The air and dust invisible mycobiome of urban domestic environments.

Air and dust harbor a dynamic fungal biome that interacts with residential environment inhabitants usually with negative implications for human health. Fungal air and dust synthesis were investigated in houses across the Athens Metropolitan area. Active and passive culture dependent methods were employed to sample airborne and dustborne fungi for two sampling periods, one in winter and the other in summer. A core mycobiome was revealed both in air and dust constituted of the dominant Penicillium, Cladosporium, Aspergillus, Alternaria and yeasts and accompanied by several common and rare components. Penicillium and Aspergillus diversity included 22 cosmopolitan species, except the rarely found Penicillium citreonigrum, P. corylophilum, P. pagulum and Talaromyces albobiverticillius which are reported for the first time from Greece. Fungal concentrations were significantly higher during summer for both air and dust. Excessive levels of inhalable aerosol constituted mainly by certain Penicillium species were associated with indoor emission sources as these species are household molds related to food commodities rot. The ambient air fungal profile is a determinant factor of indoor fungal aerosol which subsequently shapes dustborne mycobiota. Indoor fungi can be useful bioindicators for indoor environment quality and at the same time provide insight to indoor fungal ecology.

Factors affecting the differential epithelial cell count in preputial cytology of healthy intact dogs.

Information is limited on the factors that may affect the preputial differential epithelial cell count in healthy intact dogs. Our objectives were to establish RIs of the preputial differential epithelial cell count in dogs and investigate the potential impact of signalment, presence of gross preputial discharge or neutrophils, preputial cleaning, and normal estrogen variation on the differential cell count. We enrolled prospectively 50 client-owned, healthy dogs in our study. Preputial cytology smears were obtained, and we performed a 4-part (parabasal, intermediate, superficial, cornified) differential cell count, before and after gentle preputial cleaning. We found higher counts of fully cornified cells (median: 11-12%) compared to previous studies. The cleaning procedure significantly decreased the intermediate (p = 0.002) and increased the superficial (p = 0.006) cells. Age was positively associated with intermediate (p = 0.007) and negatively with superficial (p = 0.01) and cornified (p = 0.02) cells. The concentration of estrogens was negatively associated with the percentage of parabasal cells (p = 0.04). In healthy dogs, a variable impact may be expected in the preputial differential epithelial cell count, by the cleaning of the prepuce before sampling, the age of the dogs, and the normal variation of estrogen concentration.

Bronchoscopy and Lung Fine-Needle Aspiration for Antemortem Evaluation of Pulmonary Involvement in Dogs with Naturally Occurring Canine Leishmaniosis.

Clinical manifestations from the lower respiratory tract are rare in canine leishmaniosis (CanL), making bronchoscopy and lung fine-needle aspiration (FNA) seldomly justified. The aim of this prospective study was to investigate the involvement of Leishmania infantum in the lungs of dogs with naturally occurring CanL by bronchoscopy and examination of bronchoalveolar lavage fluid (BALF), bronchial mucosa biopsies, and FNA, using immunodiagnostics. Dogs with relevant concurrent diseases and azotemia were excluded. Cough was detected in 5/31 (16.1%) dogs. Lesions (hyperemia, edema, mucosal granularity, secretions) were identified upon bronchoscopy in 19/31 (61.3%) dogs. The cytology of BALF revealed histiocytic inflammation in 14/31 (45.2%) dogs; the parasite was identified in one dog (3.2%). The immunofluorescence antibody test in BALF was positive in 15/31 (48.4%) dogs. Histopathology of bronchial mucosa and/or adjacent alveoli revealed lesions (mononuclear cell infiltration, fibrosis, edema, thickening of the inter-alveolar septa) in 24/31 (77.4%) dogs, with no Leishmania amastigotes. Positive antigen staining was observed within the cytoplasm of mononuclear cells in immunocytochemistry and immunohistochemistry. Μononuclear cells showed antigenic positivity in bronchial mucosa (27/31; 87.1%), BALF (30/31; 96.8%), and lung FNA (27/31; 87.1%). In conclusion, lungs seem to be affected from CanL more commonly than previously believed, and bronchoscopy allows obtaining valuable samples for antemortem diagnosis.

Lymphoplasmacytic Prostatitis Associated With Urethral Obstruction in a Dog; A Case Report.

A 10-year-old, male intact, mixed breed dog was admitted with a history of intermittent lower urinary tract signs of 18-months duration. Dysuria, stranguria, pollakiuria and overflow incontinence developed 2 weeks prior to presentation, while vomiting, polydipsia and anorexia were seen a few days prior to admission. Physical examination revealed prostatomegaly. Biochemistry results were compatible with postrenal azotemia. Abdominal imaging confirmed prostatomegaly with consequent urethral obstruction, and ultrasound-guided fine needle aspiration cytology of the prostate gland was consistent with granulomatous inflammation. The dog was hospitalized, urethral patency was preserved and upon laparotomy a cystostomy tube was placed, and prostatic biopsy was obtained. Orchiectomy was also performed. Urine culture performed 2 days prior to the surgery was negative. Prostatic histopathology revealed mild to moderate lymphoplasmacytic prostatitis. Three months postoperatively, clinical signs had resolved completely and urinations remained normal while the dog is still in good health 3 years after diagnosis. This is the first case of canine lymphoplasmacytic prostatitis in which clinical signs, diagnostic evaluation, management and long-term follow up are available.

Bilateral Renal Large B Cell Lymphoma in a Dog: A Case Report and Review of the Literature.

Canine lymphoma is a commonly reported neoplasia and, in most dogs, arises from lymph nodes before spreading to other organs. Renal lymphoma rarely occurs, and kidneys usually are a secondary site of origin. Primary renal lymphoma is infrequently described in the veterinary literature. In this study, we present a rare case of primary renal lymphoma in a dog and a review of similar cases. A 3-year-old male dog was admitted due to anorexia, weakness and vomiting. Clinical examination revealed bilaterally enlarged kidneys. Imaging demonstrated the presence of multiple renal masses. Cytology of abdominal fluid and kidneys led to the diagnosis of large cell lymphoma. Histopathology and immunohistochemistry on tissue samples taken from the kidneys confirmed the cytological diagnosis of lymphoma and categorized it as primary bilateral renal large B-cell lymphoma (LBCL).

Colonic mucosal and cytobrush sample cytokine mRNA expression in canine inflammatory bowel disease and their correlation with disease activity, endoscopic and histopathologic score.

Canine inflammatory bowel disease (IBD) is a group of chronic gastrointestinal disorders, the pathogenesis of which remains elusive, but it possibly involves the interaction of the intestinal immune system with luminal microbiota and food-derived antigens. Mucosal cytokines profiles in canine IBD have been investigated mainly in small intestinal disease, while data on cytokine profiles in large intestinal IBD are limited. The objective of this study was to measure colonic mucosal and cytobrush sample messenger (m)RNA expression of interleukin (IL)-1ß, IL-2, IL-12p40, IL-23p19, tumor necrosis factor-alpha (TNF-α) and chemokine C-C motif ligand (CCL28) in dogs with IBD and healthy controls using quantitative real-time polymerase chain reaction (PCR), and assess their correlation with clinical disease activity, endoscopic and histopathologic score. Dogs with IBD had a significantly increased mRNA expression of IL-1ß, IL-23p19 and CCL28 in the colonic mucosa, compared to healthy controls. None of the selected cytokines had significantly different mRNA expression in the colonic cytobrush samples between the two groups or between the colonic mucosa and cytobrush samples of dogs with IBD. Finally, there was a statistically significant correlation of clinical disease activity with endoscopic activity score and fibrosis and atrophy of the colonic mucosa in dogs with large intestinal IBD. IL-1ß, IL-23p19 and CCL28 could play a role in the pathogenesis of canine large intestinal IBD. Colonic cytokine expression does not correlate with clinical disease activity and/or endoscopic score. However, clinical signs reflect the severity of endoscopic lesions.

Colonic mucosal and serum expression of microRNAs in canine large intestinal inflammatory bowel disease.

BACKGROUND: Canine inflammatory bowel disease (IBD) is a group of chronic gastrointestinal (GI) disorders of still largely unknown etiology. Canine IBD diagnosis is time-consuming and costly as other diseases with similar signs should be initially excluded. In human IBD microRNA (miR) expression changes have been reported in GI mucosa and blood. Thus, there is a possibility that miRs may provide insight into disease pathogenesis, diagnosis and even treatment of canine IBD. The aim of this study was to determine the colonic mucosal and serum relative expression of a miRs panel in dogs with large intestinal IBD and healthy control dogs. RESULTS: Compared to healthy control dogs, dogs with large intestinal IBD showed significantly increased relative expression of miR-16, miR-21, miR-122 and miR-147 in the colonic mucosa and serum, while the relative expression of miR-185, miR-192 and miR-223 was significantly decreased. Relative expression of miR-146a was significantly increased only in the serum of dogs with large intestinal IBD. Furthermore, serum miR-192 and miR-223 relative expression correlated to disease activity and endoscopic score, respectively. CONCLUSION: Our data suggest the existence of dysregulated miRs expression patterns in canine IBD and support the potential future use of serum miRs as useful noninvasive biomarkers.

Histograms of Complete Blood Counts in Dogs: Maximizing Diagnostic Information.

Histograms, which are an integral part of the automated complete blood count, are now available through most of the automatic hematology analyzers used in veterinary clinical practice. Data concerning the size and number of blood cells are graphically presented in histograms, and their variations are also illustrated. Important information that is not apparent from numerical results are sometimes provided by histograms. Histograms are also referred to as frequency distribution curves and are essentially graphs resulting from the placement of the sizes of cells on the x-axis and the number of cells on the y-axis. Typically, automated analyzers provide histograms for each class of blood cells, that is, for erythrocytes, leukocytes, and platelets. Thus, when the erythrocyte histogram shows asymmetry with a right shift, it means the size of the erythrocytes is greater than normal (macrocytosis); when it presents a left shift, the size of the erythrocytes is less than normal (microcytosis). When two peaks are found in the curve, two populations of erythrocytes coexist, as in the case of a blood transfusion or therapeutic response. In the leukocyte histogram, three peaks are found: the closest to the y-axis (left) corresponds to the lymphocytes, the middle to the monocytes, and the right to the polymorphonuclear cells (neutrophils, eosinophils, and basophils). Finally, in platelet histogram, asymmetry with a right shift suggests the presence of giant platelets or schistocytes. Although the histogram is not recommended as a stand-alone test, it allows the practitioner to observe abnormalities in the distribution curve that correspond to abnormalities in the size or number of cells, and to quickly make diagnostic or therapeutic decisions that are particularly important in emergencies.

Evaluation of a Direct Immunofluorescent Assay and/or Conjunctival Cytology for Detection of Canine Distemper Virus Antigen.

Canine distemper is a common and potentially lethal multisystemic disease caused by the Canine distemper virus (CDV). We evaluated the diagnostic performance of direct immunofluorescent assay (FA) and cytology to detect CDV antigen in conjunctival cells compared with an established polymerase chain reaction (PCR) detection assay used as a gold standard for CDV diagnosis. Samples were collected from 57 young dogs presenting with central nervous system signs compatible with distemper disease. Exfoliative epithelial cells were collected from the right and left conjunctiva of each animal using nylon-bristled cytobrushes for cytology and cotton swabs for FA and PCR. For the direct FA, samples were stained with anti-CDV polyclonal antiserum conjugated to fluorescein isothiocyanate and imaged using a fluorescent microscope. Out of 57 dogs tested, 19 were PCR positive (15 positive in direct FA and 4 positive in cytology, including one that was negative by PCR), whereas 37 dogs were negative in all methods. A good agreement was observed between the FA and PCR, with a κ-value of 0.833 (95% CI: 0.678-0.989). Meanwhile, there was poor agreement between cytology and PCR (κ-value of 0.164; 95% CI: -0.045 to 0.373) and a fair agreement between FA and cytology (κ-value of 0.231; 95% CI: -0.026 to 0.487). Our results indicated a poor performance of cytology for the detection of CDV antigen. In contrast, FA is a 100% specific and an adequately sensitive assay (sensitivity: 78.95%, negative likelihood ratio: 0.21, 95% CI: 0.09-0.50) for antemortem diagnosis of canine distemper.

Prevalence of increased canine pancreas-specific lipase concentrations in young dogs with parvovirus enteritis.

BACKGROUND: Pancreatic abnormalities during canine parvovirus (CPV) enteritis have not been studied prospectively. OBJECTIVES: The objective of this study was to evaluate the diagnostic significance of canine serum pancreas-specific lipase (Spec cPL) concentration in dogs with CPV enteritis for the presence of acute pancreatitis (AP). Puppies with naturally occurring CPV enteritis were recruited and prospectively allocated into 2 groups according to normal or increased serum Spec cPL concentration. Clinical signs, laboratory findings, and pancreas-associated variables were compared between groups, and the impact of possible AP on disease course, duration of hospitalization, and outcome was assessed. RESULTS: Serum Spec cPL concentration in 35 puppies was above the upper limit of the RI in 17/35 (48.6%) dogs (Group A) and within the RI in 18 dogs (Group B). An increased serum lipase activity was present in 29/35 (82.9%) dogs, and Group A dogs had a higher serum lipase activity than Group B (P = .006). Serum Spec cPL in Group A dogs was positively correlated with serum lipase activity at the day of presentation (r = .667; P = .003) and day of discharge (r = .628; P = .007). No statistically significant difference was found between groups (P = .233) for the presence of systemic inflammatory response syndrome (SIRS) (6/17 or 35.3% dogs Group A, and 8/18 or 44.4% dogs Group B), the disease course, duration of hospitalization, or outcome between groups. CONCLUSIONS: Increased serum Spec cPL is relatively common in dogs with CPV enteritis. However, such increases do not seem to correlate with the outcome of disease.

Acute phase protein and antioxidant responses in dogs with experimental acute monocytic ehrlichiosis treated with rifampicin.

There is currently lack of information on the changes of acute phase proteins (APP) and antioxidant markers and their clinical relevance as treatment response indicators in canine monocytic ehrlichiosis (CME). The objective of this study was to investigate the patterns of C-reactive protein (CRP), haptoglobin (Hp), ferritin and paraoxonase-1 (PON-1) during treatment of dogs with acute CME with rifampicin. Blood serum samples from ten Beagle dogs with experimental acute CME were retrospectively examined. Five dogs (Group A) were treated with rifampicin (10mg/Kg/24h), per os, for 3 weeks and 5 dogs (Group B) received no treatment (infected controls). Two Beagle dogs served as uninfected controls. Blood serum samples were serially examined prior to Ehrlichia canis inoculation and on post-inoculation days 14, 21, 28, 35 and 42. Significant changes of CRP, Hp, ferritin and PON-1 values were found in the majority of infected dogs. However, their concentrations did not differ between the two groups during the treatment observation period. The results of this study indicate that although several APP and PON-1 tend to significantly change in the majority of dogs with acute CME, they were of limited clinical relevance as treatment response indicators in this experimental setting.

Canine coronavirus, Greece. Molecular analysis and genetic diversity characterization.

Canine coronavirus (CCoV) is an etiologic agent of diarrhea in dogs and is known to have spread worldwide. Mild disease or asymptomatic carriage are probably in many cases common outcomes of infection. To date, two different genotypes of CCoV are known, CCoV type I (CCoV-I) and CCoV type II (CCoV-II). CCoV type II is divided in two subtypes, CCoV-IIa (classical strains) and CCoV-IIb, with CCoV-IIb emerging as a result of a putative recombination between CCoV-IIa and transmissible gastroenteritis virus (TGEV). The aim of the present study was to investigate the presence of CCoV in Greece and to genetically analyze the circulating strains. Between December 2007 and December 2009, 206 fecal samples were collected from dogs with diarrhea from kennels, pet shops and veterinary clinics of different country regions. RT-PCR and real time RT-PCR assays were used for CCoV detection and characterization. CCoV was identified in 65.1% of the dogs presenting diarrhea, being more frequently detected in animals younger than 3 months old and in animals housed in groups. In 47% of the positive samples more than one CCoV genotype/subtype were detected, with triple CCoV-I/CCoV-IIa/CCoV-IIb infections being identified for the first time. Molecular and phylogenetic analysis revealed that CCoV-I Greek strains share low genetic relatedness to each other and to the prototype CCoV-I strains in the 5' end of the S gene. Moreover, a divergent CCoV-IIa strain was identified. The circulation of highly variable CCoV-I and CCoV-IIb emerging strains, as well as the detection of the divergent strain, raise concerns on the importance of these new strains as primary pathogens of diarrhoeic syndromes diagnosed in dogs.

Presumptive pure erythroid leukemia in a dog.

A 6.5-year-old, intact male Cocker Spaniel dog was referred with a history of depression and anorexia of 1-week duration. Mucosal pallor was prominent on physical examination. Complete blood cell count revealed pancytopenia and occasional blast cells. Bone marrow aspirate cytology indicated that individual particles were composed of approximately 60% hematopoietic cells and a monomorphic population of blast cells with perfectly round nuclei, consistent paranuclear clearing, and deeply basophilic cytoplasm devoid of granules dominating the marrow fields. The granulocytic lineage was severely decreased with a granulocytic-to-erythroid ratio of 0.15 and a blast cell percentage of at least 70% of all nucleated cells; the myeloblasts and monoblasts composed <5% of nonerythroid cells. Bone marrow cytology slides were submitted for immunocytochemical immunophenotyping using antibodies to myeloperoxidase, cluster of differentiation (CD)3, CD79a, CD11b, CD45, and CD34. The neoplastic cells did not express any of the antigens assessed. The combination of light microscopic cytomorphology and the immunophenotype were strongly suggestive of pure erythroid leukemia.

Canine bronchomalacia: a clinicopathological study of 18 cases diagnosed by endoscopy.

Canine bronchomalacia (BM) is characterized by weakness leading to collapse of the bronchial wall. A prospective study of 18 affected dogs (age range: 1-15 years) was undertaken to characterize the clinicopathological and histological features of BM. Poodles and Yorkshire terriers were commonly affected. Half of the dogs were overweight or obese. The clinical presentation was a mild, wheezing, chronic cough and pulmonary crackles were heard in 28% of the dogs. Compatible radiographic changes were present in 61% of the dogs. Using bronchoscopy, both lungs were affected in half of the animals, whereas in the others the disease appeared to affect predominantly the left lung. Analysis of bronchoalveolar lavage fluid and biopsies of bronchial mucosa revealed pure or mixed neutrophilic inflammation. Underlying infectious bronchitis was considered possible in 56% of the dogs. It was concluded that canine BM may present as an isolated clinical entity associated with infection and/or inflammation.

Aspergillus fumigatus Bronchopneumonia in a Hellenic Shepherd Dog.

A 3 yr old intact female Hellenic shepherd dog was referred due to depression, partial anorexia, fever, and a mild productive cough of 2 mo duration. Thoracic radiographs showed increased opacity of all of the left lung lobes. Upon bronchoscopy, a sanguineous, purulent discharge was detected in the tracheal lumen with hyperplastic tissue narrowing the left main stem bronchus. Cultures were positive for bacteria (Bacillus spp. and Clostridium spp.) but negative for fungi. Due to the severity of the lesions, a complete left lung pneumonectomy was performed. Histopathological examination of the excised lung tissues revealed a severe granulomatous bronchopneumonia with numerous alveolar macrophages laden with structures stained positively by periodic acid-Schiff and Grocott stain that had morphology consistent with fungi. PCR and sequencing of internal transcribed spacer regions 1 and 2 from genetic material extracted from paraffin-embedded pulmonary tissue confirmed the presence of Aspergillus fumigatus. Itraconazole was administrated for 5.5 mo and the dog was clinically normal 26 mo after surgery.