Unusual sex chromosomal DSD in a domestic Shorthair cat with a 37,X/38,XY mosaic karyotype.

BACKGROUND: Sex chromosome abnormalities associated with disorders of sexual development (DSD) are rarely described in cats, mainly due to the lack of chromossome studies that precisely reveal the condition. Genetic approaches are therefore required in order to detect sex chromossomes abnormalities as variations in the number and structure of chromosomes, or the presence of a second cell line as mosaicim or chimerism. CASE PRESENTATION: A male Shorthair cryptorchid cat was presented with clinical signs of anorexia, tenesmus and hyperthermia. Ultrasonography revealed a fluid-filled structure, with approximately 1 cm in diameter, adjacent to the descending colon. Computed tomography evidenced a tubular structure, ventral to the descending colon and caudal to the bladder, which extended cranially, through two branches. Histopathological evaluation confirmed the presence of two atrophic uterine horns and one hypoplastic testicle with epididymis at the end of one of the uterine horns. The end of the other uterine horn was attached to a structure composed by a mass of adipocytes. Cytogenetic analysis revealed a mosaic 37,X/38,XY karyotype. The two cell lines were found in 15% and 85% of the lymphocytes, respectively. Genetic analysis confirmed the presence of SRY and ZFY genes in blood and hair bulbs, and revealed a marked reduction in SRY expression in the testicle. Additionally, this case presented exceptionally rare features, such as a Leydig' cell tumour and a chronic endometritis in both uterine horns. CONCLUSIONS: Complete imaging workup, cytogenetic analysis and SRY gene expression should be systematically realized, in order to properly classify disorders of sexual development (DSD) in cats.

Influence of two collection frequency intervals on sperm quality of standard and miniature bull Terriers during short breeding periods: A clinical field study.

Background and Aims: The quality of canine sperm can be influenced by many factors, such as breed, body weight, age, ejaculatory frequency, nutrition, and environment. In the UK, it is common practice for standard Bull Terriers (SBT) and miniature Bull Terriers (MBT) to require male donors during a short breeding period. The aim of this study was to evaluate the effect of semen collection frequency on ejaculate volume and nine sperm parameters in SBT and MBT males, considering age and body condition score (BCS). Materials and Methods: Ejaculates from six adult SBTs and four MBTs were collected 5 times at two consecutive intervals (Time Series [TS]1, 24 h vs. TS2, 48 h), 1 week apart. Ejaculate volume, concentration, total output, viability (live sperm), subjective total motility, vigor, and total morphological defects, including head, midpiece, and tail defects of sperm, were evaluated. A multivariable mixed linear model for repeated measures was used to analyze the effects of semen collection frequency, age, breed, and BCS on ejaculate volume and sperm parameters. Results: Semen collection frequency, age, and, to a lesser extent, breed, and BCS significantly affected sperm parameters. Semen collection frequency affected all sperm parameters (p < 0.05) but not ejaculate volume (p > 0.05). Total sperm output, sperm vigor, total motility, and tail defects decreased (p < 0.05) at the end of TS1. However, sperm parameters remained relatively constant (p > 0.05) in TS2 between semen collection sessions. Overall, poorer sperm parameters were observed in older dogs (aged 5-8 years) than in younger dogs (aged 4 years). MBT produced less (p < 0.001) ejaculate volume (3.2 ± 0.2 mL vs. 4.3 ± 0.2 mL: Least Squares Mean ± Standard Error of Mean), lower total sperm output (221.8 ± 19.2 × 106 vs. 348.6 ± 19.2 × 106) and lower total morphological defects (25.0 ± 1.1% vs. 31.3 ± 0.9%), and a higher percentage of live sperm (77.0 ± 1.4% vs. 71.7 ± 1.1%) than SBT. In addition, a BCS of 4 positively influenced (p < 0.05) viability, vigor, and total sperm motility. Conclusion: Despite differences in age, breed, and BCS, better sperm parameter values were observed in all semen collection sessions. However, intensive semen collection (TS1) appears to be less effective in maintaining good sperm quality. For breeding or artificial insemination purposes, a 48-h interval between collection sessions is recommended for both breeds. The results of this study could be used to further optimize assisted reproductive technologies in both breeds.

Simultaneous Torsion of a Uterine Horn and Spleen in a Pregnant Two-Year-Old Crossbreed Bitch: A Case Report.

A pregnant 2-year-old mixed-breed dog was admitted with a 2-day history of lethargy, anorexia and painful abdominal distension. Clinical manifestations were unspecific and mainly suggested hypovolemic shock. Physical examination, ultrasonographic evaluation and radiographs confirmed mid-late pregnancy and evident signs of fetal death and peritoneal effusion. An exploratory laparotomy was immediately initiated which revealed torsion of the right gravid uterine horn over the left one and, simultaneously, the unexpected presence of splenic torsion and rupture, with a stretched omentum covering and exerting tension on the spleen's cranial pole. Histopathology revealed hemorrhagic infarction of the uterus and spleen. The animal recovered uneventfully and was later submitted to a preventive gastropexy.

The Comet assay for detection of DNA damage in canine sperm.

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno-, terato- or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A-median: 31.55%, interquartile range (IQR): 30.18-38.01; group B-median: 0.90%, IQR: 0.65-1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = -.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non-normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.

Interleukin-6 gene -174G>C and -636G>C promoter polymorphisms and prostate cancer risk.

Prostate cancer (PCa) is one of the most commonly diagnosed internal malignancies affecting men. Due to the important roles of IL-6 in different physiological and pathophysiological processes, IL-6 polymorphisms may modulate PCa risk. IL-6 -174 G>C (rs 1800795, also designated -236 G>C) and -636 G>C (rs 1800796, also designated -572 G>C) promoter polymorphisms have been implicated in PCa susceptibility, albeit still controversial. A literature search using PubMed and Highwire databases was conducted, resulting in eight case-control studies concerning the IL-6 -174 G>C polymorphism (11,613 PCa cases and 13,992 controls) and four case-control publications regarding the IL-6 -636 G>C polymorphism (1,941 PCa cases and 3,357 controls). In order to derive a more precise estimation, a meta-analysis based upon these selected case-control studies was performed. There was no significant association between IL-6 -174 G>C polymorphism and PCa increased risk. Nevertheless, the presence of allele C and the CC genotype were statistically significantly associated with decreased PCa risk in the overall analysis for IL-6 -636 G>C polymorphism. Additional studies in larger samples and analyses of functional repercussions of these SNPs in prostate tumor cells are necessary to validate these findings.

Priapism secondary to perineal abscess in a dog - a case report.

A 7-year-old intact male Boxer was referred to our services at the Veterinary Teaching Hospital of the University of Trás-os-Montes and Alto Douro, suffering from a persistently erect penis (including the bulbus glandis) that had been exposed for several days. Radiographic and ultrasonographic examinations detected a 5.0 x 3.5 cm mass located dorso-laterally to the urinary bladder. The microbial culture of the mass revealed Staphylococcus spp. At that time, we suspected the involvement of an abscess in the origin of the priapism. Medical and surgical treatments were promptly instituted, which allowed for penile withdrawal into the prepuce; however, the resolution of the penile erection was not accomplished in the following days and penile amputation was required. Histological evaluation of the excised penis revealed extensive infarction of the erectile tissue of the pars longa and bulbus glandis, and also of the blood vessels of the penis. Following penile amputation and antimicrobial therapy, the animal fully recovered. Ultimately, the animal died as a consequence of gastric torsion. At necropsy, some lesions compatible with a previous perforation of the intestinal wall were recorded. The data gathered from the anamnesis, the physical and imaging examinations, along with the post-mortem findings, allowed us to conclude that in this clinical case the primary cause of priapism was a perineal abscess due to bowel perforation.

Effects of taurine and hypotaurine supplementation and ionophore concentrations on post-thaw acrosome reaction of dog spermatozoa.

The aim of this work was to study of the effect of the amino acids (AA) taurine (T) and hypotaurine (H) and of different calcium ionophore concentrations on the ability of capacitated frozen-thawed dog sperm to undergo the acrosome reaction (AR). Fifteen ejaculates grouped into five pools were used. Sperm was frozen at a concentration of 80x10(6)sperm cells/mL in the Uppsala Equex extender (UE) supplemented with 25, 50 and 75mM of either AA. The UE extender without T or H was used as control. After thawing, sperm was capacitated with Canine Capacitation Medium for 20min. Sperm was then challenged with calcium ionophore A23178 at 0, 2.5 and 10microM concentration and evaluated for integrity of plasma and acrosome membranes after 5, 15 and 30min of incubation, utilizing PI/Fitc-PNA fluorescent staining and flow cytometry. Sperm cryopreserved in UE supplemented with 50mM T (UE 50T) had higher AR rates than sperm cryopreserved with UE 75T, UE 25H and UE 50H, but AR rates were similar to semen frozen with the control extender. Challenges with 2.5 and 10microM/L of calcium ionophore increased AR in frozen-thawed sperm incubated for 5, 15 and 30min. The combination of calcium ionophore concentration and incubation time resulting in the highest AR rate was 10microM and 15min.

Differences in preservation of canine chilled semen using different transport containers.

In the present study, the effect of three different containers in the preservation of dog chilled semen, during 24, 48 and 72h was evaluated. Weekly sperm pools of different dogs were obtained, during 10 consecutive weeks. Semen samples were diluted in egg-yolk-Tris-fructose extender and stored in a Styrofoam box, a common Thermos flask and an Equitainer. Progressive motility, morphology and sperm membrane integrity were examined in semen aliquots taken daily from each container during the 3 days of storage. Additionally, integrity of the acrosome and sperm plasma membranes, determined by PI/Fitc-PSA staining was assessed at 48 and 72h of storage. At 24h no differences were observed between the three containers for the evaluated parameters. At 48h samples kept in the Equitainer presented a higher progressive motility than samples kept in the Thermos. At 72h, progressive motility was higher in the Equitainer than in the other two containers. Only samples kept in the Equitainer maintained similar levels of progressive motility between 24 and 72h. Membrane integrity assessed by eosin-nigrosin deteriorated over the 72h period, whereas functional membrane integrity determined by the hypoosmotic swelling test was independently affected by type of container (the Equitainer) kept a higher percentage of sperm cells with intact membrane) and time of storage (a decrease of membrane integrity between 24 to 72h). Staining with PI-Fitc-PSA allowed the detection of differences between containers but not between the two studied storage periods (48 and 72h). The results indicated that the use of the Equitainer is preferable when transporting chilled dog semen for more than 48h.

Incorporation of taurine and hypotaurine did not improve the efficiency of the Uppsala Equex II extender for dog semen freezing.

The working hypothesis of the present study was that supplementation of the Uppsala Equex II (UE) extender with the amino acid (AA), taurine (T) and hypotaurine (H) would improve dog sperm post-thaw quality, as previously seen for ram and bull semen, respectively. Five pools from 15 ejaculates of 15 dogs were used. Each AA was added to the UE extender at a concentration of 25, 50 and 7 5mM. Amino acid-free extender was used as a control. The following post-thaw parameters were evaluated: sperm motility by light microscopy and by CASA evaluation, longevity, viability (eosin-nigrosin staining), and flow cytometry (FC) was used to assess acrosome integrity and mitochondrial activity after PI/Fitc-PSA and PI-Rhodamine staining, respectively. Post-thaw sperm motility and velocity did not differ among extenders. Amplitude of lateral head displacement was lower for sperm frozen in the 25 mM H-supplemented extender. Semen frozen in the extender with 50 mM of T resulted in higher number of live sperm with damaged acrosomes after thawing. Higher numbers of live sperm with minimal mitochondrial activity were obtained for samples frozen with 25 and 50 mM T-supplemented extenders. Semen frozen in the control and 50 mM T-supplemented extenders had the highest number of live (eosin-nigrosin stain negative) sperm immediately post-thawing. We concluded that supplementation of the Uppsala extender with T or H did not improve sperm post-thaw mitochondrial activity or semen motility and viability.