Preputial lymphangioma in a stallion: First report.

This short communication describes a case of a 30-year-old stallion with a mass on the base of the penis causing paraphimosis. The patient was submitted to anti-inflammatory and diuretic therapy with no signs of improvement, so 16 days after the lesion was detected, the animal was euthanized. Necropsy was performed, and histopathological assessment of the lesion was conducted. The mass was composed primarily of channels and cavernous structures, lined by elongated cells of vascular origin, located in the preputium. The lesion was diagnosed as a preputial lymphangioma. To the authors' best knowledge, the anatomical location of this neoplasm (which is rare in veterinary medicine) has not been previously reported.

Single Nucleotide Polymorphism in Prolactin Gene Is Associated With Clinical Aggressiveness and Outcome of Canine Mammary Malignant Tumors.

Prolactin (PRL) is a key hormone involved in canine mammary development and tumorigenesis. In this study, the influence of a single nucleotide polymorphism (SNP) in the PRL gene (rs23932236) on the clinicopathological parameters and survival of dogs with canine mammary tumors (CMTs) was investigated. A total of 206 female dogs with spontaneous mammary tumors were enrolled in this study and circulating blood cells were genotyped. This specific SNP was associated with larger size (>3 cm diameter) for malignant tumors (P = .036), tumors with infiltrative/invasive growth pattern (P = .010), vascular invasion (P = .006), and lymph node metastasis (P = .004). Carriers of the variant allele had a shorter overall survival compared to the wild-type population with an overall survival of 18.7 months and 22.7 months, respectively (P = .004). These findings suggest that SNP rs23932236 of canine PRL gene may be used as an indicator for the development of clinically aggressive forms of CMTs.

Estrogen receptors genotypes and canine mammary neoplasia.

BACKGROUND: Estrogens are essential for the development and proper function of several hormone-dependent organs. There are, however, several lines of evidence associating estrogens with mammary carcinogenesis. A marked individual genetic variability concerning estrogens biosynthesis, metabolism and mechanism of action was recognized and associated with human breast cancer susceptibility, clinical features and progression. Although some genetic variations in canine ESR1 gene were reported, their influence in clinicopathological features and progression of canine mammary tumors has not been fully evaluated. This study aims to assess the influence of SNPs in ESR1 gene (rs397512133, rs397510462, rs851327560, rs397510612, rs852887655, rs852684753 and rs852398698) in canine mammary tumors characteristics and progression. A group of 155 non-neutered bitches with mammary tumors was included in the study. Follow-up information was assessed 24 months after surgery. RESULTS: Genetic profiles associated with a later onset of mammary tumors and less aggressive clinicopathological features, namely smaller tumor size (≤ 3 cm) with extensive tubular differentiation and low canine-adapted prognostic index (vet-NPI), were identified in this study. CONCLUSIONS: Our data suggest that the ESR1 genetic profile may help on the decision regarding the selection of individual tailored preventive measures against canine mammary tumors development, such as early neutering.

Distribution of Inflammatory Infiltrate in Feline Mammary Lesions: Relationship With Clinicopathological Features.

Inflammation is a frequent finding in feline mammary neoplasms. Recent research suggests that the presence and location of tumour-associated immune cells might play a significant role in the clinical outcome of feline mammary carcinomas. The present study aimed to characterise the overall inflammatory infiltrates in healthy, hyperplastic/dysplastic, benign and malignant lesions of the feline mammary gland, and to evaluate its association with clinicopathological features. Perilesional and intralesional inflammatory foci were evaluated in 307 lesions from 185 queens, and categorised according to its distribution and intensity. The presence, location and density of tertiary lymphoid structures were also assessed. A control group included 24 queens without mammary changes. The presence of intralesional and perilesional inflammatory infiltrate was observed in a majority of the lesions (80.8% and 90.2%, respectively), but differed according to the type of mammary lesion, being more remarkable in malignant neoplasms. Only scarce individual cells were observed in 28.1% of the normal mammary glands. Data analysis revealed statistically significant associations (p < 0.05) between the presence of a more prominent intralesional and perilesional inflammatory reaction and several clinicopathological features associated with worse prognosis, including clinical stage, tumour size, mitotic count, lymphovascular invasion and lymph node metastasis. Furthermore, tertiary lymphoid structures were significantly more frequent in tumours with an infiltrative growth and lymph node metastasis. According to our results, the inflammatory reaction present in different types of feline mammary lesions is associated with the development of more aggressive tumours.

Mechanical stimulation-induced purinome priming fosters osteogenic differentiation and osteointegration of mesenchymal stem cells from the bone marrow of post-menopausal women.

BACKGROUND: Mechanical stimulation (MS) significantly increases the release of adenine and uracil nucleotides from bone marrow-derived mesenchymal stem cells (BM-MSCs) undergoing osteogenic differentiation. Released nucleotides acting via ionotropic P2X7 and metabotropic P2Y6 purinoceptors sensitive to ATP and UDP, respectively, control the osteogenic commitment of BM-MSCs and, thus, bone growth and remodelling. Yet, this mechanism is impaired in post-menopausal (Pm)-derived BM-MSCs, mostly because NTPDase3 overexpression decreases the extracellular accumulation of nucleotides below the levels required to activate plasma membrane-bound P2 purinoceptors. This prompted us to investigate whether in vitro MS of BM-MSCs from Pm women could rehabilitate their osteogenic commitment and whether xenotransplantation of MS purinome-primed Pm cells promote repair of critical bone defects in an in vivo animal model. METHODS: BM-MSCs were harvested from the neck of femora of Pm women (70 ± 3 years old) undergoing total hip replacement. The cells grew, for 35 days, in an osteogenic-inducing medium either submitted (SS) or not (CTR) to MS (90 r.p.m. for 30 min) twice a week. Increases in alkaline phosphatase activity and in the amount of osteogenic transcription factors, osterix and osteopontin, denoted osteogenic cells differentiation, while bone nodules formation was ascertain by the alizarin red-staining assay. The luciferin-luciferase bioluminescence assay was used to quantify extracellular ATP. The kinetics of the extracellular ATP (100 µM) and UDP (100 µM) catabolism was assessed by HPLC. The density of P2Y6 and P2X7 purinoceptors in the cells was assessed by immunofluorescence confocal microscopy. MS-stimulated BM-MSCs from Pm women were xenotransplanted into critical bone defects drilled in the great trochanter of femora of one-year female Wistar rats; bone repair was assessed by histological analysis 10 days after xenotransplantation. RESULTS: MS-stimulated Pm BM-MSCs in culture (i) release 1.6-fold higher ATP amounts, (ii) overexpress P2X7 and P2Y6 purinoceptors, (iii) exhibit higher alkaline phosphatase activity and overexpress the osteogenic transcription factors, osterix and osteopontin, and (iv) form larger bone nodules, than CTR cells. Selective blockage of P2X7 and P2Y6 purinoceptors with A438079 (3 µM) and MRS 2578 (0.1 µM), respectively, prevented the osteogenic commitment of cultured Pm BM-MSCs. Xenotransplanted MS purinome-primed Pm BM-MSCs accelerated the repair of critical bone defects in the in vivo rat model. CONCLUSIONS: Data suggest that in vitro MS restores the purinergic cell-to-cell communication fostering the osteogenic differentiation and osteointegration of BM-MSCs from Pm women, a strategy that may be used in bone regeneration and repair tactics.

Protective Effect of HER2 Gene Polymorphism rs24537331 in the Outcome of Canine Mammary Tumors.

The role of HER2 in canine mammary tumors is not completely elucidated, and the contradictory results published so far may be, in part, explained by the genetic variability recognized in the canine HER2 gene. Single nucleotide polymorphism (SNPs) in HER2 were recently associated with less aggressive canine mammary tumor histotypes. This study assesses the relationship between SNPs rs24537329 and rs24537331 in canine HER2 gene and clinicopathological characteristics and outcome of mammary tumors in a group of 206 female dogs. Allelic variants were observed in 69.8% and 52.7% of the dogs for SNP rs24537329 and rs24537331, respectively. Our results demonstrated that SNP rs24537331 was associated with decreased tumoral necrosis (HR: 3.09; p = 0.012) and with longer disease-specific overall survival (HR: 2.59; p = 0.013). However, no statistically significant associations were found between SNP rs24537329 and the tumors' clinicopathological characteristics or survival. Our data suggest that SNP rs24537331 may have a protective effect in canine mammary tumors, allowing the identification of a subgroup of animals prone to develop less aggressive forms of the disease. This study emphasizes the importance of the genetic tests associated with clinical images and histological examinations when assessing CMT outcomes.

An in-depth characterisation of European seabass intestinal segments for assessing the impact of an algae-based functional diet on intestinal health.

Sustainable farming of fish species depends on emerging new feed ingredients, which can alter the features of the digestive tract and influence animals' overall health. Recent research has shown that functional feeds hold great potential for enhancing fish robustness by evoking appropriate responses at the intestine level. However, there is a lack of extensive and accurate descriptions of the morphology of the gastrointestinal tract of most farmed fish. We have characterised the intestine of European seabass thoroughly, by targeting four segments - anterior, mid, posterior and rectum. Results indicated that the anterior segment is mostly associated with absorption-related features; this segment has the largest absorptive area, the longest villi, and the highest number of neutral goblet cells (GC). The posterior segment and rectum have distinct histomorphometric features, but both seem to be important for immunity, displaying the highest count of acid GC and the highest expression of immune-related genes. The strongest proliferating cell nuclear antigen (PCNA) signal was observed in the anterior intestine and rectum, with PCNA+ cells appearing at the base of the villi and the corresponding villi branches. We have also evaluated the impact of a novel feed supplemented with a macro- and microalgae blend and found that there were no differences in terms of growth. However, the alterations observed in the mid intestine of fish fed the blend, such as thickening of the submucosa and lamina propria, an increased number of leucocytes, and higher expression of immune- and oxidative stress-related genes, suggest that algae may have an immunomodulatory effect. In the current article, we have described the morphology and expression patterns of the intestine segments of European seabass in detail and have presented a comprehensive report of the indices and methods used for the semi-quantitative and quantitative histomorphometric assessments, thereby providing useful information for future studies that aim to maintain intestinal health through dietary interventions.

Retrospective study of canine cutaneous tumors submitted to a diagnostic pathology laboratory in Northern Portugal (2014-2020).

BACKGROUND: Cutaneous neoplastic diseases are the most and second-most frequently reported tumors in male and female dogs, respectively. The aims of this study were to report the occurrence of canine cutaneous tumors in a pathology laboratory located in Northern Portugal between 2014 and 2020, and to characterize and categorize the anatomical locations, breed, age, and sex of the animals affected with different types of neoplasms. RESULTS: Throughout the 7-year study, 1,185 cases were diagnosed as cutaneous tumors, with 62.9% being classified as benign, and 37.1% as malignant. Mast cell tumors (22.7%) were the most frequently diagnosed tumor type, followed by benign soft tissue tumors (9.7%), sebaceous gland tumors (8.1%), vascular tumors (7.9%) and soft tissue sarcomas (7.6%). Cutaneous tumors commonly exhibited multicentric occurrence (14.6%) followed by single occurrence in hindlimb (12.1%), forelimb (8.6%), buttock (7.1%), abdominal (6.5%) and costal (5.2%) areas. The odds of developing cutaneous neoplasia were higher with increasing age (p < 0.001). Females had an increased odds of developing skin tumors compared to males (crude OR = 2.99, 95% (2.51, 3.55); adj OR = 2.93, 95% (2.46, 3.49). Purebred dogs, as a group, showed a reduced odds of developing cutaneous tumors when compared to mixed-breed dogs (crude OR = 0.63, 95% (0.53, 0.74); adj OR = 0.75, 95% (0.62, 0.89). CONCLUSIONS: Mast cell tumors, benign soft tissue tumors and sebaceous tumors were the most common histotypes encountered. The epidemiological survey achieved with this study demonstrates the relative frequency of different types of tumors in this particular population. Furthermore, the results herein achieved can act as a basis or a beneficial reference for local veterinarians helping in the establishment of a preliminary and presumptive diagnosis of canine cutaneous tumors histotypes. Skin tumors are the most and second-most frequently reported tumors in male and female dogs, respectively. The aim of this study was to report the occurrence of canine skin tumors in a diagnostic pathology laboratory located in Northern Portugal, between 2014-2020 and to characterize the anatomical distributions, breed, age, and sex of the animals affected by different skin tumors. During this period, 1,185 cases were diagnosed as skin tumors; 62.9% were diagnosed as benign, while 37.1% were malignant. Mast cell tumors (22.7%) were the most frequently diagnosed neoplasia, followed by benign soft tissue tumors (9.7%), sebaceous gland tumors (8.1%), vascular tumors (7.9%) and soft tissue sarcomas (7.6%). Skin tumors commonly developed in more than one location (14.6%) followed by solitary development in hindlimb (12.1%), forelimb (8.6%), buttock (7.1%), abdominal (6.5%) and costal (5.2%) areas. An increased odds of developing skin neoplasms as the patient's age increase was detected. Females showed an increased odds in comparison to male dogs. Purebred dogs presented decreased odds for developing skin tumors in comparison to mixed-breed dogs. The information relevance achieved with this study demonstrates the relative frequency of different types of tumors in this particular population, acting as a basis or a beneficial reference for regional veterinarians when providing an initial diagnosis of canine skin tumors.

Fatal congenital and traumatic cervical spine injuries in a captive newborn plains zebra (Equus quagga).

Background: In this report, we describe the clinical, macro-, and microscopic findings of a newborn zebra victim of a fatal trauma and its possible causes are discussed in detail. Case Description: A plains zebra (Equus quagga) born in a Portuguese zoo was found in hypothermia and unable to get up. The animal was admitted to the hospital 36 hours later and, after the first hour of treatment and no attempts to get up, it began to convulse and died. At necropsy, subdural hematoma and atlantoaxial dislocation were the main findings. Conclusion: The inability to pinpoint the exact moment of the lesions' onset determines the importance of a thorough surveillance of the periparturient period and parturition of captive animals. Furthermore, literature concerning zebras, their reproduction, and neonatal period is scarce, reinforcing the need to report these cases.

Needle rinse cell blocks as an ancillary technique: Diagnostic and clinical utility in gastrointestinal neoplasia.

BACKGROUND: Fine-needle aspirate (FNA) cytology is often the first-choice method for diagnosing gastrointestinal nodular lesions. The FNA material can be converted to histopathology specimens by a needle rinse cell block (NRCB) technique, allowing ancillary studies to refine the cytologic diagnosis. Despite use in human pathology, NRCB has never been applied to canine or feline gastrointestinal neoplasia. OBJECTIVE: This study described NRCB methodology and its diagnostic utility in specific cases of neoplastic gastrointestinal lesions. METHODS: Needle rinses with saline were performed after ultrasound-guided FNAs of two intestinal lymphomas (canine and feline) and a canine gastrointestinal stromal tumor (GIST). The NRCB was prepared using the cell tube block technique and processed for paraffin embedding. Routine immunohistochemistry protocols (using CD3, PAX-5, and Ki-67 for lymphoma cases and vimentin, desmin, S-100, and KIT markers for GIST) were applied to NRCB sections, and the results were compared with matched tissue biopsies. RESULTS: NRCBs with adequate cell numbers, preservation, and good separation of blood were obtained. The diagnosis and immunophenotyping were confirmed in both cases of lymphoma in NRCBs. In the GIST, the immunolabeling of the neoplastic cells in NRCB was completely concordant with the tissue biopsy. CONCLUSIONS: The described methodology is suitable for veterinary settings, having few technical requirements and low invasiveness. The presented cases of gastrointestinal neoplasia highlight the utility of NRCBs as a platform to conduct ancillary studies and refine the cytologic diagnosis.

Vet-OncoNet: Malignancy Analysis of Neoplasms in Dogs and Cats.

Analysis of canine and feline tumor malignancy data can help clinicians identify high-risk patients and make more accurate decisions. Based on a sample of 16,272 cancer records, including 3266 cats and 13,006 dogs, collected from January 2019 to December 2021 in the Vet-OncoNet Network database, this study aimed to compare the tumor malignancy profile between cats and dogs, considering animal-related factors (sex, age, and breed), topography, and geographic location using a mixed-effects logistic regression model. Cats had a higher proportion of malignant tumors (78.7%) than dogs (46.2%), and the malignancy profile was very different regarding tumors' topographies. The mean age of malignant tumors occurred eight months later than benign ones (9.1, SD = 3.4; 9.8, SD = 3.2), in general. Species (OR = 3.96, 95%CI 3.57: 4.39) and topography (MOR = 4.10) were the two most important determinants of malignancy risk. Female dogs had a higher risk than male dogs (OR = 1.19, 95%CI 1.08: 1.31), which does not appear to be the case in cats (OR = 0.98, 95%CI 0.77: 1.23). Breed contributed significantly to differences in malignancy risk in dogs (MOR = 1.56), particularly in pit bulls and boxers. District of residence was not so relevant in predicting malignancy risk (MOR = 1.14). In both species, the risk of malignancy increased by approximately 20% every three years. It could be hypothesized that species differences in genetic structure may contribute to tumor malignancy.

Cross Species Analysis and Comparison of Tumors in Dogs and Cats, by Age, Sex, Topography and Main Morphologies. Data from Vet-OncoNet.

The animal cancer burden is essential for the translational value of companion animals in comparative oncology. The present work aims to describe, analyze, and compare frequencies and associations of tumors in dogs and cats based on the Animal Cancer Registry created by Vet-OncoNet. With 9079 registries, regarding 2019 and 2020, 81% (n = 7355) belonged to dogs. In comparison, cats have a general one-year right advance in the mean age of cancer diagnosis compared to dogs. The multivariate topography group analysis shows a distinct pattern between the two species: dogs have higher odds of cancer in the genito-urinary system, spleen, soft tissue tumors and skin, while cats show higher odds for tumors in the eyes, digestive organs, nasal cavity, lymph nodes, bones and mammary glands. Regarding morphologies, dogs are overrepresented in mast cell tumors (MCT), melanomas, and hemangiosarcomas. While cats are overrepresented in fibrosarcomas, lymphomas (T and B-cell), in malignant mammary tumors, and squamous cell carcinoma (SCC). Females have greater odds only in the mammary gland, with males having greater odds in six of twelve topographies. This study is the first outcome of continuous animal cancer registration studies in Portugal.

Adjuvant doxorubicin vs metronomic cyclophosphamide and meloxicam vs surgery alone for cats with mammary carcinomas: A retrospective study of 137 cases.

This study aims to evaluate the efficacy and side effects of low dose cyclophosphamide chemotherapy plus meloxicam as an adjuvant treatment, compared with high dose doxorubicin or surgery alone in cats with mammary carcinoma. Medical records of 228 female cats treated for mammary carcinoma between 2008 and 2018, were reviewed in eight veterinary institutions. Only cats with complete tumour staging and radical mastectomy were included in the study. One hundred and thirty-seven cats were divided into three treatment groups: group 1 (n = 80) cats treated with surgery, group 2 (n = 34) cats that had surgery and adjuvant treatment with doxorubicin, and group 3 (n = 23) cats with surgery and adjuvant treatment with low dose metronomic cyclophosphamide and meloxicam. The study endpoints were disease free interval (DFI) and overall survival (OS). Toxicity was evaluated according to the VCOG-CTCAE criteria. The median DFI was 270, 226 and 372 days in groups 1, 2 and 3, respectively. The median OS was 338 (group 1), 421 (group 2) and 430 (group 3) days. The differences between groups were not significant (DFI P = .280 and OS P = .186). Toxicity was observed in 52.9% (n = 18) of cats in group 2 and 39.1% (n = 9) of cats in group 3, with mild to moderate intensity. Differences were not significant (P = .306). In conclusion, adjuvant chemotherapy treatment did not improve survival and the overall benefit remains unproven. Randomized prospective trials are necessary to clarify the effectiveness of adjuvant chemotherapy treatment for feline mammary carcinomas.