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Canine bufavirus (CBuV) is a protoparvovirus, genetically related to human and non-human primate bufaviruses and distantly related to canine parvovirus type 2 (CPV-2). CBuV was initially identified from young dogs with respiratory signs but subsequent studies revealed that this virus is also a common component of the canine enteric virome. In this survey, by assessing archival and recent collections of dogs faecal samples, CBuV DNA was detected with a higher prevalence rate (8.8%) in animals with enteritis than in control animals (5.0%), although this difference was not statistically significant. The rate of co-infections with other enteric viruses in diarrhoeic dogs was high (84.6%), mostly in association with canine parvovirus CPV-2 (90.1%). The complete ORF2 gene was determined in five samples, and the nearly full-length genome was reconstructed for three strains, 62/2017/ITA, 9AS/2005/ITA and 35/2018/ITA. Upon sequence comparison, the viruses appeared highly conserved in the NS1 (97.2%-97.9% nt and 97.5%-98.1% aa identities). In the complete VP2 coding region, three strains were similar to the prototype viruses (99.7-99.8 nt and 99.6%-99.8% aa) whilst strains 9AS/2005/ITA and 35/2016/ITA were distantly related (87.6%-89.3% nt and 93.9%-95.1% aa identities). Interestingly, genetic diversification occurred downstream conserved regions such as the VP1/VP2 splicing signals and/or the G-rich motif in the N terminus of the VP2, suggesting a potential recombination nature. Upon phylogenetic analysis, the two divergent CBuV strains formed a distinct cluster/genotype.

Diagnostic imaging plays a fundamental role in the diagnosis of pulmonary diseases. Radiography, ultrasound, computed tomography, and endoscopy are important tools for achieving a diagnosis. The choice of diagnostic procedure varies according to the patient, the suspected diagnosis and the risk/benefit ratio. Culture, cytology and histology are nearly always necessary to obtain a definitive diagnosis. Several biopsy sampling techniques are described. Surgical biopsies are the gold standard for the diagnosis of bronchiolitis or interstitial lung diseases but often not performed due to the high risk. In humans, the introduction of transbronchial cryobiopsies has led to excellent results in the study of interstitial lung diseases.

The goal of this study was to report the first case of antral stenosis in a cat. We have described a clinical case of a 9 months old domestic shorthaired cat referred for persistent and refractory vomiting, anorexia and loss of weight. Ultrasound (US) examination detected a mild gastric distension, mild wall thickening (5mm), severe serosal irregularities and a constriction between the fundus and body. Computed tomography (CT) was then performed: a severe gastric stenosis between the body and antrum with minimum contrast transit was detected and confirmed by flexible endoscopic examination. The antral localization of the stenosis was revealed after a cranial celiotomy and Y-U plastic was necessary to correct the defect. The patient completely recovered after surgery with significant body weight gain and remained in remission throughout a 2-years follow-up. While in human medicine there is evidence of acquired antral stenosis, no information is available about the same lesion in the feline species. To the best of our knowledge, this is the first case of antral stenosis in cats.

Canine gastric disorders are common in veterinary clinical practice and among these neoplasms require rapid identification and characterization. Standard ultrasound (US) is the imaging modality of choice for gastric wall assessment. The aim of this prospective study is to describe the specific B-mode and contrast enhanced US (CEUS) features of normal, inflammatory, and neoplastic gastric wall in dogs. B-mode US and CEUS of the stomach were performed in anesthetized dogs with or without gastric disorders. Gastric wall qualitative and quantitative parameters were evaluated on B-mode US and CEUS examination. A total of 41 dogs were included: 6 healthy (HEA) as the control group, 9 gastritis (INF), 8 adenocarcinoma (AC), 8 alimentary lymphoma (AL), 4 leiomyosarcoma (LEIS), 2 gastrointestinal stromal tumor (GIST), 2 leiomyoma, 1 undifferentiated sarcoma, 1 metastatic gastric hemangiosarcoma. Gastric tumors appear as a marked wall thickness with absent layers definition and possible regional lymphadenopathy (AC and AL) and steatitis (AC) while gastritis generally shows no/mild thickening and no other alterations on B-mode US. On CEUS, neoplasm shows a higher and faster wash in if compared to that of gastritis. B-mode and CEUS assessment may be useful in the evaluation of canine gastric disorders in the distinction between gastritis and gastric neoplasms, even if there are no specific features able to discriminate between the different tumor histotypes.

Background and Aim: Pseudomonas aeruginosa is a relevant opportunistic and difficult to treat pathogen due to its widespread environmental diffusion, intrinsic resistance to many classes of antimicrobials, high ability to acquire additional resistance mechanisms, and wide range of pathogenic factors. The present study aimed to investigate the prevalence of P. aeruginosa in canine clinical samples, the antimicrobial susceptibility against antipseudomonal antibiotics, and the presence of extracellular pathogenic factors of the isolates, as well as their ability to produce biofilm. Materials and Methods: Overall, 300 clinical specimens from dogs with pyoderma or abscesses (n=58), otitis (n=59), and suspected bladder infection (n=183) were analyzed by standard bacteriological methods. P. aeruginosa isolates were tested for their antimicrobial susceptibility by disk and gradient diffusion methods to determine the minimum inhibitory concentrations. The ability of the isolates to produce biofilm was investigated by a microtiter plate assay, while virulence genes coding for elastase (lasB), exotoxin A (toxA), alkaline protease (aprA), hemolytic phospholipase C (plcH), and exoenzyme S (ExoS) were detected by polymerase chain reaction method. Results: A total of 24 isolates of P. aeruginosa were found in clinical specimens (urine n=3, skin/soft tissue n=6, and ear canal n=15). No resistance was found to ceftazidime, gentamicin, aztreonam, and imipenem (IMI), while low levels of resistance were found to enrofloxacin (ENR) (4.2%) and piperacillin-tazobactam (8.3%). However, 41.7% and 29.2% of the isolates showed intermediate susceptibility to ENR and IMI, respectively. Disk and gradient diffusion methods showed high concordance. The majority of the isolates revealed a weak (33.3%) or intermediate (45.8%) ability to form biofilm, while the strong biofilm producers (20.8%) derived exclusively from the ear canal samples. All isolates (100%) were positive for lasB, aprA, and plcH genes, while exoS and toxA were amplified in 21 (87.5%) and 22 (91.7%) isolates, respectively. Conclusion: In the present study, P. aeruginosa isolates from canine clinical samples were characterized by low levels of antimicrobial resistance against antipseudomonal drugs. However, the high presence of isolates with intermediate susceptibility for some categories of antibiotics, including carbapenems which are not authorized for veterinary use, could represent an early warning signal. Moreover, the presence of isolates with strong ability to produce biofilm represents a challenge for the interpretation of the antimicrobial susceptibility profile. In addition, the high prevalence of the extracellular pathogenic factors was indicative of the potential virulence of the isolates.

BackgroundThe morphological characteristics of the largest lymphatic vessels and lymph nodes of the body have been described through ultrasonography, although food and gas in the gastrointestinal tract can often have negative effects on the response of small abdominal structures. The aim of the study was to describe the size of normal abdominal lymph nodes (ALs) in dogs affected by disease, not including lymphadenomegaly or lymphadenopathy, and divided according to body weight and age. The ALs studied included the jejunal, medial iliac, portal, gastric, splenic, and pancreaticoduodenal lymph nodes.ResultsMultiple statistical analyses among the three groups showed that all measurements of the ALs increased according to body weight changes (P 0.1). Mixed results emerged from a comparison of weight categories and age; only the jejunal lymph nodes showed a possible correlation (P

A large, ill-defined, firm, multinodular mass involving the pancreas was confirmed on postmortem examination of a 5-y-old, male Rottweiler that died following acute respiratory distress syndrome, after a period of anorexia and lethargy. Histologically, the mass consisted of plump spindle cells admixed with a variable number of macrophages, lymphocytes, plasma cells, and neutrophils. Foci of coagulative necrosis and hemorrhage were also observed. Spindle cells strongly reacted to antibodies against vimentin, α–smooth muscle actin, and calponin, whereas desmin was expressed only mildly and focally. Pan-cytokeratin, KIT, glial fibrillary acidic protein, and S100 protein were nonreactive. Variable numbers of MAC 387–positive cells, CD3+ lymphocytes, and numerous blood vessels were also detected throughout the mass. Histologic and IHC findings were consistent with a diagnosis of inflammatory myofibroblastic tumor of the pancreas.

Highlights • The whole genome sequence of feline morbillivirus (FeMV) Piuma/2015 has been obtained by the combination of SISPA and NGS starting from the infected urine sample of a cat suffering of chronic kidney disease. • The highest sequence identity was detected with early FeMVs from Hong Kong. • Sequence heterogeneity exists within European FeMVs as for the existence of FeMVs in Germany and Turkey divergent from Piuma/2015., Feline morbillivirus (FeMV) has been recently identified by RT-PCR in the urine sample of a nephropathic cat in Italy. In this report, we describe the whole genome sequence of strain Piuma/2015 obtained by combination of sequence independent single primer amplification method (SISPA) and next generation sequencing (NGS) starting from RNA purified from the infected urine sample. The existence in Germany and Turkey of FeMVs from cats divergent from Piuma/2015, suggests the presence of FeMV heterogeneity in Europe as it has been described previously in Japan and China.

By screening a collection of fecal samples from young cats housed in three different shelters in South Italy, noroviruses (NoVs) were found in 3/48 (6.2%) specimens of animals with enteritis signs while they were not detected in samples collected from healthy cats (0/57). Upon sequence analysis of the short RNA-dependent RNA polymerase (RdRp) region, the three strains displayed the highest nucleotide (nt) and amino acid (aa) identities to the prototype GIV.2 strain lion/Pistoia/387/06/ITA (91.0–93.0% nt and 97.0–98.0% aa). The sequence of ~ 3.4-kb portion at the 3′ end of the genome of a NoV strain, TE/77-13/ITA, was determined. In the full-length ORF2, encoding the VP1 capsid protein, the virus was genetically closest to the canine GVI.2 NoV strains C33/Viseu/2007/PRT and FD53/2007/ITA (81.0–84.0% nt and 93.0–94.0% aa identities), suggesting a recombination nature, with the cross-over site being mapped to the ORF1-ORF2 junction. Based on the full-length VP1 amino acid sequence, we classified the novel feline NoV, together with the canine strains Viseu and FD53, as a genotype 2, within the genogroup GVI. These findings indicate that, as observed for GIV NoV, GVI strains may infect both the canine and feline host. Unrestricted circulation of NoV strains in small carnivores may provide the basis for quick genetic diversification of these viruses by recombination. Interspecies circulation of NoVs in pets must also be considered when facing outbreaks of enteric diseases in these animals., Highlights • GIV and GVI NoVs have been recently found in domestic carnivores. • In this study, NoV strains resembling animal GIV.2 NoVs in their polymerase region were detected in diarrheic cats. • One Italian strain, TE/77-13/ITA, in the full-length VP1 sequence shared the highest identity to the canine GVI.2 NoVs.

A 12-year-old female European shorthair cat was presented with severe dyspnoea. Echocardiography revealed hypertrophic cardiomyopathy and pleural effusion. The cat died from acute decompensated left heart failure. At necropsy examination, the lungs were diffusely congested and firm, with multifocal grey areas and sparse haemorrhages. No solid masses were detected. Histopathology revealed a diffuse neoplastic proliferation characterized by irregular growth along alveolar walls with a micropapillary pattern. Tumour cells were large, highly pleomorphic and intensely positive for pan-cytokeratin and CAM 5.2. Tumour growth was obscured by simultaneous lesions related to chronic pulmonary congestion and interstitial lung disease. Histological features were consistent with a diffuse invasive pulmonary adenocarcinoma with a micropapillary pattern of tumour growth. Differential diagnosis included large cell carcinoma, which is usually characterized by rosettes or solid clusters of cells occupying alveolar lumen. Extensive cytokeratin immunolabelling was helpful in the differentiation from histiocytic proliferative disease.

Objectives The aim of this study was to retrospectively describe clinical, radiographic and therapeutic features of feline lungworm infection. Methods Medical records of cats with lungworm diagnosis, thoracic radiography and without concurrent diseases between 2013 and 2015 were reviewed. Collection of data included physical examination, haematology, serum biochemistry, therapy with a variety of anthelmintics and outcomes. Results Thirty-seven records were recovered and 26 were included in the study. Single infections by Aelurostrongylus abstrusus (n = 15), Troglostrongylus brevior (n = 3) and Capillaria aerophila (n = 1) and coinfections by T brevior/ A abstrusus (n = 6) and T brevior/ C aerophila (n = 1) were diagnosed. The most common respiratory signs were coughing (n = 12), increased vesicular sounds (n = 10), dyspnoea (n = 9), such as laboured breathing, orthopnoea or open-mouth breathing, and tachypnoea (n = 6). Two cats were subclinically infected. The most common laboratory abnormality was anaemia (n = 7). Radiographic patterns recorded were interstitial (n = 24), bronchial (n = 21), alveolar (n = 10) and vascular (n = 2). Twenty-five cats had a complete recovery within 2–6 weeks of therapy. One kitten died 7 days after the diagnosis. Conclusions and relevance Lungworms should always be included in the differential diagnosis in cats living in endemic areas and presenting with respiratory signs and radiographic abnormalities. A copromicroscopic examination should be considered as the first diagnostic step for all cats at risk of lungworm infections. In most cases, timely therapy with a variety of anthelmintics guarantees recovery.

Dirofilaria immitis, Dirofilaria repens, and Acanthocheilonema reconditum are the most important canine filariae. These species are expanding their distribution in both endemic regions and in previously free areas, thus enhancing the risk for single and mixed infestations in both dogs and humans. The present study evaluated the potential use of moxidectin in a spot-on formulation in preventing these dog filarioses in a confined area where the three major canine filariae live in sympatry. The trial was performed in a private shelter with a history of mixed infestations. Twenty-nine negative dogs were monthly treated with a spot-on formulation containing moxidectin 2.5 %/imidacloprid 10 % in spring and summer of 2010. The blood of the dogs was examined monthly with classical and molecular assays until December 2011. Twenty-six dogs completed the study, and they scored constantly negative to all diagnostic assays. Three further dogs completed the study in February 2011 until when they were filariae-negative. These results show that monthly treatment with moxidectin may potentially contribute in reducing the spreading of canine filarial diseases. The permanent negativity of treated dogs for the study period is discussed in relation to the potential use of this spot-on formulation in the prevention of single and mixed infestations of major vector-borne canine filariae.

Summary The subject Dirofilaria repens and Leishmania infantum are agents of skin diseases in dogs. While features of cutaneous lesions in canine leishmaniosis are well-known, the role of D. repens in inducing dermatopathies in infected animals is still unclear. Both parasites are gaining attention, given that their geographic distribution is expanding in different European areas. Topicality and strong points The present report describes a case of simultaneous infection by L. infantum and D. repens in a dog with skin lesions. A 6-year-old dog living in Central Italy was referred for a dermatological examination, which showed an erosive-ulcerative nodular dermatitis with erythema on the inner part of left ear. The histological examination of the lesion revealed a Leishmania -induced granuloma. The skin lesion and the blood were also molecularly positive for D. repens . Circulating microfilariae were found with a value of 61 larvae/mL. Perspectives and projects The epidemiologic and sanitary importance of co-infections by D. repens and L. infantum and their possible role in causing canine dermatopathies in co-infected dogs is discussed.

In this study, the possible association between pancreatitis and treatment with methylglucamine antimoniate (MA) in dogs with leishmaniasis was evaluated. Of the 20 dogs included, three dogs showed clinical signs of pancreatitis. Abdominal ultrasound abnormalities suggestive of pancreatitis were observed in two dogs, and increased cPLI was observed in four cases. The statistical results showed a low probability of development of pancreatitis in patients treated with MA. However, considering the possible recurrence of pancreatitis in four cases in our study, we suggest that dogs treated with MA should be evaluated for pancreatitis, especially when compatible clinical signs are present.

In the past decade reports of canine subcutaneous dirofilariosis, caused by the mosquito-transmitted nematode Dirofilaria repens , increased in number in several countries in Europe, along with a rise of human cases. Given the merit to the new approaches for the control and treatment of this infection, the present study evaluated the efficacy of a single application of the spot-on formulation containing imidacloprid 10%/moxidectin 2.5% (Advocate ® , Bayer Animal Health) in the elimination of D. repens microfilariaemia in naturally infected dogs. In September 2009, 18 dogs with a natural infection by D. repens were enrolled in the study. In October 2009 all the dogs were treated once with Advocate ® and the presence/absence of circulating MF and skin lesions after treatment was evaluated monthly until April 2010. From November 2009 to April 2010 15 dogs scored negative for D. repens while one dog remained negative till March 2010 when it died. Two dogs had a recurrence of microfilariaemia in December 2009 and January 2010 respectively. Nine infected dogs showed skin lesions at the beginning of the trial, which disappeared after treatment in 7 dogs, whereas the other two symptomatic dogs did not show any dermatological improvement until the end of the trial even though they scored negative for D. repens microfilariae. This study demonstrated that a single dermal administration of Advocate ® is effective in eliminating microfilariae of D. repens and likely has a certain degree of activity in killing subcutaneous adult worms as well. This study demonstrates the efficacy of Advocate ® in the treatment of dermatitis caused by D. repens . Also, these results are of importance towards further control programs aiming to reduce the number of bites infectious for mosquitoes and the risk of infection for both humans and dogs.

Diabetes mellitus (DM) was diagnosed in a 6-year-old neutered male ferret with polyuria/polydipsia, symmetrical alopecia, and weight loss. Laboratory tests revealed severe hyperglycemia, glucosuria, and increased steroid hormone profile. Abdominal ultrasound revealed a bilateral enlargement of the adrenal glands. Significant clinical improvement was achieved with insulin- and leuprolide acetate-based therapy. After 2 months of therapy, the ferret showed a severe ketoacidosis, and the owner decided to euthanize the animal. Histological findings revealed carcinoma of the left adrenal cortex and cortical hyperplasia of the right adrenal gland. Moderate, chronic, and active pancreatitis with a marked decrease in the number of beta-cells was also present. This is the first reported case of type 1 DM associated with hyperadrenocorticism and chronic pancreatitis in a ferret.

This survey was conducted to investigate the presence of Dirofilaria immitis and Dirofilaria repens in native dogs and cats living in an area of Italy where animal and human dirofilarioses have been sporadically reported in the past few years. From May 2008 to March 2009 individual blood samples were collected from 300 native cats and 300 native dogs in the Abruzzo region of central Italy. All 600 samples were subjected to a Knott modified method, to a test for the detection of circulating D. immitis antigen and to two species-specific PCR-coupled sequencing protocols. Epidemiological data were collected and then correlated with the filarial prevalence by univariate and multivariate statistical analyses. Twenty-five dogs (8.3%) and six cats (2%) were positive for filariae in at least one diagnostic test, with an overall prevalence rate for D. immitis and D. repens of 2.3% and 5.6% in dogs and 0.3% and 1.6% in cats, respectively. Male and large sized dogs resulted more likely to be infected by Dirofilaria spp., possibly due to the fact that animals living outdoor and of large size are more exposed to mosquito bites. However these factors were not confirmed as convincing risk factors by multivariate analysis. The results are discussed, together with the origin of these new autochthonous foci in central Italy and the present expansion southward of canine and feline filariae in Italy.

Background: The diagnosis of canine diabetic ketoacidosis (DKA) usually is based on measurement of urinary acetoacetate (ketonuria). In humans, this test is less sensitive and specific than blood 3-β-hydroxybutyrate (ketonemia) evaluation. Hypothesis: Ketonemia measurement using a portable meter is more accurate than ketonuria determination with a dipstick to diagnose canine DKA. Animals: Seventy-two client-owned diabetic dogs with ketonemia, ketonuria, or both. Methods: Prospective observational study. Based on blood bicarbonate concentration and anion gap, dogs were divided into 2 groups: patients with DKA (n= 25); patients with diabetic ketosis (n= 47). Sensitivity, specificity, and positive and negative likelihood ratio (LR) at different cut-off points were determined for both ketonemia and ketonuria. Receiver operating characteristic (ROC) analysis was used to assess the accuracy of each diagnostic test to diagnose DKA. Results: With regard to ketonemia, cut-off values of 2.3 and 4.3 mmol/L revealed 100% sensitivity and 100% specificity, respectively, whereas cut-off values of 2.8 and 3.5 mmol/L showed a −LR of 0.05 and a + LR of 13.16, respectively. With regard to ketonuria, a cut-off value of 1+ revealed 92% sensitivity, 40% specificity, and −LR of 0.20, whereas a cut-off value of 3+ revealed 44% sensitivity, 94% specificity, and +LR of 6.89. The areas under the ROC curves for the ketonemia and ketonuria tests were significantly different (0.97 and 0.81, respectively, P= .003). Conclusions and Clinical Importance: Measurement of ketonemia is accurate and more effective than measurement of ketonuria to diagnose canine DKA.

L. Ragaini1*, G. Aste1, L. Cavicchioli2 and A. Boari1 1Department of Veterinary Clinical Sciences, Internal Medicine Section, School of Veterinary Medicine, University of T eramo, V.le Crispi 212, 64020, T eramo; 2Department of Public Health, Comparative Pathology and Veterinary Hygiene, School of Veterinary Medicine, University of Padua, Agripolis, V ia Romea 16, L egnaro, 35020, Padua, Italy *Correspondence: Dipartimento di Scienze Cliniche Veterinarie, Sezione di Medicina Interna, Facolta di Medicina Veterinaria, 64100 T eramo, Italy E-mail: ragaini@vet.unite.it

A. Peli1*, A. Fruganti3, G. Bettini2, G. Aste4 and A. Boari4 1Department of Veterinary Clinical Sciences, Internal Medicine Section; 2Department of Veterinary Public Health and Animal Pathology, Section of General Pathology and Anatomy Pathology, Faculty of Veterinary Medicine, University of Bologna, Bologna; 3Department of Veterinary Sciences, Section of Clinical Medicine, Faculty of Veterinary Medicine, University of Camerino, Camerino; 4Department of Clinic Veterinary Sciences, Internal Medicine Section, Faculty of Veterinary Medicine, University of T eramo, T eramo, Italy *Correspondence: Dipartimento Clinico Veterinario, Sezione di Medicina Interna, Facolta di Medicina Veterinaria, Alma Mater Studiorum, Universita di Bologna, V ia T olara di Sopra 50, 40064 Ozzano dell’Emilia (BO), Italy E-mail: apeli@vet.unibo.it