Evidence of exposure to SARS-CoV-2 in cats and dogs from households in Italy.

SARS-CoV-2 emerged from animals and is now easily transmitted between people. Sporadic detection of natural cases in animals alongside successful experimental infections of pets, such as cats, ferrets and dogs, raises questions about the susceptibility of animals under natural conditions of pet ownership. Here, we report a large-scale study to assess SARS-CoV-2 infection in 919 companion animals living in northern Italy, sampled at a time of frequent human infection. No animals tested PCR positive. However, 3.3% of dogs and 5.8% of cats had measurable SARS-CoV-2 neutralizing antibody titers, with dogs from COVID-19 positive households being significantly more likely to test positive than those from COVID-19 negative households. Understanding risk factors associated with this and their potential to infect other species requires urgent investigation.

Evidence of exposure to SARS-CoV-2 in cats and dogs from households in Italy.

SARS-CoV-2 originated in animals and is now easily transmitted between people. Sporadic detection of natural cases in animals alongside successful experimental infections of pets, such as cats, ferrets and dogs, raises questions about the susceptibility of animals under natural conditions of pet ownership. Here we report a large-scale study to assess SARS-CoV-2 infection in 817 companion animals living in northern Italy, sampled at a time of frequent human infection. No animals tested PCR positive. However, 3.4% of dogs and 3.9% of cats had measurable SARS-CoV-2 neutralizing antibody titers, with dogs from COVID-19 positive households being significantly more likely to test positive than those from COVID-19 negative households. Understanding risk factors associated with this and their potential to infect other species requires urgent investigation. ONE SENTENCE SUMMARY: SARS-CoV-2 antibodies in pets from Italy.

Cytomorphological description and intra-observer agreement in whole slide imaging for canine lymphoma.

Whole slide imaging (WSI) uses robotic microscopes for computerising entire slides into digital images. The aim of this study was to assess the agreement between WSI and optical microscopy for evaluating canine lymphoma cytological samples. Forty-four slides were computerised using a WSI scanner and the digital and glass slides were examined by three observers with different levels of expertise. Morphology and grade of lymphoma were scored on the basis of the updated Kiel classification and intra-observer agreement was assessed. The accuracy of determining the grade of lymphoma with digital and glass slides based on the results of flow cytometry (FC) was established. The overall intra-observer agreement for cytomorphological features was fair to moderate (κ=0.34-0.52) for the three observers and moderate (κ=0.44-0.53) for the evaluation of grade of malignancy. The diagnostic agreement between FC and digital slides was slight (κ=0.16) for the inexperienced observer, fair (κ=0.32) for the mildly experienced observer and moderate (κ=0.50) for the very experienced observer. The diagnostic agreement between FC and glass slides was fair (κ=0.37) for the inexperienced observer, substantial (κ=0.63) for the mildly experienced observer and moderate (κ=0.50) for the very experienced observer. These findings underline the importance of observer experience in determining the grade of malignancy, especially if digital slides are used. The study also identifies some technical limitations of the WSI scanner used in this study, mainly linked to image quality, which might affect the morphological evaluation of neoplastic cells.

Effects of Benazepril on Survival of Dogs with Chronic Kidney Disease: A Multicenter, Randomized, Blinded, Placebo-Controlled Clinical Trial.

BACKGROUND: Chronic kidney disease (CKD) is an important cause of morbidity and mortality in dogs. OBJECTIVE: To evaluate the efficacy in prolonging survival and safety of benazepril administration to dogs with CKD. ANIMALS: Forty-nine client-owned dogs with CKD. METHODS: Dogs were randomized to benazepril (0.25 to <0.5 mg/kg) or placebo once daily for up to 2 years in a prospective, multicenter, blinded clinical trial. The primary endpoint variable was the renal survival time, defined as the time from inclusion in the study to the treatment failure endpoint of death or euthanasia or need for administration of parenteral fluids related to renal failure. RESULTS: No benefit of benazepril versus placebo was detected for renal survival time in all dogs; median (95% confidence interval (CI)) survival times were 305 (53-575) days in the benazepril group and 287 (152-not available) in the placebo group (P = .53). Renal survival times were not significantly longer with benazepril compared to placebo for subgroups: hazard ratios (95% CI) were 0.50 (0.21-1.22) with P = .12 for initial urine protein-to-creatinine ratio (UPC) >0.5, and 0.38 (0.12-1.19) with P = .080 for initial UPC >0.5 plus plasma creatinine ≤440 µmol/L. Proteinuria, assessed from the UPC, was significantly (P = .0032) lower after treatment with benazepril compared to placebo. There were no significant differences between groups for clinical signs or frequencies of adverse events. CONCLUSIONS AND CLINICAL RELEVANCE: Benazepril significantly reduced proteinuria in dogs with CKD. Insufficient numbers of dogs were recruited to allow conclusions on survival time.

An open-label phase 1 dose-escalation clinical trial of a single intravenous administration of gemcitabine in dogs with advanced solid tumors.

BACKGROUND: A broad range of gemcitabine dosages have been used in dogs. HYPOTHESIS/OBJECTIVES: To determine maximally tolerated dose (MTD), dose-limiting toxicity (DLT), and preliminary antitumor activity of intravenous administration of gemcitabine in dogs with advanced solid tumors. ANIMALS: Twenty-two client-owned dogs. METHODS: Dogs with advanced cancer were prospectively enrolled in an open-label Phase 1 study of gemcitabine. Gemcitabine was administered as a 30-minute intravenous bolus starting at 800 mg/m(2), using escalation of 50 mg/m(2) increments with 3 dogs per dose level. MTD was established based on the number of dogs experiencing DLT assessed after 1 cycle. Treatment continued until disease progression or unacceptable toxicosis. Additional dogs were enrolled at MTD to better characterize tolerability, and to assess the extent and duration of gemcitabine excretion. RESULTS: Twenty-two dogs were treated at 4 dose levels, ranging from 800 to 950 mg/m(2). Neutropenia was identified as DLT. MTD was 900 mg/m(2). DLT consisting of grade 4 febrile neutropenia was observed at 950 mg/m(2) in 2 dogs. There were no nonhematologic DLTs. Twenty dogs received multiple doses, and none had evidence of severe toxicosis from any of their subsequent treatments. At 900 mg/m(2), 2 complete and 5 partial responses were observed in dogs with measurable tumors. The amount of gemcitabine excreted in urine decreased over time, and was undetectable after the first 24 hours. CONCLUSIONS AND CLINICAL IMPORTANCE: The recommended dose of gemcitabine for future Phase 2 studies is weekly 900 mg/m(2). In chemotherapy-naïve dogs with advanced solid tumor this dose level merits further evaluation.

Diagnostic value of cytological analysis of tumours and tumour-like lesions of the oral cavity in dogs and cats: a prospective study on 114 cases.

Neoplastic or non-neoplastic masses are common findings in the oral cavity of cats and dogs. The aim of this prospective study was to compare the results of cytological examinations of lesions of the oral cavity following fine-needle aspiration (FNA), fine-needle insertion (FNI), and impression smear (IS) with histopathological results being considered as the diagnostic gold standard. In total, 85 dogs and 29 cats were included in the study. Cases were included when histology and cytology (FNA, FNI, and/or IS) were available from the same lesion; κ-agreement and accuracy between cytological and histopathological results were calculated. Eighteen cytological specimens were excluded, with a retrieval rate of 84.2%. Of the 96 samples analysed, FNA, FNI, and IS were available from 80, 76, and 73 animals, respectively. Overall, 60/67 (89.6%) and 21/29 (72.4%) lesions were neoplastic in dogs and cats, respectively, with the remaining being non-neoplastic. For all lesions, κ-values obtained by FNA, FNI, and IS were in dogs 0.83 (95% confidence interval [CI]: 0.77-0.90), 0.87 (95% CI: 0.81-0.93) and 0.75 (95% CI: 0.67-0.84), respectively, and in cats 0.92 (95% CI: 0.87-0.96), 0.92 (95% CI: 0.88-0.97) and 0.86 (95% CI: 0.79-0.92), respectively. The diagnostic accuracies of FNA, FNI, and IS in dogs with neoplasia were 98.2%, 98.1%, and 91.8%, respectively, and in cats with neoplasia were 95.6%, 95.6% and 95.8%, respectively. In conclusion, the high agreement with histopathology suggests that cytological examinations by FNI, FNA, and IS are all appropriate methods to correctly diagnose lesions of the oral cavity in dogs and cats.

Cytological and histological correlation in diagnosing feline and canine mediastinal masses.

OBJECTIVES: The aim of this study was to evaluate the agreement between cytological and histological diagnosis of canine and feline mediastinal masses to assess the utility of cytological examination in accurately diagnosing and classifying mediastinal lesions. METHODS: A retrospective review of 58 cases of mediastinal masses from 21 dogs and 37 cats were performed. Histopathology was used as the diagnostic reference standard. The agreement between cytological and histological diagnosis was calculated. RESULTS: The complete agreement between cytological and histological classification ranged from substantial (k = 0 · 72, CI: 0 · 64 to 0 · 80) to almost perfect (k = 0 · 89, CI: 0 · 82 to 0 · 96) depending on how the cytological diagnoses classified as suspicious were used for statistical calculations. CLINICAL SIGNIFICANCE: Cytological examination of canine and feline mediastinal masses is a relatively easy, low-cost procedure, with good agreement with final histological diagnoses.

Torsion of quadrate and right middle liver lobes and gallbladder in a German Shepherd dog.

BACKGROUND: An 11-year-old female German Shepherd was referred for assessment and management of a suspected liver tumour. Abdominal ultrasonography revealed abundant abdominal effusion and a right liver mass in continuity with a cystic structure. The dog had undergone splenectomy for a low-grade lymphoma 1 year prior. METHODS: Serum biochemical analysis revealed only a mild increase in alanine aminotransferase. The analysis of the free abdominal fluid showed a modified transudate. Ultrasound confirmed the presence of a hypoechoic lesion involving the right liver lobes, in addition to an over-distended gallbladder. Computed tomography examination demonstrated a massively dilated gallbladder and lower attenuation of the adjacent liver lobes, with no enhancement on quadrate and right middle liver lobes. The fine needle aspirate was compatible with a mixed subacute to chronic inflammation. Based on the investigational findings, an exploratory coeliotomy was performed. TREATMENT AND OUTCOME: The middle right and quadrate liver lobes and the gallbladder were rotated on their axes. The torsed lobes and the cystic duct were removed using a TA stapler without derotating the organs. Histopathological results were suggestive of infarction secondary to torsion of the lobes and gallbladder. The dog recovered from surgery without complications, and 20 months later was in good overall condition. CONCLUSION: Liver lobe torsion is an uncommon condition that rarely affects the right lobes. Gallbladder torsion is even more uncommon. To our knowledge, this is the first recorded case of quadrate and right middle liver lobe and gallbladder torsion.

Cytosine arabinoside in addition to VCAA-based protocols for the treatment of canine lymphoma with bone marrow involvement: does it make the difference?

Cytosine arabinoside (ara-C) is a component of many protocols for the treatment of acute leukaemia and non-Hodgkin lymphomas in humans. The aim of the study was to prospectively evaluate the efficacy of ara-C in a myeloablative regimen in a cohort of canine lymphomas with bone marrow involvement. Seventeen dogs were enrolled. Eight were treated with a VCAA-based protocol (Group 1) and nine with the same regimen added with ara-C (Group 2). Ara-C was administered on a 5-day schedule as an i.v. continuous infusion at the dose of 150 mg m(-2) per day for five consecutive days. During treatment complete remission (CR) was achieved in two dogs in Group 1 and in eight dogs in Group 2. CR rate was significantly higher in Group 2 (P < 0.01). Median survival was 72.5 days (range 6-174) in Group 1 and 243 days (range 73-635) in Group 2. Survival was significantly longer in Group 2 (P < 0.001). Both protocols were well tolerated, with a low incidence of adverse events. Ara-C added to a VCAA-based protocol appears to be safe and beneficial in dogs with stage V lymphoma. Incorporation of the nucleoside analogue might be crucial for the development of future therapeutic strategies in dogs.

Advanced glycation end products and sorbitol in blood from differently compensated diabetic dogs.

Canine diabetes mellitus (DM) is a common metabolic disorder with long term complications, most of which are caused by glycosylation of structural proteins, decreases in antioxidant concentrations, altered osmotic balance and hypoxia due to impaired oxygen transport. Previous studies have demonstrated that under hyperglycemic conditions canine erythrocytes undergo swelling, probably due to activation of the polyol pathway. The present work aimed to assess the plasma concentration of advanced glycation end (AGE) products, stable Amadori-products generated by non-enzymatic glycosylation of proteins and the intracellular concentration of sorbitol, produced by the activation of polyol pathway in 34 blood samples from diabetic dogs and in 14 controls. AGE products were significantly higher (p<0.01) in plasma from diabetic dogs compared with control animals. The sorbitol concentration in erythrocytes was also significantly higher in diabetic dogs and, in particular, in poorly compensated animals and in dogs with ketonuria. In five cases that were analysed before and after clinical improvement, sorbitol concentration was found to correlate with improvement. These results suggest that non-specific glycosylation is increased and that the polyol pathway is activated in diabetic dogs in a manner that is proportionate to the severity of disease. Moreover, the concentration of AGE products and sorbitol may be useful for monitoring the onset of diabetic complications and assessing the most appropriate therapeutic approaches for management of canine DM.

Unusual dermatological toxicity of hydroxyurea in two dogs with spontaneously occurring tumours.

Hydroxyurea is a chemotherapeutic agent used to treat hypereosinophilic syndrome, mast cell tumours and many myeloproliferative disorders. It is usually well tolerated; however, reported side effects include myelosuppression and gastrointestinal complications as well as cutaneous toxicity, in very rare cases. We report the unusual appearance of onychomadesis involving several claws on all feet in two canine patients receiving long-term hydroxyurea treatment. The healing of nail lesions required cessation of treatment in one dog and dose reduction in the second animal, supporting a strict relationship between onychopathy and administration of hydroxyurea. The aim of this case report is to increase clinical awareness of dermatological toxicity associated with long-term administration of hydroxyurea.

Nasal melanosis in three dogs.

Cytologically and histologically confirmed nasal melanosis was detected by rostrocaudal rhinoscopic evaluation of three dogs with unilateral nasal discharge caused by a chronic and severe odontopathic rhinitis. The extraction of affected teeth and prolonged antibiotic therapy led to a complete resolution of nasal disease. The nasal melanosis could be considered a partial metaplastic transformation of mucosal respiratory cells with accumulation of intracytoplasmic melanin.

Diagnosis of canine nasal aspergillosis by cytological examination: a comparison of four different collection techniques.

OBJECTIVES: To compare the efficacy and diagnostic value of four different sample collection techniques for cytological identification of nasal aspergillosis-penicilliosis in dogs. METHODS: Fifteen dogs with a history of persistent nasal discharge and clinical and radiographic findings suggestive of aspergillosis were evaluated using four different cytological sampling techniques. These were a direct smear from the nasal discharge, blind swab collection under general anaesthesia, brushing from suspect lesions under direct endoscopic visualisation and a squash technique of mucosal biopsies from suspect lesions obtained under direct endoscopic visualisation. RESULTS: Direct smear collection and blind swab collection detected fungal hyphae in 13.3 and 20 per cent of examined cases, respectively; brush samples detected fungal hyphae in 93.3 per cent and fungal spores in the 45 per cent of examined cases and squash samples detected fungal hyphae in 100 per cent and fungal spores in 36 per cent of examined cases. CLINICAL SIGNIFICANCE: This study confirmed the high accuracy of cytology samples in the diagnosis of nasal aspergillosis-penicilliosis when collected under direct endoscopic visualisation and showed the poor value of samples that were collected by blind swabs or prepared from samples of nasal discharge.

Correlation between fine-needle aspiration cytology and histopathology in the evaluation of cutaneous and subcutaneous masses from dogs and cats.

BACKGROUND: Fine-needle aspiration cytology (FNAC) is commonly used as a diagnostic procedure to evaluate superficial and deep masses in animals. However, few studies have addressed the accuracy of FNAC in the evaluation of cutaneous and subcutaneous masses in a clinical setting. OBJECTIVE: The purpose of this study was to compare the accuracy of FNAC as compared with histopathology in the diagnosis of cutaneous and subcutaneous masses from dogs and cats. METHODS: Cytologic and histopathologic specimens obtained between 1999 and 2003 from 292 palpable cutaneous and subcutaneous masses obtained from 242 dogs and 50 cats were retrospectively evaluated. Cytologic samples were obtained by FNA and histopathologic samples were collected by surgical biopsy or at necropsy. Concordance was determined and the accuracy of FNAC for the diagnosis of neoplasia was determined using histopathology as the gold standard. RESULTS: Of 292 specimens, 49 (from 44 dogs and 5 cats) were excluded due to poor cellularity of the cytologic specimen (retrieval rate 83.2%, n = 243). A cytologic diagnosis of neoplasia was obtained in 176 cases (175 true positives and 1 false positive compared with histopathology). Sixty-seven cytology samples were classified as non-neoplastic (46 true negatives, 21 false negatives compared with histopathology). Overall, the cytologic diagnosis was in agreement with the histopathologic diagnosis in 90.9% (221/243) of cases. For diagnosing neoplasia, cytology had a sensitivity of 89.3%, a specificity of 97.9%, a positive predictive value of 99.4%, and a negative predictive value of 68.7%. CONCLUSIONS: The results of this study confirmed FNAC as a reliable and useful diagnostic procedure for the evaluation of palpable cutaneous and subcutaneous lesions in small animal practice.

Preliminary studies of serum acute-phase protein concentrations in hematologic and neoplastic diseases of the dog.

Serum concentrations of acute-phase proteins (APPs): haptoglobin (Hp), ceruloplasmin (Cp), serum amyloid A (SAA), and C-reactive protein (CRP) were determined in healthy dogs (n = 15) and dogs with different diseases grouped as acute inflammation (I, n = 12), hematologic neoplasias (HT, including leukemia and lymphoma, n = 16), nonhematologic neoplasias (NHT, including epithelial, mesenchymal, and mixed, n = 20), and autoimmune hemolytic anemia (AIHA, n = 8). SAA and CRP were analyzed using commercially available enzyme-linked immunosorbent assay (ELISA) kits, and Hp and Cp were measured using colorimetric methods, all previously validated for use in dogs. Increased concentrations of all APPs were observed in all groups of diseased dogs, but statistical significance only was observed with Hp (I, P < .001; HT, P < .05), Cp (I, P < .05; AIHA, P < .01), and CRP (I, P < .001; HT, P < .001; AIHA, CRP P < .05). High variability in individual APPs within each group of diseases was found with no significant differences between leukemia and lymphoma as well as among different types of neoplasia. The AIHA group had smaller increases in Hp, SAA, and CRP but higher concentrations of Cp. When follow-up of individual cases was possible, a decrease in APPs generally was found in cases with favorable outcome. The results of this study suggest that neoplasia and hematologic diseases such as AIHA should be considered as possible causes of mild increases in APPs in dogs. Measurement of APPs may be helpful to assess clinical evolution and monitor treatment of these processes.

Cytologic features of testicular tumours in dog.

In this paper, we report on our experience of cytology of fine needle biopsies performed on 92 dogs with testicular tumours during the period from 1998 to 2002. Cytological diagnosis was consistent with seminoma in 20 cases, sertolioma in 16 cases, Leydig cell tumours in 50 cases and mastocytoma in one case. Five cases could not be diagnosed by cytology. Cytological observations were confirmed after surgery by histopathological examination in 87 cases. Cytology provided a sensitivity of 95% for seminoma, 88% for sertolioma and 96% for Leydig cell tumours. The specificity was 100% for all three tumour types. In our experience cytology of fine needle aspirations of testicular tumours is a very reliable technique.

Clinical, cytological and molecular evidence of Mesocestoides sp. infection in a dog from Italy.

A 12-year-old, 13 kg, mixed-breed male dog was referred for anorexia and depression. The dog showed discomfort on abdominal palpation. Abdominal ultrasound examination revealed multiple, small, round anechoic cystic structures. Cystic fluid obtained with fine needle aspiration contained several 2-4 mm white motile flecks. Microscopic examination of the fluid revealed numerous irregularly shaped organisms measuring several hundred microns to 3 mm, the morphology of which was suggestive of intact and fragmented acephalic metacestodes of the genus Mesocestoides sp. Molecular analysis confirmed that the peritoneal infection was caused by Mesocestoides sp.

Cytological features of canine ovarian tumours: a retrospective study of 19 cases.

The cytological features of 19 histologically confirmed canine ovarian tumours were retrospectively examined. Seven cases were cytologically classified as papillary adenocarcinoma, eight cases as granulosa cell tumours, two cases as mature ovarian teratomas, one case as a dysgerminoma and one case as a mixed granulosa cell tumour/dysgerminoma. On cytology, papillary adenocarcinoma was characterised by a papillary glandular pattern and tight cohesiveness. Granulosa cell tumours showed monolayered clusters of loosely cohesive granulosa cells. Call-Exner-like bodies were found in five of seven cases. Granulosa cells appeared to be heterogeneous and usually contained several intracytoplasmic vacuoles. Teratoma was characterised cytologically by keratin debris (two cases) and a mixture of epithelial cells with sebaceous, basaloid, columnar/palisading or ciliated appearance (one case). The dysgerminoma contained severely atypical round cells admixed with small lymphocytes. The mixed dysgerminoma/granulosa cell tumour had a mixture of germinal and granulosa cells. Cytological diagnosis was in agreement with histopathology in 18 of 19 (94.7 per cent) cases.

Percutaneous fine-needle biopsy of deep thoracic and abdominal masses in dogs and cats.

Percutaneous fine-needle biopsy was used to investigate thoracic and abdominal masses in the dog and cat. One hundred and thirty-two cases were included in the study; 20 cases were excluded from the comparative study due to poor cellularity or blood contamination (retrieval rate 86.8 per cent). One hundred samples (56 dogs and 44 cats) were classified by cytology as neoplastic. All the cytological diagnoses of neoplasia were confirmed by histological samples obtained either by non-surgical methods, at surgery or during postmortem examination. No false positive diagnoses of neoplasia were made. Thirty-two samples were cytologically classified as 'negative for neoplasia'. Subsequent histological examination revealed 18 true negative and 14 false negative results. The procedure had an overall 89.4 per cent (118 cases out of 132) agreement between the diagnosis of inflammatory disease versus neoplasia, with a sensitivity of 87.8 per cent, a specificity of 100 per cent, a predictive value of a positive test of 100 per cent and a predictive value of a negative test of 56.3 per cent.

Clinicopathological findings in five cats with paw calcification.

This retrospective study describes the clinicopathological findings in five cats with soft tissue mineralisation of interdigital spaces and footpads. Paw disease was the reason for veterinary consultation in three out of five cats. All cats had laboratory findings suggestive of renal failure and high solubility product [calciumxphosphorus]. In all cases, cytological examination of paw lesions was suggestive of calcinosis. The results of our study agree with two previous case reports of paw calcification in the cat, suggesting a metastatic pathogenesis and a correlation between paw mineralisation and renal failure.