Bleaching melanin in formalin-fixed and paraffin-embedded melanoma specimens using visible light: a pilot study

In fluorescence microscopy, light radiation can be used to bleach fluorescent molecules in formalin-fixed and paraffin-embedded (FFPE) samples, in order to increase the ratio between signal of interest and background autofluorescence. We tested if the same principle can be exploited in bright field microscopy to bleach pigmented melanoma FFPE sections together with cell morphology maintenance. After dewaxing and rehydration, serial FFPE sections of a feline diffuse iris melanoma, a canine dermal melanoma, a gray horse dermal melanoma and a swine cutaneous melanoma were irradiated with visible light for I, 2, 3, 4 and 5 days, prior to Hematoxylin & Eosin staining. Complete bleaching was obtained after 1-day treatment in feline and swine melanomas, while 2 and 3 days were required in canine and equine neoplasms, respectively. In all treated samples, cell morphology was maintained. Photo-induced bleaching combined with immunohistochemistry was tested after a 3-day photo-treatment using five different markers. According to the literature, in all samples neoplastic cells stained positive for vimentin, S100 and PNL2, while negative for FVIII and pancytokeratin. in conclusion, visible light can be effectively exploited to bleach pigmented melanoma FFPE sections prior to perform routine histochemical and immunohistochemical stains

Active surveillance of paratuberculosis in Alpine-dwelling red deer (Cervus elaphus)

Paratuberculosis (Johne’s disease) is a globally widespread infectious disease affecting domestic and wild ruminants, caused by Mycobacterium avium subsp. paratuberculosis (MAP). The bacterium is excreted in the feces and is characterized by high environmental resistance. The new Animal Health Law (Regulation EU 2016/429) on transmissible animal diseases, recently in force throughout the European Union, includes paratuberculosis within the diseases requiring surveillance in the EU, listing some domestic and wild Bovidae, Cervidae, and Camelidae as potential reservoirs. Taking advantage of a culling activity conducted in the Stelvio National Park (Italy), this study investigated MAP infection status of red deer (Cervus elaphus) between 2018 and 2022, and evaluated the probability of being MAP-positive with respect to individual and sampling-level variables. A total of 390 subjects were examined macroscopically and tested for MAP, using different diagnostic tools: IS900 qPCR, culture, histopathology, and serology. Twenty-three of them were found positive for MAP by at least one test, with an overall prevalence of 5.9% (95% CI 4.0–8.7), that, respectively, ranged from 12.4% in the first culling season to 2.0 and 2.1% in the 2019–2020 and 2021–2022 culling seasons. Quantitative PCR assay on ileocecal valve and mesenteric lymph nodes detected the highest number of MAP positive animals. The results of the study showed the increased probability of being MAP-positive with increasing age and that red deer with lower body mass values were more likely to be infected with MAP. Overall, the absence of signs of clinical paratuberculosis and gross lesions together with the low level of shedding witness early phases of the disease among the positive red deer and support an improvement of the paratuberculosis status of this population, as shown by the decreased prevalence of the disease over the years

Feline Visceral Mycobacteriosis

Mycobacteria in cats are responsible for cutaneous or visceral disease, often with systemic involvement (Gunn-Moore et al., 2011). Infection can be sustained by tuberculous (e.g. M.bovis) and non-tuberculous mycobacteria (e.g. M.avium and M.lepraemurium) (Lee et al., 2017). The aim of this study is to evaluate the distribution of the lesions in feline visceral mycobacteriosis. Twenty-nine necropsy cases of feline visceral mycobacteriosis, conferred from 1965 to 2017, were studied. On histopathology hematoxylin-eosin and acid-fast stains were performed. Mycobacteria strains were identified by microbiological and molecular methods. Twenty-three out of 29 cases were submitted for necropsy in autumn and winter. When breed was known: nine cats were Persian, 8 Siamese and 10 crossbreed. In 17 out of 24 cases where age was known, it was lesser than 5 years. Two cases had lesions confined to the digestive system, 3 to the respiratory system, while 24 cases were systemic forms. Thirteen cases out of 29 were identified: 2 as Mycobacterium spp, 7 as M.bovis and 4 as M.avium. Siamese, Persian breeds and young cats were overrepresented as already described (Gunn-Moore, 2014). The occurrence of mycobacterioses in our cases was higher during cold seasons, contrary to the man in which tuberculosis is a long term localized disease, more often diagnosed in summer even if acquired in winter; in cat the disease tends to generalize making shorter the course of the disease (Fares, 2011). This is confirmed by lesions’ distribution that indicates a greater occurrence of systemic compared to localized forms. Features of gastrointestinal lesions indicate the alimentary as the primary route of infection (Fig.1). Since the most recent cases were sustained exclusively by M.avium, while older cases were caused by M.bovis, a switching in the most diffused species or a different source of infection might occurred in the last years (Pesciaroli et al., 2014)

Chylopericardium Effusion in a Lac Alaotra Bamboo Lemur (<i>Hapalemur alaotrensis</i>)

An 11-year-old female Hapalemur alaotrensis was evaluated following a history of dyspnea of 15 days’ duration. Thoracic radiography performed by the referring veterinarian revealed a large cardiac silhouette and dorsal deviation of the trachea. Heart sounds were muffled. Echocardiographic findings were indicative of severe pericardial effusion without cardiac tamponade. No pleural effusion was identified. A computed tomography (CT) exam confirmed the presence of severe pericardial effusion and allowed identification of a parenchymatous mediastinal lesion sited at the level of the left hemithorax. To delineate the thoracic duct, lymphoCT was also performed by injection of iodinated contrast medium in the perianal subcutaneous tissue. Pericardiocentesis yielded a considerable amount of effusion with chylous biochemical and cytological properties. A diagnosis of chylopericardium with absence of pleural effusion was made. Initially, the chylopericardium was managed conservatively with two centesis and oral treatment with prednisolone. Medical treatment did not result in complete resolution of effusion and clinical signs; therefore, subtotal pericardiectomy and thoracic duct ligation were recommended. After the second pericardiocentesis, the subject died and the pericardiectomy could not be performed. To the authors’ knowledge, this is the first report of the development of chylopericardium in a Hapalemur alaotrensis

Ovarian Neuroglial Choristoma in a Bitch

Neuroglial choristomas are rare malformations consisting of heterotopic mature neural tissue at a site isolated from the brain or spinal cord. In human medicine, neuroglial choristomas are predominantly reported in the head and in the neck, except for one recent case reported in a foot of a child. In domestic animals, neuroglial choristomas are exceedingly rare, reported only in the retina of a dog, in the pharynx and in the skin of two kittens, and within the oropharynx of a harbor seal. A three-year-old intact female Jack Russell Terrier presented for elective ovariectomy exhibited a cystic lesion 2 cm in diameter expanding in the right ovary. Histological examination of the lesion revealed a mass composed of well-organized neuroglial tissue. Immunohistochemistry with primary antibodies against GFAP, NSE, and IBA-1 confirmed the neuroglial origin of the mass. At the time of this writing, 7 years after ovariectomy, the dog was clinically normal. Together with a recent case described in the foot of a child, this case confirms that neuroglial choristoma may also be found far from the skull or spine, supporting the hypothesis that they may arise from an early embryological migration defect

Peripheral Nerve Sheath Tumors Resembling Human Atypical Neurofibroma in Goldfish (Carassius auratus, Linnaeus, 1758)

Skin spindle cell tumors (SSTs) frequently occur in fishes, with peripheral nerve sheath tumors (PNSTs) being the most commonly reported neoplasms in goldfish. However, distinguishing PNSTs from other SCTs is not always possible when relying exclusively on routine cytological and histopathological findings. Therefore, the aim of this study is to characterize six skin nodules, resembling atypical neurofibromas in humans, found in six cohabiting goldfish (Carassius auratus), and to determine a minimal subset of special stains required to correctly identify PNSTs in this species. Routine cytology and histopathology were indicative of an SCT with nuclear atypia in all cases, with randomly distributed areas of hypercellularity and loss of neurofibroma architecture. Muscular and fibroblastic tumors were excluded using Azan trichrome staining. Alcian blue and Gomori’s reticulin stains revealed the presence of intratumoral areas of glycosaminoglycans or mucins and basement membrane fragments, respectively. PAS and PAS–diastase stains confirmed the latter finding and revealed intra- and extracellular glycogen granules. Immunohistochemistry displayed multifocal, randomly distributed aggregates of neoplastic cells positive for S100 protein and CNPase, intermingled with phosphorylated and non-phosphorylated neurofilament-positive axons. Collectively, these findings are consistent with a PNST resembling atypical neurofibroma in humans, an entity not previously reported in goldfish, and suggest that Azan trichrome staining, reticulin staining, and immunohistochemistry for S100 protein and CNPase represent a useful set of special stains to identify and characterize PNSTs in this species

Lymphatic invasion is a significant indicator of poor patient outcome in canine bladder urothelial carcinoma

Background: Urothelial carcinoma (UC), also known as transitional cell carcinoma, is the most common malignant tumor of the canine urinary bladder and represents a model for studying human bladder cancer. However, the existing literature has limited data on the clinicopathological characteristics of these tumors and their prognostic value. Aim: The aim of this study was to evaluate such factors, correlating them with follow-up, in a group of 32 dogs with bladder UC. Methods: Clinical data of these cases, submitted to São Paulo State University and VetPat Private Laboratory (São Paulo/Brazil), were recorded between January 2000 and November 2019. For each case, formalin-fixed and paraffinembedded sections were stained with hematoxylin and eosin and histologically evaluated. Survival analysis was carried out, and the prognostic value of the presence of lymphatic invasion and the treatment used was determined. Results: Dogs with neoplastic lymphatic vessel invasion had a lower overall survival compared to those without lymphatic invasion, and dogs that received vinblastine in addition to surgery had higher global survival when compared to animals that received carboplatin in addition to surgery.Conclusion: The results obtained show the importance of further studies regarding the prognostic value of the two factors demonstrated as potential survival predictors, especially the lymphatic vessel invasion

Data_Sheet_1_Active surveillance of paratuberculosis in Alpine-dwelling red deer (Cervus elaphus).PDF

Paratuberculosis (Johne’s disease) is a globally widespread infectious disease affecting domestic and wild ruminants, caused by Mycobacterium avium subsp. paratuberculosis (MAP). The bacterium is excreted in the feces and is characterized by high environmental resistance. The new Animal Health Law (Regulation EU 2016/429) on transmissible animal diseases, recently in force throughout the European Union, includes paratuberculosis within the diseases requiring surveillance in the EU, listing some domestic and wild Bovidae, Cervidae, and Camelidae as potential reservoirs. Taking advantage of a culling activity conducted in the Stelvio National Park (Italy), this study investigated MAP infection status of red deer (Cervus elaphus) between 2018 and 2022, and evaluated the probability of being MAP-positive with respect to individual and sampling-level variables. A total of 390 subjects were examined macroscopically and tested for MAP, using different diagnostic tools: IS900 qPCR, culture, histopathology, and serology. Twenty-three of them were found positive for MAP by at least one test, with an overall prevalence of 5.9% (95% CI 4.0–8.7), that, respectively, ranged from 12.4% in the first culling season to 2.0 and 2.1% in the 2019–2020 and 2021–2022 culling seasons. Quantitative PCR assay on ileocecal valve and mesenteric lymph nodes detected the highest number of MAP positive animals. The results of the study showed the increased probability of being MAP-positive with increasing age and that red deer with lower body mass values were more likely to be infected with MAP. Overall, the absence of signs of clinical paratuberculosis and gross lesions together with the low level of shedding witness early phases of the disease among the positive red deer and support an improvement of the paratuberculosis status of this population, as shown by the decreased prevalence of the disease over the years.</p