Case of Anorchia in a Mixed-Breed Dog

Anorchia is a rare and often poorly understood disorder of sexual development. In the present case report, we used a multidisciplinary approach to diagnose a case of anorchia in a 30-month old dog. The diagnostic process began with gathering the dog medical history followed by a clinical visit with the patient, which included a general wellness examination as well as an examination of the genital system. As suggested in the relevant literature, the dog underwent an ultrasound and then computed tomography (CT) of the genital system, which confirmed the diagnosis of testicular agenesis. Genetic testing confirmed the male XY karyotype. Hormone testing also supported the diagnosis: testosterone and anti-M€ullerian hormone levels were below their reference ranges, and luteinizing hormone (LH) was above 1 ng/mL. Following the diagnostic procedures suggested in the relevant literature, the present study confirms anorchia in the dog and describes a case of testicular agenesis in the canine species

Evaluation of the analgesic effect of the Fentanyl patch during ovariectomy and the postoperative period in bitch.

For several years now, routine surgical procedures such as ovariectomy are performed as outpatient surgery. Post-operative care turns out to be much more comfortable for the patient than hospitalization, particularly at home in reducing the risk of nosocomial infections. However, this approach rises the problem of the administration of analgesic drugs by the owners at home. The potent opioid fentanyl is market in the form of a patch (DURAGESIC®, Jannsen), which fentanyl is gradually released for approximately 72 hours after application (1). The aim of this study was to evaluate the convenience and the analgesic efficacy of this patch, in comparison to other analgesic protocols usually utilized in ovariectomized bitches. This study involved 20 privately-owned bitches, ovariectomized at the Veterinary Hospital of the Department of Veterinary Medicine (Bari). Only bitches housed without other domestic animals were selected, to allow the owners to perform a strict control in the postoperative period. The animals (healthy and from 2 to 5 years of age) were randomly divided into two 2groups. In both groups after premedication with acepromazine, anesthesia was induced by propofol and maintained with isofluorane. The following parameters were monitored during the procedure in all animals: heart rate, electrocardiogram, EtCO2, pulse oximetry, blood pressure and body temperature. The analgesic protocols in the Fentanyl group and Control group were different. The Fentanyl patches were applied 12 hours before the surgery. Transdermal fentanyl patches were placed on the neck skin, which had been previously shaved and disinfected. The size of individual patch was chosen based on the patient's body weight: <10 kg = 25 mcg/h; 10-20 kg = 50 mcg/h; 20-30 kg = 75 mcg/h, and 30-40 kg = 100 mcg/h (2). The patches were affixed with adhesive tape, and left on site for 72 hours after application (3). In the control group standard analgesic protocol pre-surgically was with methadone and oral administration of analgesic drugs at home was administered (robenacoxib). In the clinic, after the dog awakened from anesthesia, the behavior and level of pain/discomfort were evaluated using the Glasgow Pain Scale. The owners filled out a questionnaire with a numerical scale (from 0 to 10) to assess their dogs behavior and the owners satisfaction, regarding the management of their animals postoperative. All the animals in the study hade an uneventful recovery from anesthesia. The owners of the bitches in Fentanyl group were more satisfied with post-operative management than were the owners of the control group (administration of drugs orally). More pain was reported in the control group, related to bitches refusing oral medication. The bitches in the Fentanyl group tolerated the Fentanyl patch well, without any side effects noted. The data obtained from this study shows that the fentanyl patch is a valid aid for effective analgesia post-ovariectomy in bitches

Bioenergetic/oxidative balance in feline morulae and blastocysts produced in vitro after temporary oocyte holding at room temperature versus cold ovary storage.

Maternal aging is associated with an increase in embryonic aneuploidy and early miscarriage, as a result of errors in chromosome segregation [1]. Failure to achieve correct alignment of the chromosomes on the spindle is an important factor contributing to mis-segregation errors in oocytes [2,3]. Since little data is available in horses, the aim of this study was to evaluate the effect of maternal aging on spindle morphology and the incidence of chromosomes misalignment. Cumulus oocyte complexes (COCs) were recovered from slaughtered mares, divided into groups depending on mare age (young, < 15 years; old, ≥15 years) and matured in vitro for 26h. After maturation and denudation, only the oocytes that had reached MII were used and these were further subdivided into Control and Nocodazole groups (total = 4 groups; n=20 per group). Oocytes in the Nocodazole groups were incubated for 10 min in medium containing 20 μM Nocodazole, to induce tubulin depolymerization, washed and then incubated for 2 hrs in maturation medium. In order to destabilize any tubulin fibers not properly attached to the kinetochores samples were subjected to cold shock, fixed and stored at 4 ̊C prior to immunofluorescent staining for DNA and alphatubulin. Spindle morphology and the incidence of chromosome misalignment were evaluated by confocal microscopy and 3D image analysis (Imaris 8.3). Spindle morphology was scored as normal (fusiform, bipolar) or abnormal (tri- or tetra-polar, severely misshapen), chromosome misalignment was scored as absent, mild (1-5 misaligned chromosomes) or severe (>5 misaligned chromosomes). Oocytes from aged mares showed higher rates of mild and severe chromosome misalignments when compared to those from young mares, both in normal condition (mild 37 vs 5%; severe 11 vs 0%) and after Nocodazole treatment (mild 42 vs 15%; severe 21 vs 0%). Moreover, oocytes from old mares were more likely to show abnormal spindle morphology both under control conditions (5 vs 0%) and after Nocodazole treatment (10 vs 0%). Although nocodazole treatment did not result in a significant increase in chromosome misalignment and spindle abnormalities, the incidence of chromosomal misalignments increased numerically in both aged and young groups (total % of misalignment without treatment 47.4 and 4.5% vs 63.2 and 15% after nocodazole treatment). We suggest that the compromised ability to form a normal meiotic spindle and correctly align the chromosomes observed in MII oocytes from aged mares might reflect an impaired function of the spindle assembly check point components, and explain the age-related reduction in oocyte developmental competence

EXPRESSION AND SUBCELLULAR LOCALIZATION OF THE M-OPIOID RECEPTOR IN EQUINE SPERMATOZOA. EVIDENCES FOR ITS FUNCTIONAL ROLE

The development of fertilizing ability in sperm cells is associated with changes in the plasma membrane. However, to date the exact nature of sequentially activated primary receptors and channels and the signal transduction pathways derived from these remains elusive. We analyzed the expression and localization of the m-opioid receptor in equine spermatozoa. A transcript corresponding to the third extracellular loop that selectively binds m agonists was amplified, sequenced and compared with the known sequences in humans, rats and cattle. The amplification product showed a high degree of nucleotide conservation. By immunofluorescence, m-opioid receptor labeling was found on the sperm head and on the tail and disappeared in the acrosomal region of acrosome-reacted sperm cells. Immunoblotting revealed two bands of 50 and 65 kDa. Effects of the opioid antagonist naloxone on motility and on viability and capacitation/acrosome reaction were investigated by computer assisted sperm analysis and Hoechst 33258/chlortetracycline (H258/CTC) staining. Progressive motility was significantly reduced after 3 h incubation in 1023M naloxone (P < 0.05), whereas it increased significantly after 5 h in 1028M naloxone (P < 0.05). Sperm velocity at 5 h was significantly reduced by the addition of 1023M naloxone (P < 0.05), but increased significantly in the presence of 1028M (P < 0.001). Curvilinear velocity and amplitude of lateral head displacement in spermatozoa incubated in the presence of naloxone were not indicative of hyperactivation. H258/CTC staining showed that 1028M naloxone significantly stimulated capacitation (P < 0.01) after 3 h. However, it had no effect on sperm cell viability and acrosomal status. Overall, this study provides the first evidence that the m-opioid receptor is expressed in equine spermatozoa and that naloxone significantly affects motility and capacitation