Davide De Biase

Histological classification of mammary proliferative lesions in dogs and cats is essential for prognostic and predictive studies, as well as for comparative oncology; however, standardization of diagnoses is highly affected by substantial interobserver variability. In this study, a structured diagnostic algorithm based on the Davis–Thompson Foundation classification was assessed for the ability to improve interobserver agreement in the morphological diagnoses of mammary gland lesions in dogs and cats. Consensus diagnoses of 114 selected mammary lesions (91 canine, 23 feline) made by 16 veterinary pathologists with mammary tumor expertise were used as the gold standard, and 79 veterinary pathologists from 21 countries evaluated digitized whole slides in 2 rounds: a first reading without the algorithm and a second reading with the algorithm. Interobserver concordance was measured using unweighted kappa statistics for lesion category (hyperplastic/dysplastic, benign, malignant) and histological subtype. Across all lesions, agreement was higher for lesion category than for histological subtype. A slight improvement in concordance was observed when applying the algorithm, both for category (k = 0.72, before, and k = 0.73, after) and histotype classifications (k = 0.40, before to k = 0.42, after). Also, by using the algorithm, 73/79 raters (92%) modified at least 1 diagnosis and, among these, approximately two-thirds reached agreement with the gold standard in at least 1 case. Considering histotypes, malignant tumors with mixed epithelial and myoepithelial components showed persistently low agreement. Overall, the algorithm provided a reproducible diagnostic framework that may support training and improve standardization of canine and feline mammary tumor classification.

Corneal wound healing largely depends on the activation of stromal fibroblasts, which coordinate extracellular matrix remodeling and tissue regeneration. Although advanced platelet-rich fibrin (A-PRF+) has emerged as a promising biomaterial in regenerative medicine, its biological effects on equine corneal fibroblasts remain poorly understood. In this study, primary equine corneal fibroblasts were isolated from stromal explants and treated with an A-PRF+-conditioned medium. Cell metabolic activity, viability, intracellular reactive oxygen species, wound healing, cell cycle progression, growth factor secretion, and collagen type I expression were evaluated using colorimetric assays, fluorescence imaging, flow cytometry, and ELISA. A-PRF+ significantly enhanced fibroblast metabolic activity, increased intracellular ROS production, accelerated wound closure and cell cycle re-entry, and stimulated the secretion of PDGF-BB, TGF-β1, and VEGF-A. Furthermore, treatment increased the percentage of collagen type I-positive fibroblasts, supporting enhanced extracellular matrix production. Collectively, these findings demonstrate that A-PRF+ exerts beneficial effects on primary equine corneal fibroblasts by stimulating their metabolic, proliferative, migratory, and secretory activities. These results provide an experimental basis for the use of A-PRF+-based eye drops as a regenerative strategy to improve corneal healing in horses.

Canine cutaneous neoplasms are common and morphologically heterogeneous lesions whose diagnosis relies on integrating gross examination, cytology, and histopathology. This retrospective pilot study assessed the feasibility of a multimodal GPT-based large language model as an assistive, not autonomous, tool for standardized description, differential diagnosis generation, and classification support across this diagnostic workflow. Fifty-one histologically confirmed canine cutaneous tumors were retrospectively selected from the laboratory information system of the Veterinary Pathology Laboratory, University of Naples Federico II. For each case, de-identified gross photographs, digitized cytology, and representative histologic images were provided to the model using templated prompts. Model outputs were independently reviewed by two veterinary pathologists, who reached consensus on descriptive quality and diagnostic concordance with the histologic reference diagnosis. Final diagnostic outputs were classified as correct, partially correct, or incorrect. Strict accuracy was defined as the proportion of fully correct diagnoses, whereas broad accuracy combined correct and partially correct outputs considered diagnostically informative. Overall, the model achieved a strict diagnostic accuracy of 66.7% (34/51; 95% CI: 53.0–78.0) and a broad diagnostic accuracy of 90.2% (46/51; 95% CI: 79.0–95.7). Performance was highest in epithelial tumors and lower in mesenchymal and melanocytic tumors, in which the model more often identified broader diagnostic categories than specific histotypes. These findings suggest that GPT-based systems may support report standardization, descriptive consistency, and morphology-driven reasoning in veterinary pathology. However, reduced entity-level specificity, variable descriptive quality, and the risk of plausible but non-concordant outputs require strict human supervision and further validation before routine implementation.

The term asphyxia refers to a disruption in brain function due to rapid and persistent cerebral hypoxia or anoxia as a consequence of accidental or non-accidental injury. Considering the different mechanisms that may determine asphyxiation, such injuries can be referred to different categories: strangulation (death by hanging, ligature or manual strangulation), suffocation (smothering, choking, confined spaces and vitiated atmosphere), mechanical asphyxia (positional and traumatic asphyxia) and drowning (submersion or immersion in liquid). In both human and veterinary forensic practice, fatal asphyxia is considered among the most diagnostically challenging categories of sudden death, as it often produces only subtle and non-pathognomonic macroscopic signs, which can be easily covered by post-mortem alterations. Therefore, a wide range of information is often needed for the diagnosis of asphyxiation, including medical history, crime scene analysis, testimonies and physical evidence, along with the macroscopic and histological findings. The following review addresses the main lesions, ancillary tests and diagnostic issues associated with non-drowning asphyxia in veterinary forensic pathology.

A comparative analysis of testicular tumors in dogs and humans highlights notable similarities in cancer behavior, particularly regarding environmental influences. The ubiquitous distribution of endocrine disruptors in industrialized countries and their effects are strongly linked to the development of testicular cancers. This study examined 221 and 174 cases of testicular tumor in dogs and humans, respectively, from the Campania Animal Cancer Registry and the database of the National Cancer Institute “Fondazione G. Pascale-IRCCS”, between 2020 and 2023. These data were integrated with environmental data from the Ministry of Environment and Energy Security website and the Campania Regional Agency for Environmental Protection (ARPAC). The areas with the highest frequency of testicular tumors in both species were the municipalities of Napoli, Salerno, and Caserta, and their corresponding provinces. The same locations experienced severe contamination of soil and water by environmental pollutants, probably correlated to testicular tumors. Although the molecular mechanisms are not yet fully understood, our data suggest a strong link between the frequency of testicular tumors in humans and dogs and high concentrations of environmental pollutants in the reported locations. These findings emphasize the value of dogs as a potential animal model for studying human health and the effect of environmental factors on cancer development.

Flexural deformities (FDs) are a common condition in foals. Therapy is typically initiated without a precise diagnosis, and the etiopathogenesis often remains unknown. This study aimed (1) to investigate the clinical and pathological findings in congenital FD cases in foals and (2) to retrospectively describe the abnormalities detected in muscle biopsies of foals affected by CFDs. For these purposes, a retrospective study of the findings of muscle biopsies taken from foals with FDs referred to the Department of Veterinary Medicine and Animal Production at the University of Naples Federico II was performed from January 2005 to February 2024. Anamnesis, physical examination, hematological and biochemical data, along with the findings of histopathological muscle biopsy analysis and follow-up, were recorded. The clinical records of 15 cases of FDs were evaluated. The main clinical symptoms included flexural limb deformities associated with weakness, torticollis and scoliosis, mandibular prognathism, and inferior eyelid entropion. The evaluation of histopathological reports allowed us to observe the following muscle disorders: Core-like myopathy, mild nonspecific myopathy, mitochondrial myopathy, congenital fiber type disproportion, lipid storage myopathy, lipomatous dystrophy, myopathy with inclusion bodies, polysaccharide storage myopathy, and neurogenic myopathy. Even though many cases of FDs were diagnosed through clinical examination and successfully treated, we hypothesize that different underlying etiologies may present with similar flexural symptoms. A better understanding of these underlying causes is, therefore, desirable. These findings suggest that histopathological analysis may be a valuable tool for investigating FDs in foals, although further studies are needed to evaluate the significance of the observed alterations.

Several reports have indicated that canine MCTs express a mutated form of a tyrosine kinase receptor, namely KIT, that is involved in abnormal mast cell growth and differentiation. Currently, the post-surgical prognosis for MCTs is related to three different KIT immunohistochemical expression patterns. However, to our knowledge, there are few studies specifically exploring the efficacy of treatment with tyrosine kinase inhibitors related to KIT staining pattern. The purpose of this study was to investigate the potential theragnostic role of KIT expression patterns by studying their correlation to the overall survival and progression-free survival in dogs treated with only tyrosine kinase inhibitors immediately after surgery. We selected 66 cases of canine cutaneous MCTs with complete clinical background. A statistical analysis was performed to assess the overall survival status. Our data suggest an important role of KIT in the etiopathogenesis of canine MCTs and indicate that the anomalous cytoplasmatic distribution of KIT is potentially related to a lower efficacy of tyrosine kinase inhibitors, thus providing a significant prognostic information about the treatment outcome.

Gunshot wound morphology and gunshot residues (GSRs) evaluation have been poorly investigated in veterinary forensic pathology. The aims of the present study were to assess the gunshot wound morphology in animals and evaluate the detectability of lead deriving from GSRs using colorimetric techniques. To these aims, cadavers were divided into four different groups. Group A comprised eight animals who died from firearm-related injuries, while groups B and C included dog limbs shot using different shooting ranges; group D comprised dog limbs stabbed with a screwdriver. Morphological analysis was performed on all entry gunshot wounds. Lead residues were investigated using a Bullet Hole Testing Kit (BTK) and Rhodizonate Sodium histochemical staining (NaR-s). Gunshot wounds in group A showed an abrasion ring associated with hemorrhages and tissue necrosis. Groups B and C showed injuries related to the shooting range. NaR-s showed positive results in both animals that died from gunshot wounds and experimentally shot limbs. However, the number of positive cases and the pattern of lead distribution varied with the shooting range. Positive results by BTK were limited to close-contact shots in group B limbs. Our results suggest that both pathological examination and NaR tests represent valid tools for investigating gunshot wounds in veterinary pathology.

Fire-related deaths are usually a consequence of carbon monoxide (CO) poisoning or shock from thermal injuries. In humans, high levels of carboxyhemoglobin (COHb) concentrations in the blood can support a diagnosis of CO poisoning. In veterinary medicine, few studies investigated the pathological changes and blood COHb% in fire victims, and no data are available on post-mortem changes in blood gas composition due to fire. This study aims to investigate the pathological changes and COHb levels in both animal victims of fire and cadavers experimentally exposed to fire. For this purpose, dogs were selected and subdivided into three groups. Group A comprised 9 adult dogs, and Group B comprised 7 puppies that died under fire-related conditions. Group C was represented by 4 dog cadavers experimentally exposed to heat and smoke. A complete macroscopic, histological, and COHb evaluation were performed on each animal. Animals in Groups A and B showed cherry-red discoloration, thermal-injuries and soot deposits along the respiratory tract. Animals in Group C showed thermal injuries and soot deposits limited to the upper respiratory tract. The mean COHb% values in cadavers in Group C were lower than those observed in the other groups but higher compared to the values detected before the heat and smoke treatment. These findings suggest that both pathological changes and COHb analysis are valid tools for investigating fire-related deaths in dogs. However, the increase of COHb levels in cadavers exposed post-mortem to heat and smoke highlights how the COHb analysis should always be evaluated together with macroscopical and microscopical findings to avoid significant misjudgments in investigating fire-related fatalities in veterinary forensic practice.

Coronaviruses are highly transmissible and pathogenic viruses for humans and animals. The vast quantity of information collected about SARS-CoV-2 during the pandemic helped to unveil details of the mechanisms behind the infection, which are still largely elusive. Recent research demonstrated that different class I/II human leukocyte antigen (HLA) alleles might define an individual susceptibility to SARS-CoV-2 spreading, contributing to the differences in the distribution of the infection through different populations; additional studies suggested that the homolog of the HLA in cats, the feline leukocyte antigen (FLA), plays a pivotal role in the transmission of viruses. With these premises, this study aimed to exploit a bioinformatic approach for the prediction of the transmissibility potential of two distinct feline coronaviruses (FCoVs) in domestic cats (feline enteric coronavirus (FeCV) and feline infectious peritonitis virus (FIPV)) using SARS-CoV-2 as the reference model. We performed an epitope mapping of nonapeptides deriving from SARS-CoV-2, FeCV, and FIPV glycoproteins and predicted their affinities for different alleles included in the three main loci in class I FLAs (E, H, and K). The predicted complexes with the most promising affinities were then subjected to molecular docking and molecular dynamics simulations to provide insights into the stability and binding energies in the cleft. Results showed the FLA proteins encoded by alleles in the FLA-I H (H*00501 and H*00401) and E (E*01001 and E*00701) loci are largely responsive to several epitopes deriving from replicase and spike proteins of the analyzed coronaviruses. The analysis of the most affine epitope sequences resulting from the prediction can stimulate the development of anti-FCoV immunomodulatory strategies based on peptide drugs.