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Simple Summary: Molecular profiling of cancer is used to examine for specific mutations or proteins and is becoming increasingly important in oncology. An example of such a mutation is the altered BRAF gene and the resulting changed BRAF protein, which is already known to play a role in urinary bladder and prostatic carcinomas in dogs. We assumed that the mutant BRAFV595E protein should be considered in other canine carcinomas and tested 227 samples of 11 different carcinoma origins (anal sac (n = 23), intestine (n = 21), liver (n = 21), lungs (n = 19), mammary gland (n = 20), nasal cavity (n = 21), oral epithelium (n = 18), ovary (n = 20), prostate (n = 21), thyroid gland (n = 21), urinary bladder (n = 22)). We used immunohistochemistry with two different primary antibodies, each binding to the altered BRAF protein, and confirmed our results with droplet digital PCR (ddPCR). Among all the tested canine carcinomas, we found BRAF-mutated tumours in the prostate (16/21), the urinary bladder (17/22), and the oral cavity epithelium (4/18), while other carcinomas tested negative. Our findings showed that both antibodies are dependable tools for detecting the BRAFV595E mutation in canine carcinomas. Detecting BRAF mutations is important for applying future therapeutic approaches, including BRAF inhibitors. Alterations of the BRAF gene and the resulting changes in the BRAF protein are one example of molecular cancer profiling in humans and dogs. We tested 227 samples of canine carcinomas from different anatomical sites (anal sac (n = 23), intestine (n = 21), liver (n = 21), lungs (n = 19), mammary gland (n = 20), nasal cavity (n = 21), oral epithelium (n = 18), ovary (n = 20), prostate (n = 21), thyroid gland (n = 21), urinary bladder (n = 22)) with two commercially available primary anti-BRAFV600E antibodies (VE1 Ventana, VE1 Abcam). The immunohistochemical results were confirmed with droplet digital PCR (ddPCR). BRAFV595E-mutated cases were found in canine prostatic (16/21), urothelial (17/22), and oral squamous cell carcinomas (4/18), while other carcinoma types tested negative. Both antibodies showed consistent results, with intracytoplasmic immunolabeling of tumour cells, making them reliable tools for detecting the BRAFV595E mutation in canine carcinomas. In conclusion, identifying BRAF mutations from biopsy material offers a valuable opportunity to enhance cancer treatment strategies (BRAF inhibitors) in canine urothelial carcinomas, prostatic carcinomas, and oral squamous cell carcinomas. [ABSTRACT FROM AUTHOR]

Simple Summary: A dog imported from Greece to Germany presented a mass in the mammary gland. Cytology of the secretion was performed, and besides the malignant epithelial cells, amastigotes of Leishmania (L.). infantum were identified. Diagnoses of mammary carcinoma and canine leishmaniasis were confirmed by histopathology, PCR testing, and serology. The detection of L. infantum amastigotes in the mammary glands indicates a possible shedding by the milk and makes trans-mammary transmission to puppies theoretically possible as another route of L. infantum transmission apart from vectorial transmission by sand flies, blood transfusion, vertical transmission, and venereal transmission. Mammary tumors are one of the most common neoplasms in female dogs, and cytology represents a non-invasive diagnostic method. The protozoal pathogen Leishmania spp. was previously demonstrated in canine mammary glands. An eight-year-old, female-spayed Doberman was imported from Crete, Greece, three years before the first presentation. The dog was presented due to a mammary tumor two years after adoption. The clinical examination revealed fever and weight loss. Smears of the mammary secretion were investigated cytologically. Multiple atypical epithelial cells with moderate to marked criteria of malignancy were detected. Furthermore, amastigotes were visible intra- and extracellularly. The diagnosis of L. infantum infection was based on a positive PCR out of the cytological smear, and a positive serology. Mammary carcinoma and granulomatous inflammation with amastigotes were confirmed by histopathology. We aimed to provide a complete report of cytological, histopathological, hematological, and biochemistry findings in a dog with L. infantum in the mammary glands with focus on trans-mammary pathogen transmission as a potential alternative way of spreading Leishmania infections. Canine leishmaniasis should be a potential differential diagnosis in dogs with lesions and/or inflammation in the mammary glands and a history of presence in areas endemic for L. infantum, especially the Mediterranean in Europe. [ABSTRACT FROM AUTHOR]

Simple Summary: Chalkbrood is a mycological brood disease of the Western honey bee (Apis mellifera), caused by the fungus Ascosphaera apis. To date, the course of this disease (pathogenesis) has not been investigated in depth. In this study, the pathogenesis will be described in detail using macroscopy and histology under controlled experimental conditions. For this purpose, Apis mellifera larvae were artificially reared and experimentally infected with Ascosphaera apis spores. The infected larvae were then evaluated macroscopically and histologically, and the findings were compared to non-infected control larvae. Histological signs of infection were observed in 26 out of 64 infected larvae, first on day 3 p.i. Of these 26 larvae, 23 were already dead at the time of collection and showed macroscopic changes. The histological findings indicate a fulminant infection process from the germination of the spores in the intestinal lumen to the death of the larva. This study's results are crucial for an improved understanding of the pathogenesis of chalkbrood disease. Chalkbrood is a mycological brood disease of the Western honey bee (Apis mellifera), caused by the fungus Ascosphaera apis. The aim of this study was the investigation of the pathology of artificially reared Apis mellifera larvae, experimentally infected with A. apis spores (1.0 × 103 spores/larva). Non-infected larvae served as control. Five living larvae and every dead larva were collected daily (day 1–7 p.i.). All larvae were macroscopically measured, photographed, formalin-fixed, and histologically processed (hematoxylin-eosin stain, Grocott silvering). Histological sections were digitized, and the size of the larvae was measured (mouth-after length, area) and statistically analyzed. Twenty-six larvae from the collected larvae (n = 64; 23 dead, 3 alive) showed histological signs of infection from 3 d p.i. onwards. The dead larvae showed macroscopically white/brown deposits, indistinct segmentation, and a lack of body elongation. Infected larvae were significantly smaller than the controls on days 3 p.i. (p < 0.05), 4 p.i. (p < 0.001), and 6 p.i. (p < 0.05). The early time of death, the low number of transitional stages, and the strong penetration of the larval carcass with fungal mycelium indicate a rapid and fulminant infection process, which is probably relevant for spreading the disease within the colony. [ABSTRACT FROM AUTHOR]

Simple Summary: Mammary carcinomas have been diagnosed with increasing frequency in pet rabbits. Prognostic biomarkers are limited, and the only available treatment is surgical excision. Additional treatment options are needed, e.g., for animals in which metastases to internal organs preclude complete tumor removal. Human breast cancer may express the immunosuppressive enzyme indoleamine 2,3-dioxygenase 1 (IDO1), that represents a prognostic biomarker and a possible therapeutic target. Since previous studies revealed similarities between human breast cancer and pet rabbit mammary carcinomas, this study investigated IDO1 immunostaining in 96 mammary carcinomas of 96 pet rabbits with an average age of 5.5 years. All rabbits with reported sex were female. Variable percentages of IDO1-positive tumor cells were detected in 90 (94%) carcinomas. Furthermore, IDO1 immunostaining was observed in the secretory epithelial cells of the adjacent non-neoplastic mammary tissue. This study provides further information on the molecular features of mammary carcinomas in rabbits. It also shows similarities in IDO1 expression between rabbit mammary carcinomas and human breast cancer. These findings can have a mutual benefit. They could lead the development of novel treatment options for rabbits with mammary carcinomas. In addition, they further support the value of rabbits with mammary carcinomas for breast cancer research. For mammary carcinomas in pet rabbits, prognostic biomarkers are poorly defined, and treatment is limited to surgical excision. Additional treatment options are needed for rabbit patients for which surgery is not a suitable option. In human breast cancer, the immunosuppressive enzyme indoleamine 2,3-dioxygenase 1 (IDO1) represents a prognostic biomarker and possible therapeutic target. This retrospective immunohistochemical study examined IDO1 in 96 pet rabbit mammary carcinomas with known mitotic count, hormone receptor status, and percentage of stromal tumor infiltrating lymphocytes (TILs). Tumors were obtained from 96 pet rabbits with an average of 5.5 years. All rabbits with reported sex (n = 88) were female or female-spayed. Of the carcinomas, 94% expressed IDO1, and 86% had sparse TILs consistent with cold tumors. Statistically significant correlations existed between a higher percentage of IDO1-positive tumor cells, lower mitotic counts, and increased estrogen receptor expression. The threshold for significance was IDO1 staining in >10% of tumor cells. These results lead to the assumption that IDO1 expression contributes to tumorigenesis and may represent a prognostic biomarker and possible therapeutic target also in pet rabbit mammary carcinomas. They also support the value of rabbits for breast cancer research. [ABSTRACT FROM AUTHOR]

Canine urothelial carcinoma (UC) and prostate carcinoma (PC) frequently exhibit the BRAFV595E mutation, akin to the BRAFV600E mutation common in various human cancers. Since the initial discovery of the BRAF mutation in canine cancers in 2015, PCR has been the standard method for its detection in both liquid and tissue biopsies. Considering the similarity between the canine BRAFV595E and human BRAFV600E mutations, we hypothesized that immunohistochemistry (IHC) using a BRAFV600E‐specific antibody could effectively identify the canine mutant BRAFV595E protein. We tested 122 canine UC (bladder n = 108, urethra n = 14), 21 PC, and benign tissue using IHC and performed digital droplet PCR (ddPCR) on all 122 UC and on 14 IHC positive PC cases. The results from ddPCR and IHC were concordant in 99% (135/136) of the tumours. Using IHC, BRAFV595E was detected in 72/122 (59%) UC and 14/21 (65%) PC. Staining of all benign bladder and prostate tissues was negative. If present, mutant BRAF staining was homogenous, with rare intratumour heterogeneity in three (4%) cases of UC. Additionally, the BRAFV595E mutation was more prevalent in tumours with urothelial morphology, and less common in glandular PC or UC with divergent differentiation. This study establishes that BRAFV600‐specific IHC is a reliable and accurate method for detecting the mutant BRAFV595E protein in canine UC and PC. Moreover, the use of IHC, especially with tissue microarrays, provides a cost‐efficient test for large‐scale screening of canine cancers for the presence of BRAF mutations. This advancement paves the way for further research to define the prognostic and predictive role of this tumour marker in dogs and use IHC to stratify dogs for the treatment with BRAF inhibitors. [ABSTRACT FROM AUTHOR]

Simple Summary: Molecular biomarkers in cancer are measurable genomic alterations that can indicate the risk of developing neoplasia, the presence of neoplastic cells, patient outcome, and/or a likely response to therapy. This review discusses the different uses of molecular biomarkers in the veterinary clinic and how their presence can be determined. In particular, we showcase which genomic alterations are currently used to date as molecular biomarkers in the clinic and for what purposes. We also look at biomarkers that are currently being developed and show promise for clinical use. Finally, we consider the important factors that allow a molecular biomarker to move from the research laboratory to the clinic. This review aims to enable veterinarians to understand the benefits of molecular biomarkers in delivering precision veterinary care to dogs and cats with cancer. Molecular biomarkers are central to personalised medicine for human cancer patients. It is gaining traction as part of standard veterinary clinical practice for dogs and cats with cancer. Molecular biomarkers can be somatic or germline genomic alterations and can be ascertained from tissues or body fluids using various techniques. This review discusses how these genomic alterations can be determined and the findings used in clinical settings as diagnostic, prognostic, predictive, and screening biomarkers. We showcase the somatic and germline genomic alterations currently available to date for testing dogs and cats in a clinical setting, discussing their utility in each biomarker class. We also look at some emerging molecular biomarkers that are promising for clinical use. Finally, we discuss the hurdles that need to be overcome in going 'bench to bedside', i.e., the translation from discovery of genomic alterations to adoption by veterinary clinicians. As we understand more of the genomics underlying canine and feline tumours, molecular biomarkers will undoubtedly become a mainstay in delivering precision veterinary care to dogs and cats with cancer. [ABSTRACT FROM AUTHOR]

Simple Summary: In schnauzers, a breed predisposition to squamous cell carcinoma of the digit (dSCC) is well known. In terms of the breed, the variants of giant (GSs), standard (SSs), and miniature schnauzers (MSs), as well as the traits of body size and/or genetic background may have an impact on tumour occurrence and prognosis. We, therefore, conducted a retrospective analysis of the pathological findings of 478 digital SCCs from 417 schnauzers (227 GSs, 174 SSs, and 16 MSs). The results showed that MSs were older when digital SCC appeared, and lesions were mostly (90%) nodular. A gender predisposition for male GSs was identified (p < 0.05). In GSs, nodular lesions were larger than in MSs and SSs. In all breed variants, SCC was mostly diagnosed at the forelimbs, especially at digits 1, 2, and 5. In contrast, at the hindlimbs, the affected toes differed between GSs and SSs. Multiple dSCCs were more common in SSs than in GSs. If dSCC was the cause of death, the survival time was shorter than in dogs dying from other diseases. Metastases occurred in 20% of the cases and led to a significantly shorter survival time in GSs and SSs. The results showed various differences in the dSCC depending on the size variant of the schnauzer. In schnauzers, a breed predisposition to squamous cell carcinoma of the digit (dSCC) is well known. The aim of this study was to compare the clinical and macroscopic findings of dSCCs in giant (GSs), standard (SSs), and miniature schnauzers (MSs). Methods: Pathology reports of 478 dSCCs from 417 schnauzers (227 GSs, 174 SSs, and 16 MSs) were retrospectively evaluated. Results: The MSs were older than the SSs and GSs (p ≤ 0.01). The male GSs were predisposed to dSCC (p < 0.05). In the GSs, the nodular dSCCs were larger than in the MSs (p ≤ 0.05) and SSs (p ≤ 0.001). The digital SCCs were mostly diagnosed at the forelimbs, especially at digits 1, 2, and 5. At the hindlimbs, the affected toes differed between the GSs and SSs. Multiple dSCCs were more common in SSs than in GSs (p = 0.003). If dSCC was the cause of death, the survival time was shorter than in dogs dying from other diseases (p = 0.004). Metastases occurred in 20% of the cases and led to a significantly shorter survival time in both the GSs and SSs (p < 0.001). Conclusions: The results showed various differences in the dSCC depending on the size variant of the schnauzer. [ABSTRACT FROM AUTHOR]

Urothelial carcinoma (UC) is the most common tumour of the canine urinary bladder. Recently, BRAF mutation testing emerged as a diagnostic option, but its prognostic significance is unknown. This study investigates the relationship between BRAF (variant V595E) mutation status and overall survival in UC‐bearing dogs. Seventy‐nine patients histologically diagnosed with UC of the bladder and/or urethra between 2006 and 2019 were included in this retrospective single‐centre‐study. Treatment consisted of meloxicam (n = 39, group 1 'Melox'), mitoxantrone and meloxicam (+/− followed by metronomic chlorambucil; n = 23, group 2 'Chemo') or partial cystectomy followed by meloxicam +/− mitoxantrone (n = 17, group 3 'Sx'). Survival was significantly influenced by treatment (p =.0002) and tumour location (p <.001) in both uni‐ and multivariable analyses. BRAF mutation was identified in 51 tumours (=64.6%) and had no statistically significant influence on overall survival: MST for BRAF‐negative patients 359 versus 214 days for BRAF‐positive dogs (p =.055). However, in BRAF‐positive dogs, survival depended significantly on type of treatment in univariable analysis: MSTs for groups 1–3 were 151, 244 and 853 days, respectively (p =.006); In BRAF‐positive group 2 ('Chemo')‐patients, adjuvant metronomic chlorambucil after mitoxantrone more than doubled MST compared to patients receiving mitoxantrone alone (588 vs. 216 days; p =.030). In contrast, MSTs were not significantly different in BRAF‐negative patients among the three treatment groups (p =.069). Multivariate analysis of these data was not possible due to group size limitations. This study identified tumour location and treatment type, but not BRAF mutation status, as independent prognostic factors for overall survival. [ABSTRACT FROM AUTHOR]