Contrast-Enhanced Ultrasound (CEUS) Evaluation of Canine Prostatic Hyperplasia before and after Osaterone Acetate Therapy: Preliminary Results.

The prostate is the only sexual gland of the male dog, and dihydrotestosterone (DHT) regulates its growth. In intact dogs, constant DHT stimulation results in benign prostatic hyperplasia (BPH) that can be treated with osaterone acetate (OSA). This study describes the effects of OSA treatment, detected by contrast-enhanced ultrasonography (CEUS), highlighting prostatic vascularization with a contrast agent composed of gas microbubbles. Fifteen dogs (2-8 years) of different sizes and breeds (4-30 kg) diagnosed with BPH are involved in the study. Before treatment (D0), CPSE is measured (294.05 ± 115.97 ng/mL), and a B-mode ultrasound is performed (Vratio = 2.80 ± 1.85), confirming BPH. CEUS highlights the length of the wash-in (11.93 ± 2.08 s) and wash-out (42.20 ± 6.99 s) phases of the contrast agent in the prostate and the presence of cysts and parenchymal alteration. Dogs are treated with OSA (0.5 mg/kg for 7 days) and reassessed after 21 days (D1): CPSE and prostate volume are significantly (p < 0.001) reduced. The length of the wash-in (14.73 ± 2.54 s) and wash-out (51.13 ± 6.03 s) phases are significantly (p < 0.001) increased. The results confirm the effectiveness of the treatment, particularly the reduction in prostatic perfusion, confirmed by the increase in diffusion times of the contrast. Although preliminary, these findings are promising for the use of CEUS in monitoring dogs with BPH.

A rare case of adventitious placentation (diffuse semi-placenta) in a Jersey cow.

Placental abnormalities more frequently occur during pregnancy of somatic cell clones and may lead to pregnancy loss or dystocia. Adventitious placentation, or diffuse semi-placenta, is determined by the development of areas of accessory placentation between the cotyledons due to the abnormal growth of placentomes.After a full-term pregnancy, a 3-year-old Jersey heifer was referred for dystocia which resulted in the delivery of a dead calf. The cause of dystocia was found to be foetal malposition, while the placenta was physiologically expelled after dystocia resolution.Grossly, cotyledons appeared reduced in size and number in one placental horn, while the surface of the other horn was covered with microplacentomes. Numerous villous structures without trophoblastic coating were highlighted after histopathology. The dominant sign was an inflammatory reaction. The findings were consistent with inter-cotyledonal placentitis, which led to adventitial placentation.Diffuse semi-placenta compensates for the inadequate development of placentomes and may occur as a congenital or acquired defect. The outcome depends on its severity: in the worst scenario, pregnancy may not proceed beyond midterm and may be complicated by hydrallantois. In the case under examination, the dimensions of the cotyledons (from 2 to 10 cm) allowed for the natural course of pregnancy.

First report of whole CFTR gene duplication in a healthy newborn carrying R74W and V855I variants on the same allele.

Cystic fibrosis (CF) is the most common severe autosomal recessive genetic disorder among Caucasians. The improvement of genetic techniques has allowed the identification of an increasing number of genetic variants, including large rearrangements such as duplications. We report the first case of a whole CFTR gene duplication in a healthy newborn, who had normal sweat test, also carrying R74W and V855I variants on the same allele. Familial segregation analysis and the observed frequencies of all the CFTR gene variants, revealed that R74W and V855I were probably both present in a cis arrangement on the allele also containing the duplication (i.e., in a double complex allele). Since R74W is a "variant of varying clinical consequence" its arrangement in trans with one pathogenic variant may not be sufficient to cause a classic CF disease phenotype. Moreover, its duplication could even be an advantage that could compensate for the effect of the alteration.

Evaluation of Testicular Volume and Correlation with Sperm Production in Martina Franca Donkeys: A Parameter to Consider When Approving Breeding Jacks.

Good sperm production is a notable requirement for subjects intended for reproduction, particularly in endangered species, and it has been demonstrated that in horse stallions, this is correlated to testicular volume. The present study, which involved Martina Franca jacks, aimed to determine whether, also in this endangered breed, there is a correlation between the total sperm number (TSN) and testicular volume. Testes were measured with both ultrasound and a caliper. Testicular volume was calculated using two different formulas: one representing the volume of an ellipsoid and one developed to describe round-shaped testicles. The average sperm concentration was 380.14 ± 254.58 × 106/mL, while the average TSN was 16.34 ± 7.76 × 109. Our findings evidenced a significant correlation (r > 0.75; p < 0.05) only between sperm production and the volume calculated with the formula V (cm3) = 33.57 × H - 56.57 for round-shaped testes. Moreover, significance was evidenced only for data obtained with ultrasound (VTs-us 315.03 ± 25.83 cm3) but not with caliper. In conclusion, testicular volume can be suggestive of good fertility in Martina Franca jacks; thus, this parameter could be considered when selecting breeding animals.

A Rare Case of Concurrent 2q34q36 Duplication and 2q37 Deletion in a Neonate with Syndromic Features.

Large-scale genomic structural variations can have significant clinical implications, depending on the specific altered genomic region. Briefly, 2q37 microdeletion syndrome is a prevalent subtelomeric deletion disorder characterized by variable-sized deletions. Affected patients exhibit a wide range of clinical manifestations, including short stature, facial dysmorphism, and features of autism spectrum disorder, among others. Conversely, isolated duplications of proximal chromosome 2q are rare and lack a distinct phenotype. In this report, we provide an extensive molecular analysis of a 15-day-old newborn referred for syndromic features. Our analysis reveals an 8.5 Mb microdeletion at 2q37.1, which extends to the telomere, in conjunction with an 8.6 Mb interstitial microduplication at 2q34q36.1. Our findings underscore the prominence of 2q37 terminal deletions as commonly reported genomic anomalies. We compare our patient's phenotype with previously reported cases in the literature to contribute to a more refined classification of 2q37 microdeletion syndrome and assess the potential impact of 2q34q36.1 microduplication. We also investigate multiple hypotheses to clarify the genetic mechanisms responsible for the observed genomic rearrangement.

Apgar Score for Newborn Dog Viability Assessment: Differences between English and French Bulldogs Born via Cesarean Section.

Even if largely used in canine neonatology, some questions about the Apgar Score (AS) arose. Notably, considering the breed-oriented modifications of the score are needed, slight changes of the score were reported for English (EBs) and French bulldogs (FBs). The present study aimed to evaluate the possible differences between neonatal viability of the two breeds assessed by AS in puppies born via cesarean section. The results obtained from 99 puppies born alive and without malformations (EB = 47, FB = 52) showed significant (p < 0.001) differences in the median AS (seven in EB vs. eight in FB), with Grimace (p < 0.05 for sub-score 0 and p < 0.001 for sub-score 2) and Attitude (p < 0.05) being differently sub-scored between the two breeds. In the 89 alive puppies at 7 days of age, the same difference in the median AS was observed (p < 0.001), and only Grimace was differently sub-scored between the two breeds (p < 0.05 for sub-score 0 and p < 0.01 for sub-score 2). These results suggest that low Grimace could be an intrinsic characteristic of EB newborns, but it could negatively affect the neonatal adaptation process of EBs, providing an indication for focused neonatal assistance. Neonatal mortality at 7 days of age was higher in EBs than in FBs (17 vs. 3.8%, respectively), which deserves further investigations. The study provides further evidence that breed-oriented ASs are needed for a better evaluation and assistance of purebred puppies at birth.

Canine prostatic serum esterase and strain and 2D-shear wave sonoelastography for evaluation of normal prostate in dogs: Preliminary results.

Canine prostatic serum esterase (CPSE) is considered a useful tool to identify prostate disorders in dogs, with increasing interest in ultrasound (US)-based sonoelastography to non-invasively detect prostate disorders. Since no report is available about a possible correlation between these diagnostic tools, we aimed to investigate a possible correlation between strain elastography (SE) and 2D-shear wave elastography (SWE) and CPSE. Twenty-one dogs were included and, on each animal, CPSE was evaluated followed by a complete US examination and SE and 2D-SWE application. Healthy dogs were identified based on the CPSE results. All the dogs included were characterized by normal CPSE values (<52.3 ng/mL) and normal US prostate appearance. The prostate was characterized by intermediate stiffness with SE (pattern III - 84.7% for the left lobe and 79.27% for the right lobe) and softer than the abdominal wall (SR 0.6 for the left lobe and 0.56 for the right lobe), with low values for both m/s and kilopascals (kPa) for 2D-SWE, pointing that the healthy tissue is not hard. 2D-SWE results were, respectively, 13.51 ± 5.55 kPa and 2.31 ± 0.42 m/s for the left lobe and 18.05 ± 6.47 kPa and 2.39 ± 0.43 m/s for the right lobe. The significant difference between the right and left measurements expressed with kPa, not evidenced with m/s, can be considered indicative of m/s as the most reliable measurement to be considered regarding the prostate parenchyma. Even though no linear correlation was detected between CPSE and elastography values, these preliminary results evidence that the healthy prostates were characterized by a similar elastographic pattern, thus pointing that these techniques can be potentially useful to be applied in case of prostatic disorders to improve the accuracy of the final diagnosis in a non-invasive way.

Coat, Claw and Dewclaw 17-ß-Estradiol and Testosterone Concentrations in Male and Female Postpubertal Cats: Preliminary Results.

In the recent past, tissue materials such as hair/coat and nails/claws have proved to be useful for the study of long-term hormonal changes in humans and animals and shown to be advantageous in terms of being collectable without invasiveness, with a benefit in terms of animal welfare. However, studies using these tissue materials in cats are scarce, especially on sexual hormone measurement. In this study, the concentrations of 17-ß-estradiol (E2) and testosterone (T) were assessed in 20 male and 18 female domestic postpubertal cats at the time of neutering/spaying during the breeding season. Hormones were measured in coat shaved from the forearm (ACOAT) and from the surgical area (SCOAT); claws were collected from the forearms (CLAWS) and the dewclaws (DCLAWS). Although all these tissue materials were shown to be useful for E2 and T long-term measurement, only T concentrations were higher (p < 0.001) in males from both ACOAT and SCOAT samples when compared to females and therefore useful for distinguishing between the two sexes. Within each sex, E2 and T concentrations can be assessed on coat, but also on the dewclaws, providing an alternative, practical, matrix for sexual steroid measurement in postpubertal cats during the breeding season.

Treatment of a Recurrent Pyometra by Surgical Uterine Drainage in a Main Coon Cat.

Pyometra is a uterine disease typical of the luteal phase of the estrus cycle. For selected patients, such as breeding subjects, ovariohysterectomy is not a valid resolutive option. Medical treatments involving cloprostenol and aglepristone have been developed for the cats, but they can be ineffective in rare cases. Transcervical drainage and flushing have been described for the dogs, as well as for large wild cats. However, to the author's knowledge, there are no report of uterine drainage in cats. The present case describes an alternative treatment of pyometra in a 3-year-old Main Coon previously treated with aglepristone. The patient underwent a laparotomy: the uterus was exposed, and a sterile urinary catheter was inserted into each horn, through the wall of the uterus, to allow the drainage of pathological collection and a subsequent lavage with lukewarm sterile saline. Medical treatment with aglepristone and marbofloxacin was associated. After treatment, no recurrence was reported, and the cat had an uneventful pregnancy. Although it is a unique case report, the results presented are promising, as the technique appears to have provided healing and preserved fertility. Further studies are needed to confirm its efficacy in the long-term prevention of recurrence.

Management of Vaginal Hyperplasia in Bitches by Bühner Suture.

Vaginal hyperplasia in bitch is an exaggerated response of the vaginal mucosa to estrogens during the proestral-estral phase of the cycle that can protrude through the vulvar lips. The present study refers to the management of vaginal hyperplasia in bitches by Bühner vulvar suture. Fourteen private-owners animals were refereed for spontaneous vaginal hyperplasia and complete protrusion of the mucosa, without ischemic or necrotic areas, which occurred during the proestral-estral phase. Under general anesthesia, prolapsed mass was cleaned with 50% glucose solution to reduce oedema and gently repositioned; the Bühner suture was applied using a Gerlach needle and a sterile vaginal suture tape, maintaining a minimal opening to allow urination in order to avoid possible recurrence in the same estrus. None of the bitches showed recurrence during the current cycle, proving the effectiveness of the Bühner suture. To prevent the possible recurrence of vaginal hyperplasia at the subsequent estrus, all the bitches underwent an ovariectomy 2 months later, when the Bühner suture was removed. In conclusion, the Bühner suture proved to be useful for the conservative treatment of vaginal hyperplasia in medium- or large-sized bitches. However, this approach should be considered only for cases in which the prolapsed mass does not show trauma, ulceration, ischemic or necrotic areas.

Effectiveness of a Modified Administration Protocol for the Medical Treatment of Feline Pyometra.

Pyometra is a common uterine disease of dogs and cats, typical of the luteal phase. Traditionally, ovariohysterectomy was considered the elective treatment for pyometra, but in some cases, such as breeding subjects or patients with a high anesthesiologic risk, medical treatment is preferred. Aglepristone is a progesterone receptor blocker and its use proved to be effective for the medical treatment of pyometra in bitches and queens. The aim of this work is to report the effectiveness, in the feline species, of a modified aglepristone administration protocol. Five intact queens were referred to veterinary care centers for pyometra. Aglepristone (15 mg/kg) was administered at D0, D2, D5, and D8, as described by Contri and collaborators for dogs. An antibiotic treatment (marbofloxacin, 3 mg/kg) was associated, and uterine conditions were checked with regular ultrasonographic exams. The uterus returned to its normal condition 10 days after starting the treatment and no adverse effects were reported. After treatment, three queens had an uneventful pregnancy. Even if the treated group was restricted and homogeneous, the proposed modified protocol proved to be useful and promising for the medical treatment of pyometra in cats; further studies are planned to verify its effectiveness in the long-term prevention of recurrence.

Sentinel Lymph Node Mapping with Indirect Lymphangiography for Canine Mast Cell Tumour.

Mast cell tumour (MCT) is a common cutaneous and subcutaneous neoplasia in dogs. It can metastasise to lymph nodes (LNs), and this adversely affects the prognosis and treatment. The study aims to evaluate the SLN mapping of MCTs with radiographic indirect lymphography. Dogs that underwent clinical staging were prospectively enrolled. Lipiodol was injected around the MCT or the surgical scar. After 24 h, LNs that picked up contrast were radiographically assessed. Twenty-six dogs with 29 MCTs were included. MCTs were confirmed histologically, while SLNs were evaluated either by cytology and/or histology. SLNs were detectable in 23 dogs with 26 MCTs. Lymphatic vessels were visible in 19 MCTs. In nine MCTs, at least two SLNs picked up contrast. In particular, seven MCTs involved two SLNs, and two MCTs involved three different SLNs. In nine MCTs, at least a SLN was metastatic. This study indicates that the lymph drainage pattern of the MCTs may be different for each MCT, and more than one SLN can be involved. Indirect lymphangiography with Lipiodol allowed the detection of the SLN in 90% of MCTs. This provided clinically relevant information to remove the LN and stage the patient.

Ultrasound Examination of Unilateral Seminoma in a Salernitano Stallion.

An 18-year-old Salernitano stallion developed a progressive enlargement of the left testicle over eight months. An ultrasound evaluation was performed, along with a hormonal profile. A histopathological evaluation of the testis was performed after unilateral orchiectomy. On B-mode ultrasound examination, testicular parenchyma was characterized by the loss of internal structure, with the presence of multiple coalescing, nodular, well-defined and heterogeneous lesions with capsule deformity, appearing with an irregular profile. On dissection, the testicular parenchyma bulged over the cut section, confirming the increase in size. Microscopically, the lesion consisted mainly of large, densely packed, polygonal-to-round-shaped neoplastic cells. Mitotic figures were plentiful and frequently atypical; further microscopic features included apoptosis and necrosis. At immunohistochemistry, the entire neoplasm showed strong and diffuse immunolabeling for vimentin, while CD117-specific immunoreactivity was only observed in scattered clusters of neoplastic cells. Based on the gross, microscopic and IHC findings, a diagnosis of diffuse seminoma was made. Three months later, a follow-up examination showed no evidence of recurrence and the preservation of reproductive abilities. The case presented shows an unusual ultrasonographic pattern for seminoma and the basis of the correlation between the characteristics of the sonoelastographic examination and histological diagnosis.

The Lipidomics of Spermatozoa and Red Blood Cells Membrane Profile of Martina Franca Donkey: Preliminary Evaluation.

Fatty acid-based lipidomic analysis has been widely used to evaluate health status in human medicine as well as in the veterinary field. In equine species, there has been a developing interest in fertility and sperm quality. Fatty acids, being the principal components of the membranes, play an active role in the regulation of the metabolic activities, and their role on spermiogenesis seems to be of great importance for the resulting quality of the sperm and, thus, fertility. With the application of widely used lipidomic techniques, the aim of this study was to evaluate: (a) the fatty acid content of the spermatozoa's membranes of 26 healthy male Martina Franca donkeys and its possible correlation with sperm parameters, and (b) the evaluation of the composition of the red blood cells' membrane. PUFA omega-6 are the principal components (40.38%) of the total PUFA content (47.79%) in both types of cells; however, DPA is the predominant one on the spermatozoa's membrane (27.57%) but is not present in the erythrocyte's membrane. Spermatozoa's motility (%) is positively correlated with stearic acid and EPA, and progressive motility (%), with oleic acid. These findings offer information on the composition of both types of cells' membranes in healthy male MF donkeys and reflect the metabolic transformations of the spermatozoa's membrane during the maturation period, providing a better perception of the role of fatty acids in sperm parameters and fertility.

Maternal Phylogenetic Relationships and Genetic Variation among Rare, Phenotypically Similar Donkey Breeds.

The mitochondrial DNA (mtDNA) D-loop of endangered and critically endangered breeds has been studied to identify maternal lineages, characterize genetic inheritance, reconstruct phylogenetic relations among breeds, and develop biodiversity conservation and breeding programs. The aim of the study was to determine the variability remaining and the phylogenetic relationship of Martina Franca (MF, with total population of 160 females and 36 males), Ragusano (RG, 344 females and 30 males), Pantesco (PT, 47 females and 15 males), and Catalonian (CT) donkeys by collecting genetic data from maternal lineages. Genetic material was collected from saliva, and a 350 bp fragment of D-loop mtDNA was amplified and sequenced. Sequences were aligned and evaluated using standard bioinformatics software. A total of 56 haplotypes including 33 polymorphic sites were found in 77 samples (27 MF, 22 RG, 8 PT, 19 CT, 1 crossbred). The breed nucleotide diversity value (π) for all the breeds was 0.128 (MF: 0.162, RG: 0.132, PT: 0.025, CT: 0.038). Principal components analysis grouped most of the haplogroups into two different clusters, I (including all haplotypes from PT and CT, together with haplotypes from MF and RG) and II (including haplotypes from MF and RG only). In conclusion, we found that the primeval haplotypes, haplogroup variability, and a large number of maternal lineages were preserved in MF and RG; thus, these breeds play putative pivotal roles in the phyletic relationships of donkey breeds. Maternal inheritance is indispensable genetic information required to evaluate inheritance, variability, and breeding programs.

Cryotolerance of equine spermatozoa correlates with specific fatty acid pattern: A pilot study.

Sperm cryopreservation represents a powerful tool for horse breeding. To improve the efficiency of artificial insemination in the horse using cryopreserved spermatozoa, an adequate understanding of the underlying biophysical properties that affect sperm cryosurvival needs to be reached yet. In this pilot study, we described isolation and analysis of the main fatty acids from sperms of stallions classified as good and poor freezers (7 GF and 5 PF, according to sperm motility and viability, before and after cryopreservation). Fatty acid profiles were only assessed in pre-thaw sperms. Eight main fatty acids were identified, using gas chromatography, and their contents were expressed as percentage of the total lipid content. We found that lauric, myristic and oleic acid (C12:0, C14:0 and C18:1n9c) turned out to be about 2-fold more abundant in the sperm cells of the GFs compared with PFs. Moreover, we described for the first time the presence of a very high amount of a trans geometrical isomer of linoleic acid, linolelaidic acid (C18:2n6t), in pre-thaw PF spermatozoa. Notably, we found in fresh sperms of PF stallions a ratio of unsaturated fatty acids to saturated fatty acids which was twice that of those of GF group, suggesting a positive effect of a high saturated-to-unsaturated fatty acid ratio for the "freezability" of equine spermatozoa. Finally, principal component analysis (PCA) confirmed the relationships between specific fatty acids and cryotolerance of equine spermatozoa, also providing a graphical classification and additional information about the dominant variables governing the classification process.

Monolateral chronicon orchitis in a stallion.

Orchitis and epididymo-orchitis are inflammatory lesions of the testicle. We herein describe a case of monolateral chronic orchitis which occurred in a Tiro Pesante Rapido (TPR) stallion, born in 2002, with a history of good fertility. The stallion was healthy and asymptomatic although the left testis was found to be smaller as compared with the right one and was hard in consistency. Histopathology examination revealed tubular atrophy and parenchymal sclerosis. Scattered foci of calcification and chronic inflammation, the latter dominated by macrophages and lymphocytes, were also observed. Although lesions were clearly present, the semen was demonstraed to be of good quality. This study highlights the need for periodic clinical and ultrasound evaluation of stallions, in order to preserve their reproductive performance.