used at 5 % or 10 % concentrations, and that these concentrations can be used to protect dog spermatozoa more effectively than the conventionally used concentration (20 %).

Dog sperm cryopreservation is gaining importance both in breeding dogs for commercial purposes and for pet animals. Anyway, cryopreservation of mammalian spermatozoa, including dog ones, induces some negative effect on sperm fertility, leading to a lower use of this technique and limiting its widespread use. Therefore, studies to improve the quality of canine semen after cryopreservation could have a relevant impact on both the scientific advancement and the clinical practice. The aim of the present work was to investigate the putative ameliorative effect of Epigallochatechin-3-gallate (EGCG) addition to post thawing medium on dog sperm motility, mitochondrial activity, acrosome integrity and on zona-binding ability (zona binding assay). Spermatozoa were thawed in Tris-fructose-citrate medium supplemented with EGCG (0, 25 and 50 μM) and sperm motility, mitochondrial activity and acrosome integrity were assayed at 0.5, 1.5, 3 and 6 h after post thawing incubation at 37 °C. An aliquot of semen from each treatment group after 1.5 h post thawing incubation was washed and used to perform heterologous (using porcine oocytes) or homologous zona binding assay. The results obtained showed that no significant effect is exerted by EGCG on sperm parameters analysed neither at 0.5, 1.5, 3 or 6 h after thawing excepting for the reduction of the percentage of live cells with active mitochondria at the higher dose at 6 h; furthermore, both homologous or heterologous zona binding ability, was not influenced by EGCG. In conclusion, EGCG supplementation to thawing medium does not improve dog sperm quality or zona binding capacity.

Objectives This study aimed to assess non-invasively the cardiovascular effects of high-dose medetomidine on healthy male cats undergoing semen collection. Methods Haemodynamic evaluations were assessed on the basis of clinical examination, systolic arterial pressure (SAP) and transthoracic echocardiographic examination. Eight client owned, male domestic shorthair cats were sedated with a bolus of medetomidine intramuscularly (IM; 0.13 mg/kg), and semen collection was performed. A second transthoracic echocardiographic examination and SAP measurement were carried out 15 mins after sedation. At the end of the examination, the patients received a bolus of atipamezole (0.3 mg/kg) IM. Results The cats were deeply sedated, relaxed and laterally recumbent during the entire procedure. No rhythm abnormalities were observed during the examinations and no significant increase in SAP was recorded. Heart rate dropped from 200 ± 33 to 92 ± 13.1 beats per min after sedation. There was a significant increase in left ventricular dimensions and the left atrial area. The parameters of left ventricular systolic function were reduced, as were systemic and pulmonary cardiac outputs. Peak diastolic wave velocities were significantly reduced, while isovolumic contraction and relaxation time of the left ventricle were prolonged. Aortic valve insufficiency was recorded for all cats, while mitral valve insufficiency was noted in five cats. None of the subjects developed systolic anterior motion of the mitral valve. Conclusions and relevance The protocol allowed us to collect good semen samples in healthy cats. However, high-dose medetomidine induces significant haemodynamic effects on the feline heart, mainly due to a reduced heart rate, an increased cardiac preload and impaired systolic function. The animals recovered from the anaesthesia, after antagonist administration, without showing any clinically relevant consequences.

The present study consists of two distinct parts, experiment 1 and experiment 2. In experiment 1, 13 anestrous queens were treated with a 4.7-mg deslorelin subcutaneous implant to assess its effectiveness in inducing estrus in the domestic cat. Deslorelin is currently used for the reversible suppression of ovarian and testicular activity in dogs and cats and for estrus induction in the bitch. Estrus induction is also reported in the queen but never reported with a targeted study. All the queens showed a positive response to the induction protocol, and estrus was detected within an average of 5.0 ± 2.2 days after the implant placement in 13 out of 13 subjects (100%). Seven of 13 queens exhibited behavioral manifestations of estrus, and the mean number of follicles detected at ultrasound examination was 4.8 ± 1.6 per subject. In experiment 2, three of the queens previously treated with deslorelin for estrus induction were submitted to artificial insemination through endoscopic transcervical catheterization, a new nonsurgical technique for intrauterine sperm deposition. All of them (100%) were pregnant after insemination and they gave birth to healthy litters. The study, as a whole, proves the effectiveness of the 4.7-mg deslorelin subcutaneous implants in inducing estrus in the domestic cat and is, to our knowledge, the first study assessing fertility of the induced estruses. Moreover, it shows the effectiveness of endoscopic transcervical catheterization for artificial insemination in the queen.

This study was designed for the identification of different sperm kinetic subpopulations in feline semen using artificial neural networks (ANNs) and for the evaluation of the effect of ejaculation on motility patterns of these subpopulations. Seven tomcats presented for routine orchiectomy were electroejaculated, and after 5 days, orchiectomized and epididymal tail sperms were collected. Sperm motility characteristics were evaluated using a computer-assisted sperm analyzer that provided individual kinetic characteristics of each spermatozoon. A total of 23 400 spermatozoa for electroejaculated and 9200 for epididymal tail samples were evaluated using a multivariate approach, comprising principal component analysis and ANN classification. The multivariate approach allowed the identification and characterization of three different and well-defined sperm subpopulations. There were significant differences before (epididymal tail spermatozoa) and after (electroejaculated sperm) ejaculation in sperm kinetic subpopulation characteristics. In both epididymal and ejaculated samples, the majority of subpopulation was characterized by high velocity and progressiveness; however, the electroejaculated samples showed significantly higher values, suggesting that the microenvironment of the epididymal tail could affect the sperm motility or, alternatively, seminal plasma could increase the kinetic characteristics of the spermatozoa, indicating that only after ejaculation, the spermatozoa express their motility potential. Nevertheless, further studies are required to clarify the functional significance of each kinetic subpopulation.

This paper aimed to evaluate cat seminal plasma protein profile (with SDS-page) and determine differences in seminal plasma composition from ejaculates obtained using urethral catheterization after pharmacological induction (UrCaPI) and electroejaculation (EE). In addition, this study evaluates whether the recovery method affected seminal plasma protein and zinc concentrations. A single ejaculation was collected from 17 mixed-breed cats by EE (5/21) or UrCaPI (12/21), while 4/21 cats underwent four sperm collections once every four days using EE and UrCaPI techniques alternately. The semen parameters evaluated were: volume, percentage of motility and progressive motility, morphology, and sperm concentration. After centrifugation, the seminal plasma obtained was stored at -80 °C and later used to measure protein and zinc concentrations, and to determine protein profile by SDS-polyacrylamide gel electrophoresis (PAGE). The results obtained indicate that cat seminal plasma protein profile is characterized by many protein bands (>30) with a molecular weight ranging from 3.5 to 200 kDa, and that the recovery method influences the seminal plasma protein profile: EE is related to the absence of two proteins (P55 and P14), and alters three protein bands (P200, P80, P28). The collection technique also affected zinc concentration (mg/dL) and protein concentration (g/dL) which were significantly higher (P < 0.01) in samples collected by UrCaPI; on the contrary the total Zn and protein amount/ejaculate were not significantly different in samples collected by both technique (P < 0.05).

Contents Semen collection and AI in the cat are still not routine procedures. The correlation between semen quality and fertility under natural conditions is a relatively unknown field in the cat. In the present study, functional in vitro tests, such as the ability to bind and penetrate the zona pellucida or to fertilize in vitro, were used to determine fertilizing ability of sperm cryopreserved with a practical and efficient freezing protocol previously developed in our laboratory. Semen was collected by electroejaculation, evaluated for motility and diluted with Tris–glucose–citrate egg-yolk extender supplemented with Equex STM paste (0.5% v/v). After equilibration and loading into 0.25 ml straws, semen was frozen at 3.85°C/min. Frozen-thawed semen was co-cultured with in vitro matured cat oocytes. Penetration rate was recorded 30 h after in vitro fertilization and cleaved zygotes were cultured in vitro until day 7. A correlation was found between sperm motility index (SMI) after thawing and semen fertilizing ability (p

The objective of this study was to evaluate the influence of different concentrations of epidermal growth factor (EGF) on in vitro maturation of domestic cat oocytes. A total of 444 cat oocytes were matured in MSOF (maturation synthetic oviductal fluid) in the presence of varying EGF concentrations: (1) MSOF (control); (2) MSOF+10 ng/mL EGF (EGF10); (3) MSOF+25 ng/mL EGF (EGF25); and (4) MSOF+50 ng/mL EGF (EGF50). After IVM, oocytes were in vitro fertilized to verify the effect of adding EGF on cytoplasmic maturation. Cleavage rate was recorded and noncleaving oocytes were stained with Hoechst 33258 and examined to determine nuclear maturation rate. Cleaved zygotes were cultured in vitro and embryo stages were evaluated on days 6 and 7. There was no difference among groups in the total number of oocytes reaching the metaphase II (MII) stage (P0.05). The EGF25 group had the highest (P0.01) blastocyst yield (37.5%) and developmental competence (60.9%). Cleavage rate and resulting morulae and blastocysts on day 6 for EGF25 group were higher (P0.01) than control and EGF50 groups. Although EGF did not significantly enhance nuclear maturation rate, it had a dose-related positive effect on cytoplasmic maturation, since the oocyte's ability to cleave and reach the blastocyst stage was improved at 25 ng/mL, with intermediate improvement at 10 ng/mL, but 50 ng/mL had no significant benefit. In conclusion, the addition of EGF to the maturation medium enhanced cytoplasmic maturation of cat oocytes in vitro.

A thorough knowledge of the genital tract anatomy and vaginal and cervical modifications during the phases of estrus cycle of the female cat is necessary to enhance success of assisted feline reproductive technologies or for other diagnostic or therapeutic purposes. There are few studies of the gross anatomy of the genital tract in the queen. The vestibule is large in comparison to the vagina; the latter has a narrow lumen, which is further constricted by the prominent dorsal median fold. The cervix is oriented obliquely between the uterine body and vagina. Vaginal and cervical changes in association with the estrous cycle have been reported in some species and two recent studies investigated this aspect in the cat. This paper reviews the relevant literature, with particular emphasis on recent studies.

In a previous study we observed that it is possible to reach the cervix in all queens with a 1 mm diameter probe only. So, we developed both a new technique and a catheter (1 mm diameter) to allow transcervical insemination [Zambelli and Castagnetti 2001]. The aims of this study were to investigate vaginal and cervical anatomic modifications during the various stages of the oestrus cycle and to test the previously described technique of transcervical catheterization during the various stages of the oestrus cycle. In experiment 1, silicon impression moulds were obtained from the reproductive tracts of 21 queens' cadavers and vaginal and cervical measures were taken. The results showed that there are some significant anatomic modifications during the various stages of the oestrus cycle in vaginal and cervical anatomy, principally related to the dorsal medial fold increase induced by the follicular phase. In experiment 2, transcervical catheterization was attempted in 95 queens at various stages of oestrus cycle both during reproductive and non-reproductive season. After catheterization, methylene blue solution was injected through the cervical catheter. Successful catheterization was assessed during surgery, when colour was observed in the uterine horns. It was possible to perform transcervical catheterization during non-reproductive season in 16 of 20 anoestrus queens and in 12 of 15 induced oestrus queens; during reproductive season in nine of 21 interoestrus queens, in eight of 13 dioestrus/pregnancy queens, in four of 18 oestrus queens and in seven of eight queens in first oestrus during lactation.