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In this study, a multiplex PCR method was developed for the detection of four diarrhea-associated viruses of canines, including canine bocavirus (CBoV), canine circovirus (CCV), torque teno canis virus (TTCV), and canine kobuvirus (CKV). Four pairs of compatible primers, one specific for each virus, were designed based on conserved sequences. After optimization of parameters such as primer concentration and annealing temperature in single and multiple amplifications, four specific fragments were amplified simultaneously with high sensitivity and specificity in one PCR reaction. The fragments amplified were 165 bp (CBoV), 345 bp (CCV), 506 bp (TTCV), and 666 bp (CKV) in length. The sensitivity of this one-step multiplex PCR is about 10 times lower than that of regular singleplex PCR. There was no cross-reaction with the canine pathogens canine parvovirus (CPV), canine distemper virus (CDV), or canine coronavirus (CCoV). Testing of canine fecal samples from China using the multiplex PCR assay revealed the presence of CBoV, CCV, TTCV, and CKV in 10.1%, 6.2%, 2.8%, and 1.7% of the samples, respectively. The results of multiplex PCR agreed with the singleplex PCR results with a coincidence rate of 100%. In addition, the complete genome sequences of the viruses in three CKV-positive samples were determined and found to be 95.7 - 96.6% identical to the reference strain US-PC0082 and genetically more distant from other animal kobuvirus. The multiplex PCR method established in this study is convenient, with high specificity and sensitivity, which will be helpful for the rapid differential diagnosis of CBoV, CCV, TTCV, and CKV infections., Competing Interests: Declarations Ethical approval All animal experiments conformed to the guidelines and regulatory requirements established by the Animal Care and Use Committee of Qingdao Agricultural University, Shandong Province, China. The owners’ consent was obtained to collect samples from diarrheic dogs. Conflict of interest All authors declare no conflict of interest., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Background: Microfilaria (MF) testing is an essential part of canine heartworm diagnostics, and it is recommended by the American Heartworm Society that a MF test be performed in tandem with antigen testing on every dog, every year, regardless of prevention status or history. There are a variety of methods that can be used to detect MF in canine whole blood; however, these methods widely vary in their sensitivities as well as practical factors, including time investment and cost. Additionally, some MF tests offer the advantage of being quantitative or allowing for morphological or molecular species identification, while other tests should only be used qualitatively., Methods: The purpose of this study is to evaluate the quantitative and qualitative performance of MF tests, including the 20 μL count, wet mount, 9 μL and 40 μL hematocrit tubes, thin smear, thick smear, modified Knott test (MKT), and conventional polymerase chain reaction (PCR)., Results: Qualitatively, there was little difference in the performance of the 20 μL count, wet mount, MKT, and PCR. The MKT and PCR are the optimal MF tests, as these perform most reliably for detecting positives even when the MF per milliliter is relatively low, and in most cases, these two methods also allow for species-level confirmation of the identity. However, PCR tends to be a very costly test, and both PCR and MKT require a greater degree of expertise and time investment to perform than other tests. Even the lowest performance tests, including the thin smear and hematocrit tube methods, can reliably detect MF at very high burdens; although, caution should be advised when using low reliability methods, since there is a greater likelihood of failing to identify MF-positive dogs., Conclusions: Microfilaria (MF) testing is an essential part of heartworm diagnosis and screening in dogs, and test selection should balance practical factors such as cost and time investment with the patient's risk of infection based on prevention status and history, clinical signs, and antigen testing results. This approach to MF testing will help minimize cost while avoiding failure to detect MF in infected dogs, especially when MF burden is low., Competing Interests: Declarations Ethics approval and consent to participate The dogs utilized for blood collection were managed according to protocol A2022 04-009, which had prior approval by the University of Georgia Institutional Animal Care and Use Committee. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Canine parvovirus (CPV) can cause high morbidity and mortality rates in puppies, posing a significant threat to both pet dogs and the breeding industry. Rapid, accurate, and convenient detection methods are important for the early intervention and treatment of canine parvovirus. In this study, we propose a visual CPV detection system called nucleic acid mismatch enzyme digestion (NMED). This system combines loop-mediated isothermal amplification (LAMP), endonuclease for gene mismatch detection, and colloidal gold lateral chromatography. We demonstrated that NMED can induce the binding of the amplicon from the sample to the specific labeling probe, which in turn triggers digestion by the endonuclease. The sensitivity and visual visibility of LAMP were increased by combining endonuclease and colloidal gold lateral chromatography assisted by a simple temperature-controlled device. The sensitivity of the NMED assay was 1 copy/μL, which was consistent with quantitative PCR (qPCR). The method was validated with 20 clinical samples that potentially had CPV infection; 15 positive samples and 5 negative samples were evaluated; and the detection accuracy was consistent with that of qPCR. As a rapid, accurate, and convenient molecular diagnostic method, NMED has great potential for application in the field of pathogenic microorganism detection., Importance: The NMED method has been established in the laboratory and used for CPV detection. The method has several advantages, including simple sampling, high sensitivity, intuitive results, and no requirement for expensive equipment. The establishment of this method has commercial potential and offers a novel approach and concept for the future development of clinical detection of pathogenic microorganisms., Competing Interests: The authors declare no conflict of interest.

An 8-year-old spayed female Maltese dog was referred with a 4-month history of multifocal pruritic erosive or ulcerative lesions on the head, ventral neck, chest, and abdomen; and chronic otitis externa. Biopsy revealed cytotoxic dermatitis and apoptotic keratinocytes with occasional lymphocytic satellitosis. Bacterial culture and antimicrobial susceptibility tests revealed multidrug-resistant Pseudomonas aeruginosa infection of the skin and ears. The skin lesions regressed after treatment of P. aeruginosa infection with ciprofloxacin. Based on the skin lesions, histological characteristics, and response to therapy, the dog was presumed to have erythema multiforme induced by P. aeruginosa infection. This case report emphasizes that P. aeruginosa infection could be a trigger of erythema multiforme in dogs. Key clinical message: Erythema multiforme caused by infection is rare and poorly characterized in the veterinary literature. This case report describes the clinical characteristics of, diagnostic approach to, and treatment of erythema multiforme caused by Pseudomonas aeruginosa ., (Copyright and/or publishing rights held by the Canadian Veterinary Medical Association.)

An approximately 3 yr old female Chihuahua was presented for evaluation of polyuria and polydipsia noted by the owner after adoption. Serum biochemistry and urinalysis revealed hyponatremia, hypochloremia, hypo-osmolality, and normal fractional excretion of sodium. Serum antidiuretic hormone concentration was 5.54 pg/mL and serum copeptin concentration was 135.40 pg/mL. The antidiuretic hormone value was above the reported range for healthy dogs and was considered inappropriate in light of serum osmolality. Idiopathic syndrome of inappropriate antidiuretic hormone secretion was considered the most likely diagnosis after excluding other potential causes, such as diuretic use, renal disease, adrenal disease, and thyroid dysfunction. The dog remains clinically well 2 yr after diagnosis without specific treatment other than minimal dietary salt supplementation. This case is unique because of the absence of systemic or neurologic signs, good long-term outcome without treatment, and use of novel diagnostic testing., (© 2024 by American Animal Hospital Association.)

A 12 years-old, male indigenous dog was presented to Shahedul Alam Qaderi Teaching Veterinary Hospital (SAQTVH) in Chattogram, Bangladesh with a large tumour on its right front leg. The histopathological findings of the tumour were highly suggestive of an alveolar rhabdomyosarcoma (ARMS). The tumour was surgically removed from the dog. While there had been no recurrence for 5 months, the patient died 5 months post-operatively. The cause of the death and presence of metastasis could not be confirmed as follow-up investigations were declined by the owner. A histopathological study showed poorly differentiated neoplastic mononuclear cells forming nests in an alveolar pattern. Despite that the limited resources prevented myogenin staining for diagnostic confirmation, the well-contained, defined appearance of the tumour supports the diagnosis of ARMS. This report highlights the challenges in diagnosing and treating ARMS in indigenous dogs and emphasizes the need for further research and diagnostic markers to improve detection and management., (© 2024 The Author(s). Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

A 10-year-old female neutered Labrador Retriever presented with epistaxis, discoloration and crusting of the nose and a necrotic lesion on the lip. Bloodwork revealed pancytopenia, azotemia, hypoalbuminemia and hyperglobulinemia. Aggregates of amorphous basophilic material were seen in a room-temperature blood smear which were not present in the sample after warming to 37°C, and grossly a cryoprecipitate was noted in the patient's serum at 4°C. This was interpreted as cryoglobulin. Computed tomography showed multiple heterogeneous lesions in the spleen. Cytology of the splenic lesions revealed marked plasma cell infiltration, consistent with neoplasia. Bone marrow aspiration revealed an increased proportion of plasma cells (approximately 38% of the total cells). Serum protein electrophoresis showed a monoclonal spike in the gamma globulin region. A diagnosis of multiple myeloma associated with cryoglobulinemia was made. The patient received palliative care with prednisolone while the owner was considering chemotherapy. However, she rapidly deteriorated and was euthanized. The combination of cryoglobulin precipitation and hyperviscosity syndrome was considered responsible for the patient's original symptoms. Cryoglobulinemia is an extremely rare phenomenon that is often associated with lymphoproliferative disorders. This report describes its association with multiple myeloma in a dog presenting with atypical initial signs., (© 2024 The Author(s). Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Lymphoma ranks among the most prevalent neoplasms in veterinary oncology, frequently diagnosed in dogs, particularly in its multicentric form. While histopathology plays a crucial role in lymphoma diagnosis, prognosis and prediction of biological behavior, minimally invasive diagnostic methods are increasingly emerging as viable alternatives. This study aims to assess and compare various minimally invasive diagnostic techniques for multicentric lymphomas in dogs. A total of 38 dogs, encompassing various sexes, ages, and breeds, with clinical suspicion of multicentric lymphoma, was included in the study. Fine needle aspiration was employed to collect samples from lymph nodes, which were subsequently used for cytology, cell block preparation, PCR for antigen receptor rearrangement (PARR), and immunocytochemistry. Among the animals evaluated, 31 dogs received a cytological diagnosis of lymphoma, while 7 showed findings suggestive of lymphoma or lymphadenitis. Immunocytochemistry on cytological smears yielded inconclusive results in 50 % of cases, with 44.74 % diagnosed with B-cell lymphoma and 5.26 % with T-cell lymphoma. Cell block analysis identified lymphoma in 30 dogs and suggested lymphoma or a round cell neoplasm in 8 cases. Cell block immunocytochemistry confirmed lymphoma in 35 dogs, comprising 80 % B-cell and 20 % T-cell lymphomas. PARR revealed monoclonal rearrangement/clonality in 33 cases, with 84.85 % of these being B-cell and 15.15 % T-cell lymphomas. This study underscores the precision of minimally invasive techniques in diagnosing and characterizing multicentric lymphoma in dogs, reaffirming their significance in veterinary clinical practice., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

MRI is widely used for the detection and characterization of brain lesions. There is a growing interest in the potential benefits of quantitative MRI (qMRI) in veterinary brain lesion diagnosis. Yet, the use of data processing tools in the veterinary field is not as democratized as for the diagnosis of human brain pathologies. Several reviews have addressed the characterization of brain lesions in cats and dogs. None of them is specifically focused on quantitative MRI data processing techniques for the diagnosis of brain lesions in the veterinary field. This paper aims to provide an overview of the evolution of qMRI on cats and dogs both in the clinical and preclinical fields. We analyze the achievements in the field as well as the remaining challenges in the diffusion of data processing tools for veterinary brain lesions characterization., (© 2024 HawkCell and The Author(s). Veterinary Radiology & Ultrasound published by Wiley Periodicals LLC on behalf of American College of Veterinary Radiology.)

Objective: To compare the sensitivity and specificity of pH with multichannel intraluminal impedance (pH-MII), pH-metry (pH) alone and MII alone to direct observation of GOR by endoscopy in anaesthetized dogs., Study Design: A prospective comparative trial in a live canine model., Animals: A group of 35 (22 females, 13 males) dogs of various breeds. The mean (range) body weight and age were 31.9 (14-40) kg and 5.6 (0.75-12) years, respectively., Methods: All dogs were premedicated with medetomidine and morphine, anaesthesia was induced with propofol and maintained on isoflurane in oxygen. A monitoring assembly consisting of an endoscopy camera, endotracheal tube and a disposable flexible pH-MII catheter was used to measure oesophageal pH, MII and directly visualize reflux. Visual reflux score was (0-3) and pH was recorded on a data capture sheet. Reflux was considered to have occurred whenever oesophageal pH was < 4.0 or > 7.5, device software analysing MII data detected fluid or a visual reflux score of 2 or 3 were assigned. Receiver operator curves (ROC) analysis was used to determine sensitivity and specificity for each monitoring method to detect GOR., Results: Endoscopy identified GOR in 20 dogs (57%), pH-MII in 19 dogs (54%), pH alone in 13 dogs (37%) and MII alone in 12 dogs (24%). ROC analysis showed fair accuracy for pH-MII and pH alone, whereas MII demonstrated low accuracy., Conclusions and Clinical Relevance: In conclusion, pH-MII is a reliable method for detecting GOR and emerges as a promising tool for future research. Endoscopy is reliable and provides the ability to subjectively quantify the volume of reflux; however, it lacks the ability to discern the pH of refluxate. pH alone misses reflux events with intermediate pH (4.1-7.4). Incorporation of impedance addresses some of the limitations associated with pH alone and enhances diagnostic accuracy., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Melanoma is the most common malignant oral tumor in dogs. It frequently presents a diagnostic challenge as many melanomas lack or contain scant melanin and may have a variable microscopic phenotype. Previous studies evaluating immunohistochemical markers for diagnosing melanoma have shown limited sensitivity and/or specificity for S-100, PNL2, melan A, TRP-1, TRP-2, and HMB-45. Sry-related HMG-box gene 10 (SOX-10) is a transcription factor associated with melanocytic, peripheral neural crest, and peripheral nervous system development. In humans, SOX-10 expression has been demonstrated in melanoma, breast carcinoma, glioma, and schwannoma, but has only recently been explored in veterinary species. In this study, 198 tumors comprised of 147 melanocytic neoplasms and 51 non-melanocytic neoplasms were evaluated by immunohistochemistry using a tissue microarray for SOX-10, PNL2, melan A, TRP-1, and TRP-2 expressions. The SOX-10 had the highest diagnostic sensitivity (96.7%) in melanomas. In addition, SOX-10 had the highest percentage (91.5%; 130/142) of melanomas label at least 75% of neoplastic cells. Of the 51 selected non-melanocytic tumors examined, SOX-10 labeling was observed in mammary carcinomas (6/6), gliomas (4/4), and oral soft tissue sarcomas (4/18). Of the 41 non-melanocytic oral neoplasms evaluated, SOX-10 had a specificity of 92.7%. Therefore, SOX-10 represents a useful immunohistochemical screening marker for the diagnosis of canine melanoma given its extremely high sensitivity and robust labeling intensity. The SOX-10 may have utility in diagnosing some non-melanocytic neoplasms in the dog, although this requires further investigation., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

A 10-y-old spayed female Cavalier King Charles Spaniel dog was presented to the Veterinary Teaching Hospital because of recurrent chronic abscesses on the distal pelvic limbs, fever, lethargy, lameness of unknown etiology, and chronic pancreatitis. Sterile nodular panniculitis was diagnosed after an extensive workup, and the dog initially responded to immunosuppressive therapy, but relapse and spread of cutaneous lesions and acute lameness occurred after 11 mo, and euthanasia was elected. Postmortem examination confirmed hyalinizing pancreatic adenocarcinoma with pancreatitis, panniculitis, polyarthritis (PPP), and osteomyelitis. Histopathology and bacterial and fungal cultures were supportive of a sterile process, specifically the PPP syndrome, which is a rare, potentially life-threatening, systemic manifestation of pancreatic disease in both people and animals. To our knowledge, a clinicopathologic description of a hyalinizing pancreatic adenocarcinoma associated with this rare syndrome has not been reported previously in a dog., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Histologic diagnosis of less well-differentiated cases of canine extramedullary plasmacytomas (CEMPs) may require immunohistochemical confirmation to discriminate these tumors from other round cells tumors including lymphoma, cutaneous histiocytoma, and amelanotic melanomas. CEMPs are characterized by widespread immunoreactivity for multiple myeloma 1 (MUM1) antigen and λ light chains, while the melanocytic marker melan-A has been reported to yield negative results. Here, 33 randomly selected CEMPs, 20 melanocytomas, and 20 malignant melanomas were immunohistochemically tested for MUM1, melan-A, and PNL2. In addition, CEMPs were examined for PAX5, E-cadherin, CD3, CD18, CD20, S100, as well as λ and κ light chain immunoreactivity. All CEMPs were characterized by labeling for MUM1 and λ light chain, as well as variable immunopositivity for the remaining antibodies. Notably, 13 cases of CEMPs (39.4%) exhibited immunolabeling for melan-A. Melanocytic tumors immunolabeled for melan-A (40/40; 100%) and PNL2 (34/40; 85%). An unexpected cytoplasmic immunoreactivity for MUM1 was observed in 2 melanocytic tumors. Summarized, MUM1 or melan-A immunomarkers alone are not sufficient to differentiate between CEMPs and amelanotic melanomas and should be part of a larger immunopanel including λ light chain, CD20, and PNL2., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Neuromuscular and vascular hamartoma (NMVH) is an infrequent gastrointestinal lesion described in human and veterinary medical literature. The histologic features of this entity are haphazardly arranged fascicles of smooth muscle, nerve fibers, scattered ganglion cells, and hemangiomatous blood vessels. Here we describe 2 putative cases of NMVH in a 1.7-y-old, intact female Anatolian mixed-breed dog and a 4-mo-old intact male Akita dog. Both animals had gastrointestinal clinical signs, including hematochezia, and on exploratory laparotomy, intussusception was confirmed. Histologic examination confirmed NMVH within the cecal wall in both cases using a panel of immunohistochemical (IHC) markers for vascular structures (CD31), smooth muscle (alpha-smooth muscle actin [α-SMA]), and nerves (glial fibrillary acidic protein [GFAP] and S100). The complete surgical excision of the lesion in both animals was considered curative without persistent clinical signs 14 mo and 12 mo, respectively, after surgery., Competing Interests: Declaration of conflicting interestsTatiane Terumi Negrão Watanabe and Linda Huang are employed by Antech Diagnostics, Mars Petcare Science & Diagnostics. The remaining authors declared no potential conflicts of interest concerning the research, authorship, and/or publication of this article.

High survivin expression has been correlated with poor outcomes in several canine tumors but not in soft tissue tumors (STTs). Survivin is a target gene of the Wnt/β-catenin pathway, which is involved in human STT oncogenesis. Immunohistochemistry for survivin, β-catenin, and Ki-67 was performed on 41 canine perivascular wall tumors (cPWTs), and statistical associations of protein expression and histopathologic and clinical variables with clinical outcomes were investigated. Immunohistochemically, there was nuclear positivity (0.9%-12.2% of tumor cells) for survivin in 41/41 (100%), cytoplasmic positivity (0 to > 75% of tumor cells) for survivin in 31/41 (76%), nuclear positivity (2.9%-67.2% of tumor cells) for β-catenin in 24/41 (59%), and cytoplasmic positivity (0% to > 75% of tumor cells) for β-catenin in 23/41 (56%) of cPWTs. All tumors expressed nuclear Ki-67 (2.2%-23.5%). In univariate analysis and multivariate analysis (UA and MA, respectively), every 1% increase of nuclear survivin was associated with an increase of the instantaneous death risk by a factor of 1.15 [hazard ratio (HR) = 1.15; P = .007]. Higher nuclear survivin was associated with grade II/III neoplasms ( P = .043). Expression of cytoplasmic survivin, nuclear and cytoplasmic β-catenin, and nuclear Ki-67 were not significantly associated with prognosis in UA nor MA. Tumor size was a significant prognostic factor for local recurrence in UA [subdistribution HR (SDHR) = 1.19; P = .02] and for reduced overall survival time in MA. According to UA and MA, a unitary increase of mitotic count was associated with an increase of the instantaneous death risk by a factor of 1.05 (HR = 1.05; P = .014). Nuclear survivin, mitotic count, and tumor size seem to be potential prognostic factors for cPWTs. In addition, survivin and β-catenin may represent promising therapeutic targets for cPWTs., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

A 10-y-old spayed female Staffordshire Terrier dog was evaluated because of a cutaneous left ear base mass. Cytology revealed sheets of cells with anisocytosis and anisokaryosis, round-to-oval or plasmacytoid cytoplasm, and round, central, or eccentric nuclei; binucleate cells were present. Cytologic findings were consistent with a round cell tumor (plasmacytoma or agranular mast cell tumor), amelanotic melanoma, or anaplastic carcinoma. Histologically, neoplastic cells were polygonal to elongate, with round-to-oval nuclei and prominent nucleoli and arranged in sheets and nests on a fibrovascular stroma. Neoplastic cells with plasmacytoid morphology (round, glassy, eosinophilic cytoplasm with eccentric nuclei) were present in ~30% of the neoplasm. There were 18 mitoses in 2.37 mm 2 (10 FN22/40× fields). Neoplastic cells had cytoplasmic immunolabeling for melan A and PNL2 and no immunolabeling for AE3/1 and MUM1, consistent with a dermal melanoma with plasmacytoid differentiation. The patient was re-evaluated ~1 mo after the first biopsy because of local recurrence of the original mass and new masses on the interscapular area and right elbow; these neoplasms were histologically identical to the original submission, plus scattered neoplastic cells in the new masses contained brown cytoplasmic pigment. The dog was euthanized because of swelling and hemorrhage of the tumors and right pelvic limb lameness. Our findings were consistent with a dermal melanoma with plasmacytoid features that were similar to human plasmacytoid melanoma, a rare variant of human melanoma that is diagnostically challenging as it may mimic a plasmacytoma., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

A 6-month-old intact male Dalmatian mix puppy was presented for the evaluation of left maxillary swelling due to a suspected cyst and an unerupted left maxillary canine tooth. Removal of the unerupted left maxillary canine tooth (204) and enucleation of the cyst was performed, followed by histological analysis, which identified the maxillary swelling to be a cystic ameloblastic fibroma. Ameloblastic fibromas are rare in companion animals, and to the best of the authors' knowledge, this is the first cystic variant reported in dogs. The clinical, radiographic, cone beam computed tomography, and histological findings of this case are discussed and compared with the findings of previously documented human and domestic animal cases., Competing Interests: Declaration of Conflicting InterestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Histologic grading of canine cutaneous mast cell tumors (cMCTs) has prognostic and therapeutic implications, yet validation for subcutaneous MCTs (scMCTs) is lacking. For scMCTs with or without dermal invasion, determining their biological behavior remains poorly standardized and sometimes sparks controversy. This prospective study aimed to assess the prognostic utility of the 2-tier histologic grading system in MCTs with different growth models (GMs) and explore the prognostic impact of the GM itself. We assessed 6 histologic GM categories: solely cMCT (C-SC 0 ), cMCT with superficial (C-SC 1 ) or deep subcutaneous (C-SC 2 ) involvement, solely scMCT (SC-C 0 ), and scMCT with deep (SC-C 1 ) or superficial (SC-C 2 ) infiltration of the dermis. Ninety-one MCTs from 76 dogs undergoing excision and regional/sentinel lymphadenectomy were examined. GM classification identified 11 (12%) C-SC 0 tumors, 12 (13%) C-SC 1 , 15 (16%) C-SC 2 , 21 (23%) SC-C 0 , 15 (16%) SC-C 1 , and 17 (19%) SC-C 2 . Mitotic count, 2-tier grade, nodal involvement, surgical margins, and outcome were stratified according to GM. scMCTs lacking dermal invasion, historically associated with a benign clinical course, had a poor prognosis in 10% of cases. cMCTs exhibiting deep subcutaneous involvement included the largest percentage of high-grade tumors (33%), had the highest occurrence of overt nodal metastases (33%), and had the lowest 1-year survival rate (86%). Histologic grade was confirmed as a relevant prognostic factor, surpassing nodal involvement and histologic margin status. The 2-tier histologic grading enabled the identification of all MCTs with aggressive biological behavior, regardless of their cutaneous or subcutaneous location., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Aims: To assess agreement of bacterial culture results from samples taken from nasal discharge, the nasal cavity and nasal biopsy from dogs and cats with nasal disease., Methods: Nineteen dogs and 21 cats with different nasal diseases (chronic rhinitis, n = 30; neoplasia, n = 7; sinonasal aspergillosis, n = 3) were prospectively enrolled in the study. Nasal swabs were taken bilaterally from nasal discharge at the nares, the nasal cavity, and one nasal mucosal biopsy per side. All samples were subjected to aerobic bacterial culture. Kappa statistics were used to evaluate agreement for the most prevalent bacterial species between sampling sites., Results: A positive culture result for at least one bacterial species was detected in 80% of samples from nasal discharge/nares, 92% of nasal cavity samples, and 75% of biopsy samples. The mean agreement between the three sampling sites for positive vs. negative culture results was never greater than moderate and the precision of the estimates of agreement varied widely.The most frequently isolated bacterial species in dogs were Staphylococcus pseudintermedius , Staphylococcus spp. and Streptococcus spp. In cats, Pasteurella spp. and Staphylococcus felis were the bacterial species cultured most frequently.For the most prevalent cultured species, Staphylococcus spp., mean agreement between sites was never greater than fair and the precision again varied widely., Conclusion: This study indicates that bacterial culture results in feline and canine nasal disease are site-specific and there was no evidence from this study for consistency between sites within a patient for many bacterial species. Consequently, if bacterial culture results from nasal swabs are used to guide therapeutic antimicrobial choice, different treatments may be selected depending on the site of culture. As a consequence, there is no evidence from this study that nasal bacterial cultures should be recommended as a routine diagnostic measure.

Synovial myxoma, a rare joint tumor in dogs, has traditionally been considered benign, acknowledging that local invasion into regional tissues including bone may be present. Given the diagnostic challenges in distinguishing synovial myxoma from other joint lesions through clinical features and diagnostic imaging, definitive diagnosis relies on characteristic gross and histologic features. Within the inner surface of the joint capsule, synovial myxomas form nodules of stellate-to-spindle cells within abundant myxomatous matrix. We present here 2 cases of synovial myxoma with metastasis to regional lymph nodes and compare these 2 cases to 3 cases without evidence of lymph node metastasis. Aside from lymphovascular invasion in one case with metastasis, there were no overt histologic features of the primary tumor to suggest aggressive biologic behavior. The finding of lymph node metastasis warrants reconsideration of the term "synovial myxoma" for this neoplasm. We suggest the term "synovial myxosarcoma," considering that histologic features of the primary tumor do not predict biologic behavior. Our case series highlights the importance of lymph node sampling in suspected synovial myxosarcoma cases as well as thorough histologic examination, emphasizing careful evaluation for lymphovascular invasion., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

A 2-year-old male neutered Standard Poodle weighing 17.9 kg was presented to their primary care veterinarian for enlarged bilateral submandibular swellings following an interdog altercation sustained in the previous weeks. Cytology performed following fine-needle aspirates of the regions of swelling was inconclusive, and the patient was treated empirically with Clavaseptin. Despite treatment, the submandibular swellings continued to enlarge, and right-sided intermittent epistaxis was reported. On biochemical profile, there was mild hypercalcemia and mild hyperglobulinemia. The computed tomography (CT) findings were indicative of severe multifocal sialadenitis with severe regional cellulitis and inflammatory lymphadenopathy. Histopathology and cytology results described mixed inflammation of the salivary gland. Methenamine silver staining and Fite's acid-fast staining were negative. Aerobic and anaerobic cultures were negative. Targeted, next-generation DNA sequencing detected no known fungi or bacterial pathogens. These findings were consistent with the diagnosis of severe bilateral mandibular sialadenitis, panniculitis, and lymphadenopathy. The patient was prescribed enrofloxacin, clindamycin, phenobarbital, and prednisolone for 1 month. One week after initiating treatment, the patient had a significant reduction in size of the salivary glands. CT imaging was helpful in the diagnosis of this patient and allowed the clinician to identify which submandibular anatomical structures were abnormal, guiding subsequent diagnostic decisions to provide medical management to resolve the condition., Competing Interests: Declaration of Conflicting InterestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

A 10-year-old golden retriever was presented for the evaluation of exercise intolerance. Physical examination and laboratory tests showed no abnormalities except for mild anaemia. Standard transthoracic echocardiography revealed dilated coronary sinus, raising suspicion of a persistent left cranial vena cava, and the persistency of this anomaly was confirmed by agitated-saline study. Right ventricular echocardiographic parameters did not change significantly, while conventional parameters and global longitudinal strain analysis revealed left ventricular systolic dysfunction. Moreover, post-systolic shortening and early systolic lengthening were detected by two-dimensional speckle tracking echocardiography. This report shows that post-systolic shortening and early systolic lengthening may be useful diagnostic markers indicating regional left ventricular systolic dysfunction. Further studies are needed to elucidate whether persistent left cranial vena cava contributes to left ventricular systolic dysfunction or is an accidental benign finding in dogs., (© 2024 British Small Animal Veterinary Association.)

Objectives: To compare standard tests of haemostasis in hypercoagulable and normocoagulable sick dogs as identified by thromboelastography., Materials and Methods: We compared prothrombin time, activated partial thromboplastin time, D-dimers, packed red blood cell volume and platelet counts in client-owned sick dogs with hypercoagulable (n = 36) and normocoagulable (n = 35) states as identified by thromboelastography maximum amplitude above or within the established reference interval, respectively., Results: Seventy-one dogs were included. Thirty-six dogs were hypercoagulable and 35 were normocoagulable on thromboelastography. Using multivariable linear models, prothrombin time, activated partial thromboplastin time and D-dimers were not associated with hypercoagulability. Platelet counts and packed red blood cell volume were significantly associated with hypercoagulability., Clinical Significance: Prothrombin time, activated partial thromboplastin time and D-dimers were not useful in the identification of hypercoagulability in dogs as defined by thromboelastography., (© 2024 The Author(s). Journal of Small Animal Practice published by John Wiley & Sons Ltd on behalf of British Small Animal Veterinary Association.)

This case study describes the diagnosis and treatment of a deciduous left maxillary canine tooth root that had been displaced into the pulp of the developing permanent left maxillary canine tooth in a 23-month-old female neutered Cavalier King Charles Spaniel. The patient was initially presented for bilateral persistent deciduous maxillary canine teeth extraction. Seventeen months later the permanent left maxillary canine tooth was found to be non-vital and tooth development had ceased prior to apical closure. Radiographs revealed a radio-opaque dentine-like structure and straight line centrally within the pulp of the permanent tooth. The displacement of the deciduous tooth into the developing permanent tooth was confirmed. This unusual potential complication should be considered when performing deciduous teeth extraction., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

A dog presented with a 1-month history of left-sided hemiparesis. MRI showed a focal, 4-cm-long, symmetrical, ovoid, poorly demarcated intramedullary expansion at C6-C7 that was T2-weighted hyperintense, T1-weighted isointense, and noncontrast enhancing. After clinical progression and euthanasia, pathology revealed a neoplasm composed of astrocytes and dysmorphic neurons, consistent with a ganglioglioma. The diagnosis was confirmed with immunohistochemistry and transmission electron microscopy, which demonstrated electron-dense granules in the perikaryon. Gangliogliomas are rare, benign neoplasms that may present as intramedullary spinal cord neoplasia. This is the first report on the clinical presentation, imaging, and pathology of a canine spinal ganglioglioma., (© 2024 American College of Veterinary Radiology.)

Background: Chronic lymphoid leukemia (CLL) is a hematological disorder characterized by the clonal expansion of small mature lymphocytes that accumulate in the blood and bone marrow. CLL can arise from B-, T-, or natural killer cell clones. The cytological evaluation of blood smears is often the simplest and least invasive method for diagnosing lymphoid leukemia. Immunophenotyping is used to further subclassify the type of lymphoid leukemia., Case Presentation: A 15-year-old, 4.4-kg spayed female Shih Tzu was presented to the veterinary medical teaching hospital of Kangwon National University. Despite having a normal appetite and activity level, cervical and inguinal lymph node enlargement was noted on physical examination. Complete blood count revealed severe leukocytosis, severe lymphocytosis, and monocytosis. Splenomegaly, hepatomegaly, and lymph node enlargement were detected on radiographic and ultrasonographic examination. Immunophenotyping was performed using peripheral blood mononuclear cells (PBMCs). The majority of lymphocytes exhibited the following profiles: CD3 - CD79a - (97.5%), CD4 - CD8 - (98.6%), CD21 - CD79a - (98.4%), CD34 - (0.1%), CD45 + (99.6%), major histocompatibility complex class II + (99.5%), and CD14 - (0.5%). Based on the immunophenotyping results, possible differentials considered included the following: the majority of lymphocytes may be natural killer (NK) cell clones, plasma cell clones, or show aberrant expression or loss of CD21 marker due to the neoplastic nature of the cells. Further flow cytometry was performed using antibodies against CD3, CD5, CD94, and granzyme B. The combined results indicated that the predominant lymphocyte subset in the PBMCs was CD3 - CD5 - CD21 - CD94 - granzyme B - . To confirm monoclonality and exclude the aberrant loss of CD markers, a polymerase chain reaction for antigen receptor rearrangement (PARR) assay was conducted. The PARR assay, using DNA from blood and lymph node samples, showed B-cell monoclonality. Immunocytochemistry using PBMCs showed that the plasma cell marker Multiple Myeloma Oncogene 1 (MUM1) was not expressed. Therefore, the diagnosis was confirmed to be B-cell CLL., Conclusion: Immunophenotyping can help subclassify the type of lymphoid leukemia; however, as tumor cells can show aberrant expression or loss of the CD21 marker, combining immunophenotyping with the PARR assay could yield a more accurate diagnosis., (© 2024. The Author(s).)

Domestic dogs (Canis lupus familiaris) serve as valuable translational models for human cancer research due to their biological similarities. Canine mammary tumors (CMTs), frequently diagnosed in female dogs, share various characteristics with human breast cancers. This study investigates the PIK3CA (H1047R) mutation in CMTs using droplet digital PCR (ddPCR) and explores the potential of liquid biopsy for non-invasive detection. We analyzed 80 formalin-fixed, paraffin-embedded (FFPE) CMT tissue samples and compared ddPCR results with next-generation sequencing (NGS) data, achieving high concordance. Plasma and serum samples were also assessed for mutation concordance with tissue results. Our findings indicate a higher frequency of the PIK3CA (H1047R) mutations in benign and grade I malignant CMTs compared to more aggressive malignancies. The ddPCR assay demonstrated high sensitivity and specificity, with plasma testing showing 78.6% sensitivity and 87.5% specificity, and serum testing showing 66.7% sensitivity and 90.0% specificity. These results highlight the viability of liquid biopsy as a minimally invasive method for monitoring PIK3CA mutations in canine patients. The study suggests that liquid biopsy techniques hold significant promise for improving the early detection and monitoring of canine cancers, warranting further research to refine these methods and explore their applications in canine cancer diagnostics and treatment., (© 2024. The Author(s).)

Background: Toxocara canis and Toxocara cati are parasitic nematodes that occur worldwide. As embryonated Toxocara spp. eggs in the environment pose a zoonotic risk, especially for children, optimal diagnostic approaches are necessary for effective disease response and management, including surveillance. However, little is known about the performance of different diagnostic protocols for detecting Toxocara spp. in the faeces of cats and dogs, hampering movement towards an optimal diagnostic process. This study aimed to compare detection methods, including a newly developed sequential sieving protocol (SF-SSV) and a high-throughput multiplex qPCR-based method to facilitate epidemiological studies., Methods: Species-specific Toxocara spp. egg suspensions and canine and feline faecal samples from the field were used to estimate analytical and diagnostic sensitivity of the protocols. The performance of two automated DNA extraction protocols using enzymatic and mechanical lysis were compared by multiplex qPCR, targeting both T. canis and T. cati-specific genomic sequences. All samples were examined by microscopy-based techniques, the sedimentation flotation technique (SF) and a newly developed SF-SSV for the detection, enrichment and purification of parasite eggs. The costs and processing times necessary for all protocols were estimated and compared for both single samples and sets of 100 samples., Results: To detect Toxocara spp. eggs, SF-SSV showed the highest analytical sensitivity and a significantly higher diagnostic sensitivity than the DNA detection methods. Mechanical lysis performed better than enzymatic lysis for automated DNA extraction. In automated DNA extraction, 96-well plates performed better than 24-well plates. DNA detection and microscopy-based parasitological methods showed substantial agreement between the results generated by each method. Microscopy-based techniques required the lowest costs and least hands-on time for a single sample. However, when costs and labour were estimated for a set of 100 samples, the DNA detection protocol using 96-well plates for extraction revealed costs similar to SF-SSV and the fastest processing times., Conclusions: SF-SSV was superior in terms of analytical and diagnostic sensitivity for the detection of Toxocara spp. eggs. For larger sets of samples, multiplex qPCR-based DNA detection represents an alternative to microscopy-based methods, based on the possibility of faster sample processing at similar costs to SF-SSV, and the ability to provide species-specific diagnoses., (© 2024. The Author(s).)

Haemoparasitic infections are frequently observed in dogs from tropical regions, including India. The present investigation combined microscopic blood smear examination and PCR assays to assess the occurrence of canine tick-borne diseases (CTBD) from suspected dogs in and around Hisar, Haryana. Using the Giemsa-stained peripheral thin blood smear examination, 15 (12.5%) of the 120 dogs were infected with CTBD, with 5.8%, 3.3%, 2.5%, and 0.8% dogs testing positive for Hepatozoon canis, Ehrlichia canis, Babesia vogeli, and Babesia gibsoni, respectively. Using the PCR assay, CTBD was found to be 64.16% (77/120) in examined dogs. Of the 77 PCR-positive canines, 56 were infected with a single haemoparasite, while 21 were infected with two or more species. H. canis was the most abundant tick-borne pathogen, representing 35%, followed by E. canis 25.8%, B. vogeli 20%, and B. gibsoni 2.5%. The most common co-infection was with H. canis along with E. canis (7.5%). The PCR assay was proven to be more efficient for detecting haemoparasites in dogs compared to blood smear examinations. The study suggests that canine tick-borne diseases are common in Haryana and recommends using PCR-based molecular tests in addition to conventional microscopic examination to diagnose these infections for effective treatment and management of infected canines., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The cerebral biomarkers, neurofilament light chain (NfL), amyloid-β, tau, and neuron specific enolase (NSE) reflect a wide spectrum of neurological damage in the brain and spinal cord. With this study, we aimed to assess whether these biomarkers hold any potential diagnostic value for the three most common canine neurological diseases. Canines suffering from meningoencephalitis of unknown origin (MUO), brain tumors, and selected non-infectious myelopathies were included. For each diagnosis, we analyzed these biomarkers in the cerebrospinal fluid collected via cranial puncture from the cisterna magna. Elevated levels of CSF tau, NfL, and NSE were observed in MUO, with all three biomarkers being intercorrelated. Tau and NSE were increased while amyloid-β was decreased in dogs suffering from tumors. In contrast, no biomarker changes were observed in dogs with myelopathies. Covariates such as age, sex, or castration had minimal impact. CSF biomarkers may reflect molecular changes related to MUO and tumors, but not to non-infectious myelopathies. The combination of NfL, tau, and NSE may represent useful biomarkers for MUO as they reflect the same pathology and are not influenced by age., (© 2024. The Author(s).)

The enteric protozoan parasites Giardia duodenalis and Cryptosporidium spp. are common cause of diarrhea in pet dogs and cats, affecting primarily young animals. This comparative study evaluates the diagnostic performance of conventional and molecular methods for the detection of G. duodenalis and Cryptosporidium spp. infection in dogs and cats.The compared diagnostic assays included merthiolate-iodine-formalin (MIF) method, lateral flow immunochromatography rapid test (ICT) and real-time PCR; using direct immunofluorescence assay (DFA) as golden standard. The study included the analysis of 328 fecal samples from different dog (n = 225) and cat (n = 103) populations.According to DFA, the overall prevalence of G. duodenalis was 24.4% (80/328, 95% CI: 19.8-29.4), varying from 11.6% (12/103, 95% CI: 6.2-19.5) in cats to 30.2% (68/225, 95% CI: 24.3-36.7) in dogs. The overall prevalence of Cryptosporidium spp. was 4.0% (13/328, 95% CI: 2.1-6.7), varying from 2.9% (3/103, 95% CI: 0.6-8.3) in cats to 4.4% (10/225, 95% CI: 2.1-8.0) in dogs. MIF was only used for the detection of G. duodenalis, which was identified by this method in 22.7% of dogs and 7.8% of cats, respectively. DFA was the most sensitive technique for detecting G. duodenalis in samples from dogs and cats (p-value: < 0.001), followed by real-time PCR. Identification of Cryptosporidium infections was most effectively accomplished by the combination of DFA and PCR technique (p-value: < 0.001). In addition, epidemiological (sex, age, origin) and clinical (fecal consistency) variables were collected to assess their potential associations with an increased likelihood of infection by G. duodenalis and/or Cryptosporidium spp. Breeder dogs were more likely to harbor G. duodenalis infection (p-value: 0.004), whereas female cats were significantly more infected with Cryptosporidium (p-value: 0.003).In conclusion, DFA (alone or in combination with PCR) has been identified as the most accurate and cost-effective method for detecting G. duodenalis and Cryptosporidium spp. in fecal samples from pet dogs and cats. This highlights their importance in both veterinary and clinical settings for enabling prompt treatment and preventing potential transmission to humans., (© 2024. The Author(s).)

Background: Scales are composed of exfoliated keratinocytes and are frequently a secondary change, requiring the determination of the underlying cause. Consequently, making an appropriate differential diagnosis of scales and determining treatment necessity based on the results is crucial in dermatological practice., Case Description: We present a 7-year-old, neutered male Toy Poodle examined at Myogadani Animal Hospital with chief complaints of exercise intolerance, unexplained weight gain, and dermatological signs, including symmetrical alopecia of the trunk. The dog was diagnosed with hypothyroidism and administered levothyroxine sodium. A significant number of scales were observed, primarily on the trunk, after 1 month of administration. The scales disappeared after only reducing the dose of levothyroxine sodium for 3 months. Additionally, hair regrowth was observed as the scales disappeared. In this case, the scales were primarily observed in the alopecia area of the trunk, where hair growth was subsequently noted. Hence, levothyroxine sodium administration was thought to not only facilitate hair regrowth but also influence skin turnover, causing the appearance of scales., Conclusion: Clinicians who treat hypothyroidism need to be cognizant that symmetrical scales, primarily located on the trunk, do not co-occur with other skin lesions, such as erythema, and are not itchy, may appear within a month. The scales will naturally disappear in such cases when hair growth is complete; thus, no additional treatment is required., Competing Interests: The authors declare that there is no conflict of interest.

Background: Lung tumors in dogs, significantly primary paragangliomas, are rare and have not been reported. This report describes a dog with a lung tumor diagnosed as a primary paraganglioma., Case Description: A 12-year-old spayed French bulldog presented with a left-sided pulmonary mass. The dog was in good general condition and had no clinical symptoms. Computed tomography (CT) revealed a pulmonary mass near the bifurcation of the posterior lobe bronchus of the left lung. The mass showed a strong contrast enhancement effect that was subsequently attenuated. The dog underwent Surgery to remove the mass from the left lung. Abnormal hypertension was observed during surgery, and hypertensive crisis was suspected. Based on the histopathology and preoperative and postoperative urinary metanephrine and normetanephrine levels, the dog was diagnosed with primary paraganglioma of the lung. Although the CT scan showed findings suggestive of the development of a neuroendocrine tumor, it was difficult to suspect the development of a paraganglioma., Conclusion: The possibility of catecholamine-producing tumors should be considered when we encounter a lung tumor with no clinical symptoms and a neuroendocrine tumor-like contrast enhancement pattern on a CT scan., Competing Interests: The authors declare that there is no conflict of interest.

This case report discusses a 12-year-old West Highland White Terrier presented to the veterinary clinic with reverse sneezing and oral dysphagia. The endoscopic examination of the upper respiratory tract revealed a mass in the area of the right tonsil. Histologic examination confirmed the diagnosis of a malignant pilomatrixoma. The dog underwent surgical cytoreduction, followed by radiation therapy. After the treatment, the dog was free of clinical signs for 154 days and in good general condition. Due to recurrence of signs and the suspicion of clinically overt metastases in the mandibular lymph nodes, the dog was euthanized 261 days after the diagnosis. This is the first report of a malignant pilomatrixoma in the tonsillar area that has been treated with cytoreductive surgery and radiation therapy., Competing Interests: Die Autoren erklären hiermit, dass kein Interessenkonflikt besteht., (Thieme. All rights reserved.)

Three dogs were presented with a sudden onset of erythematous skin lesions, fever, and various extracutaneous signs, assigned to canine sterile neutrophilic dermatosis by clinical, laboratory, and histopathologic examination. This disease is very rare and comparable to Sweet syndrome in humans. According to the 4 forms of SS, the 1st and 3rd cases in this case report could be classified as classic/idiopathic SS. In this context, the respiratory signs may be a prodromal stage of SS preceding the skin lesions or possibly an infection of the respiratory tract was the trigger for SS. The 2nd case, on the other hand, shows a clear and the 3rd case a possible connection to pathergy as a consequence of a previous surgical treatment (case 2 a tibial plateau levelling osteotomy [TPLO] 20 days previously, case 3 a hemilaminectomy 90 days previously). This way, both may be assigned to the 4th form of SS. In all 3 cases, an adverse drug reaction as a trigger cannot be ruled out with certainty, as medication was used before the diagnosis was made in each case. Besides the most common extracutaneous signs such as fever and neutrophilia, immune-hemolytic anemia occurred in 2 of the patients. As therapeutic options, in addition to the established immunosuppressive drugs such as prednisolone, ciclosporin and azathioprine, oclacitinib is also included in the treatment regimen. In addition, administration of an antiplatelet agent (clopidogrel), which may be interpreted as a lesson learnt from the first of the presented cases, that died of pulmonary thromboembolism. In case 2, all drugs were tapered and eventually stopped over a period of 2.5 years and the patient has been without recurrence for 4 years. Case 3 is recurrence-free under therapy., Competing Interests: Die Autorinnen erklären, dass kein Interessenkonflikt besteht., (Thieme. All rights reserved.)

A 9-year-old castrated male German shepherd dog was presented because of a 2-day history of lethargy and anorexia. Abdominal distention and free peritoneal fluid were noted on physical examination, which prompted radiographs revealing a large, soft tissue and stippled gas opaque structure in the right cranial abdomen. Computed tomography was used to further describe the structure and assess for comorbidities in the dog. Both CT and abdominal radiographs supported a diagnosis of emphysematous splenic torsion. The enlarged spleen was surgically extracted, along with sections of necrotic omentum. Four days after the procedure, the dog developed a portal vein thrombus and secondary abdominal effusion, which ultimately led to the decision for humane euthanasia. Veterinarians should know the different imaging modalities used to diagnose splenic torsion and the possible postoperative complications following treatment., (Copyright and/or publishing rights held by the Canadian Veterinary Medical Association.)

A 4-year-old female Maltese dog was referred to our veterinary hospital with uveitis and conjunctivitis of the right eye. An ophthalmological evaluation revealed an intraocular mass that appeared to originate from the anterior uvea. Metastasis and regional invasion were not detected with CT examination. Enucleation of the right eye was recommended; however, the owner declined treatment. Six months later, the dog was re-presented with a right facial mass. At presentation, superficial lymph node enlargement was not appreciated, and no apparent alterations were noted on blood analysis or urinalysis. Computed tomography revealed an intraocular mass that invaded the surrounding tissues, including the frontal sinus. Presumed solitary ocular lymphoma with a large B-cell phenotype and Mott cell change was diagnosed via histopathological and immunohistochemical examination of a biopsy of the lesion. As the mass was too large for complete excision, neoadjuvant chemotherapy was administered. Complete remission was achieved using the L-COAP protocol and successful exenteration of the right eye. However, the dog was returned with enlargement of the right retropharyngeal lymph nodes. To the best of our knowledge, this is the first case report of presumed solitary ocular lymphoma with a large B-cell phenotype displaying Mott cell change in a dog. Key clinical message: This is the first reported case of a presumed solitary ocular lymphoma with a large B-cell phenotype and Mott cell change. Although systemic involvement was observed 6 mo after the initial visit, neoadjuvant chemotherapy and exenteration were effective., (Copyright and/or publishing rights held by the Canadian Veterinary Medical Association.)

A 3-year-old neutered male standard poodle mix dog was presented because of acute onset vomiting, increased respiratory effort, hyporexia, and progressive lethargy. On physical examination, intermittent tachycardia, tachypnea with mildly increased respiratory effort, and bilateral ventral muffled lung sounds were noted. Thoracic and abdominal computed tomography with contrast revealed a peritoneal pericardial diaphragmatic hernia (PPDH) with herniation of the liver and gallbladder, moderate abdominal lymphadenopathy, and mild pleural effusion. The dog underwent an emergency laparotomy and median sternotomy. Right ventral PPDH with a herniated, necrosed, and nonviable quadrate liver lobe and gallbladder; 2 suspected right ventricular (RV) aneurysms; and generalized mesenteric and portal lymphadenopathy were present. The affected liver lobe and gallbladder were removed; suspected aneurysms were managed by placing 2 purse-string sutures around the lesions and anchoring a pericardial flap over the aneurysms. The dog developed a ventricular arrhythmia postoperatively. Due to the arrhythmia and intraoperative findings of suspected aneurysms, echocardiography was performed and revealed focal RV systolic dysfunction, left ventricular systolic dysfunction, mild left ventricular dilation, and a hyperechoic area on the RV free wall, consistent with the purse string. An angiotensin-convertingenzyme (ACE) inhibitor was prescribed for left ventricular dilation. The dog was discharged 4 d postoperatively and was doing well 12 mo postoperatively. To our knowledge, this is the first report of a PPDH and suspected concurrent RV wall aneurysm in a dog successfully treated with a purse string and pericardial flap. Key clinical message: An RV aneurysm is extremely rare yet can be life-threatening in small animals. Early detection and treatment may minimize the risk of aneurysm rupture and sudden death., (Copyright and/or publishing rights held by the Canadian Veterinary Medical Association.)

A 13-year-old neutered male Lagotto Romagnolo dog had ultrasonographic and computed tomographic findings of pneumatosis intestinalis after presentation to the emergency department because of progressive diarrhea and hyporexia. Further investigations produced a diagnosis of multicentric lymphoma, and treatment with chemotherapy was commenced. Clinical remission of lymphoma was achieved and coincided with resolution of pneumatosis intestinalis on subsequent computed tomographic imaging. Key clinical message: Consider underlying pathology such as neoplasia in cases of pneumatosis intestinalis after excluding surgical emergent causes., (Copyright and/or publishing rights held by the Canadian Veterinary Medical Association.)

Parkinson's Disease (PD) is a clinically diagnosed disease that carries a reported misdiagnosis rate of 10-20%. Recent scientific discoveries have provided evidence of volatile organic compounds in sebum that are unique to patients with PD. The primary objective of this study was to determine if companion dogs could be trained to distinguish between sebum samples provided by PD-positive patients and PD-negative human controls. This was a randomized, handler-blind, controlled study. Twenty-three canines of varying breeds, ages, and environmental backgrounds were included. The study period encompassed 200 total working days from 2021 to 2022. Factors investigated included donor gender and levodopa drug affectivity, as well as canine breed, age, and duration of training time. The findings in this study were compiled from data collected during the final two years of a seven-year research program. For this two-year reporting period, when averaged as a group, the 23 dogs were 89% sensitive and 87% specific to olfactory distinction between PD-positive and PD-negative human donor samples. Ten of the twenty-three dogs averaged 90% or higher in both sensitivity and specificity. In 161 separate trials, a dog was presented with both novel PD-positive and PD-negative samples. For these novel exposures, the dogs collectively averaged 86% sensitivity and 89% specificity. PD medication was also investigated and was found to have no discernible impact on canine sensitivity or specificity results. Study findings support the application of companion dogs, trained with force-free, reward-based methodologies, for the detection of PD-positive and PD-negative samples under controlled conditions., (© 2024. The Author(s).)

Here we described a case of fatal canine visceral leishmaniasis (VL) in French Guiana, a non-endemic VL Amazonian area. The dog was a 2-year-old pug imported from Brazil to French Guiana. Initially seen for a pruriginous lesion on the muzzle which healed after treatment, the dog was in a deteriorated condition and had sublingual, foreleg and eye ulcers, one month later. A visceral leishmaniasis was suspected by the veterinarian. The dog was hospitalized awaiting results, which revealed the presence of L. infantum. However, the dog succumbed suddenly before the results were returned. Few imported and scarce autochthonous canine VL cases have been previously reported in French Guiana, raising the need for local epidemiological surveillance, considering the possibility of unusual transmission routes of the parasite., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Canine Visceral Leishmaniasis (CVL) is the most fatal form of Leishmania infection in dogs and is caused by L. infantum in the Americas. This parasite follows a zoonotic life cycle, raising concerns within domestic households, where dogs act as the primary reservoir of the parasite. Accurately detecting infected dogs is vital for effective epidemiological control in both canine and human populations. However, existing diagnostic methods in Brazil have limitations, particularly in detecting asymptomatic and oligosymptomatic dogs, leading to ineffective disease control. To address this challenge, we evaluated a novel recombinant antigen from L. infantum, the rLiNTPDase2. Previous studies have confirmed its high performance via ELISA, leading us to assess its suitability for a Lateral Flow Immunochromatographic Assay (LFIA), which is ideal for point-of-care testing. Standardization of the assay involved testing two nitrocellulose membranes (HF135 and HF120, Millipore), three blocking protocols, and five sample dilutions (1:10, 1:20, 1:40, 1:80, and 1:160). Following the chosen conditions (HF120 membrane, 1-minute blocking protocol, and 1:80 sample dilution), we validated our assay with a sample size of 78 dogs, comprising 32 negatives and 46 positives, including symptomatic (n=23), oligosymptomatic (n=17), and asymptomatic (n=6) cases. The results revealed a sensitivity of 86.9 %, specificity of 62.5 %, and accuracy of 76.9 %, which is consistent with ELISA performance for the same samples. Compared to DPP-LVC, our assay demonstrated promising results in detecting asymptomatic and oligosymptomatic cases. This study underscores the suitability of the rLiNTPDase2 antigen for the LFIA format, suggesting its potential as a novel point-of-care diagnostic test for CVL., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The urinary normetanephrine to creatinine ratio (uNMN/Cr) and urinary metanephrine to creatinine ratio (uMN/Cr) are commonly evaluated for the diagnosis of pheochromocytomas (PCC) in dogs. This study aimed to establish reference intervals for uNMN/Cr and uMN/Cr in 56 healthy dogs in Japan and to investigate the effect of urine collection methods on these measurements in 15 non-PCC dogs. The upper limits of reference intervals for uNMN/Cr and uMN/Cr were 124.4 nmol/mmol (90% confidence interval [CI] 107.7-137.0) and 121.1 nmol/mmol (90% CI 102.8-145.1), respectively. Both uNMN/Cr and uMN/Cr were significantly higher when urine was collected in the hospital compared to at home. Several factors, including the method of urine collection, should be considered when utilizing these reference intervals.

Hepatozoonosis, caused by the protozoan Hepatozoon canis, is a prevalent blood disease affecting owned and stray dogs and cats. The prevalence of these parasites among companion animals in Thailand remains poorly understood. Diagnosing the old-world form of the disease is challenging due to the wide range of nonspecific clinical signs and the reliance on finding low levels of Hepatozoon gamonts in blood smears for conventional diagnosis. PCR demonstrates high specificity and sensitivity but it requires sophisticated instrumentation. Therefore, we established recombinase polymerase amplification (RPA) coupled with Cas12a for H. canis detection based on 18S rRNA. Our findings showed that RPA-Cas12a using gRNA&#95;H was highly specific to H. canis, without yielding positives for other pathogen species including Babesia species. Even in cases of co-infection, RPA-Cas12a only detected positives in samples containing H. canis. This approach detected minimal amounts of H. canis18S rRNA-harboring plasmid at 10 copies per reaction, whereas plasmid-spiked canine blood enabled detection at a minimal amount of 100 copies per reaction. The performance of RPA-Cas12a was validated by comparing it with quantitative PCR-high resolution melting analysis (qPCR-HRM) and sequencing based on 35 canine blood samples. RPA-Cas12a demonstrated precision and accuracy values of 94 % and 90 %, respectively comparable to qPCR-HRM. Overall, these results indicate that RPA-Cas12a serves as a promising tool for H. canis detection as indicated by comparable performance to qPCR-HRM and is suitable for implementation in small animal hospitals or clinics due to its minimal resource requirements, thereby contributing to effective diagnosis and treatment for infected dogs., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Kittisak Buddhachat reports financial support was provided by National Research Council of Thailand (NRCT), Thailand. Korakot Nganvongpanit reports a relationship with Excellence Center in Veterinary Bioscience, Chiang Mai University, Chiang Mai, Thailand that includes: board membership and employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 Elsevier B.V. All rights reserved.)

An 11-year-old, female, spayed, soft-coated Wheaten terrier presented for acute onset of neurological signs. On presentation, neurological examination showed right thoracic and pelvic limb proprioceptive deficits, absent right menace reflex, and weak right nasal septum response. A left thalamocortical lesion was localized. On thoracic auscultation, an arrhythmia was noted, and electrocardiography showed frequent ventricular premature complexes and rare runs of ventricular tachycardia. Echocardiography identified an interventricular septal mass extending into the lumen of the left ventricle. Thalamocortical metastasis secondary to the cardiac mass was suspected to be the cause of the patient's neurological signs. Humane euthanasia was elected by the owner due to the patients clinical status and poor prognosis. A postmortem examination diagnosed hemangiosarcoma of the interventricular septum, the right ventricular free wall, and left ventricular free wall. The left ventricle adjacent to the paraconal groove showed myocardial necrosis and inflammation. Metastases to the brain and secondary intracranial hemorrhage were found which were suspected to be the cause of the antemortem neurological signs. Concurrent pulmonary and hepatic metastases were noted. This report describes a rare presentation of an intracardiac hemangiosarcoma of the interventricular septum, right ventricle, and left ventricle in a patient presenting with neurological signs., Competing Interests: Conflict of Interest Statement The authors do not have any conflicts of interest to disclose., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Canine leishmaniosis (CanL), caused by Leishmania sp., presents a wide array of symptoms; renal dysfunction is frequently observed in these dogs and is associated with a poor prognosis and increased mortality. The traditional biomarkers namely urea and creatinine can detect renal damage but only in advanced stages of the disease. However, it has been shown that the symmetric dimethylarginine assay (SDMA) or the protein/creatinine ratio (UPC) and are early biomarkers of renal dysfunction. Their elevation occurs earlier than that of creatinine, but other novel biomarkers such as neutrophil gelatinase-associated lipocalin (NGAL) are currently under investigation. Our objective was to determine whether the urine NGAL-creatinine ratio (uNGAL/c) can provide very early diagnosis of kidney disease in CanL. In total, 68 dogs were included in the study: 15 healthy dogs and 53 dogs with CanL who were classified according to International Renal Interest Society (IRIS) classification: IRIS 1 (N= 34), IRIS 2 (N= 9) and IRIS 3/4 (N= 10). IRIS 1 was subdivided according to proteinuria in IRIS 1 NP (13 dogs with UPC < 0.2), IRIS 1 BL (8 dogs with UPC = 0.2-0.5) and IRIS 1 P (13 dogs with UPC > 0.5). Blood samples were collected for complete hematological and biochemistry analysis including plasma NGAL. Urinalysis included specific gravity, UPC, CysC and NGAL expressed as a ratio with creatinine. The mean concentrations of pCysC and SDMA in CanL, show a statistically significant increase from IRIS 1 NP , not being statistically significant for pCysC in the IRIS 1 BL group. The UPC show a statistically significant increase from IRIS 1 NP. In all groups with CanL for uCysC/c and uNGAL/c was observed a statistically significant increase. The uNGAL/c in the group proteinuric animals, presents a positive correlation with all renal biomarkers studied. In the group of non-proteinuric animals, the uNGAL/c presents a positive correlation with SDMA and UPC. The uNGAL/c can be considered a reliable indicator of renal disease in dogs diagnosed with CanL who are non-azotemic and non-proteinuric., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

This report describes a suspected case of paraneoplastic syndrome of inappropriate antidiuretic hormone (SIADH) in a 5-year-old male entire Australian Kelpie associated with a gastric adenocarcinoma. The dog had a history of chronic vomiting and presented with hyponatraemia, hypokalaemia, hypochloraemia and increased urinary sodium. Computed tomography identified the presence of a gastric tumour and histopathology confirmed a pyloric gastric adenocarcinoma. Removal of the tumour via a Billroth I procedure resulted in rapid resolution of electrolyte derangements. The dog was still alive six months postoperatively with no recurrence of either SIADH or tumour at the time of publication., (© 2024 Australian Veterinary Association.)

A 4-month-old female Shar-pei dog was admitted with apathy, anorexia, and vomiting. The patient had an appropriate vaccination protocol, with the last vaccine administered 2.5 weeks prior to the onset of clinical signs. Physical examination revealed tachycardia, fever and swelling of the tibiotarsal joints. Several diagnostic tests including complete blood cell count, biochemistry profile, urinalysis, thoracic radiographs, hind limbs radiographs, abdominal ultrasound, and infectious diseases tests, were conducted to determine the underlying cause. Shar-Pei Auto-inflammatory Disease (SPAID) was diagnosed. Additionally, the patient developed skin necrosis in the inner aspect of the tibiotarsal joints as a complication. A skin biopsy revealed cutaneous vasculopathy causing degeneration, abrupt ulceration, and ischemic necrosis with intense neutrophilic inflammation of the dermis and subcutis. Moreover, a hospital-acquired infection was identified by skin culture. Debridement of the necrotic skin was performed, and due to its' severity and extent, the wound was closed by secondary intention. A diagnostic protocol and the therapeutic dose of acetylsalicylic acid, which led to clinical improvement, are recommended here. The patient has continued to present episodic manifestations of SPAID mainly fever and swelling of the tibiotarsal joints, but there has been no recurrence of necrosis or other cutaneous lesion in the last two years., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Cystic echinococcosis, resulting from infection with Echinococcus granulosus, poses a significant challenge as a neglected tropical disease owing to the lack of any known effective treatment. Primarily affecting under-resourced, remote, and conflict-ridden regions, the disease is compounded by the limitations of current detection techniques, such as microscopy, physical imaging, ELISA, and qPCR, which are unsuitable for application in these areas. The emergence of CRISPR/Cas12a as a promising tool for nucleic acid detection, characterized by its unparalleled specificity, heightened sensitivity, and rapid detection time, offers a potential solution. In this study, we present a one-pot CRISPR/Cas12a detection method for E. granulosus (genotype G1, sheep strain) integrating recombinase polymerase amplification (RPA) with suboptimal protospacer adjacent motif (PAM) and structured CRISPR RNA (crRNA) to enhance reaction efficiency. The evaluation of the assay's performance using hydatid cyst spiked dog feces and the examination of 62 dog fecal samples collected from various regions of Western China demonstrate its efficacy. The assay permits visual observation of test results about 15 minutes under blue light and displays superior portability and reaction speed relative to qPCR, achieving a sensitivity level of 10 copies of standard plasmids of the target gene. Analytic specificity was verified against four tapeworm species (E. multilocularis, H. taeniaeformis, M. benedeni, and D. caninum) and two other helminths (T. canis and F. hepatica), with negative results also noted for Mesocestoides sp. This study presents a rapid, sensitive, and time-efficient DNA detection method for E. granulosus of hydatid cyst spiked and clinical dog feces, potential serving as an alternative tool for field detection. This novel assay is primarily used to diagnose the definitive host of E. granulosus. Further validation using a larger set of clinical fecal samples is warranted, along with additional exploration of more effective approaches for nucleic acid release., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

SARS-CoV-2 emerged from an animal source and was then transmitted to humans, causing the COVID-19 pandemic. Since a wide range of animals are susceptible to SARS-CoV-2 infection, the zoonotic potential of SARS-CoV-2 increases with every new animal infected. The molecular gold standard assay for SARS-CoV-2 detection is real-time RT-PCR, where the Ct obtained is proportional to the amount of nucleic acid and can be a semi-quantitative measure of the viral load. However, since the use of real-time RT-PCR assays in animal samples is low due to the high costs, the use of validated nested PCR assays will help to monitor large-scale animal samplings, by reducing the costs of detection. In the present study, 140 samples from dogs and cats (15 SARS-CoV-2-positive samples with Ct values from 27 to 33, and 125 negative samples), previously analyzed by real-time RT-PCR, were analyzed by nested PCR. To increase the number of positive samples to determine the sensitivity of the assay, 40 human samples obtained during COVID-19 diagnosis in 2020 were included. The specificity of the primers was analyzed against samples positive to canine coronavirus (CCV) and feline infectious peritonitis virus (FIPV). To calculate the limit of detection (LoD) of the nested PCR, the viral load was estimated extrapolating the Ct value obtained by real-time RT-PCR. The Ct values obtained were considered as semi-quantitative and were able to distinguish between high, moderate and low viral loads. The Kappa value or "agreement" between assays and reliability of the nested PCR were also determined. Eleven of the animal samples analyzed by nested PCR targeting the N gene were detected as positive, while 129 were detected as negative to the virus, with Ct values ranging between17 and 31.5. All the samples from humans analyzed by nested PCR were positive. These results indicate that the assay has a sensitivity of near 95 % and a specificity of 100 %. No unspecific reactions analyzed by nested PCR were observed with the samples positive to CCV and FIPV. The samples detected as positive to SARS-CoV-2 by nested PCR were those that presented a Ct between17 and 31.5. The LoD of the nested PCR was estimated close to 50 copies/µL of viral load, corresponding with a Ct of 31.5. The Kappa value between assays was excellent (k = 0.829). The results obtained demonstrate that nested PCR is useful to detect SARS-CoV-2 low viral loads at a lower cost than with real-time RT-PCR., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)