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Abstract Background Red blood cell (RBC) storage promotes biochemical and morphological alterations, collectively referred to as storage lesions (SLs). Studies in humans have identified leukoreduction (LR) as a critical processing step that mitigates SLs. To date no study has evaluated the impact of LR on metabolic SLs in canine blood units using omics technologies. Objective Compare the lipid and metabolic profiles of canine packed RBC (pRBC) units as a function of LR in fresh and stored refrigerated (up to 42 days) units. Animals Packed RBC units were obtained from 8 donor dogs enrolled at 2 different Italian veterinary blood banks. Study Design and Methods Observational study. A volume of 450 mL of whole blood was collected using Citrate‐Phosphate‐Dextrose‐Saline‐Adenine‐Glucose‐Mannitol (CPD‐SAGM) transfusion bags with a LR filter to produce 2 pRBC units for each donor, without (nLR‐pRBC) and with (LR‐pRBC) LR. Units were stored in the blood bank at 4 ± 2°C. Sterile weekly samples were obtained from each unit for omics analyses. Results A significant effect of LR on fresh and stored RBC metabolic phenotypes was observed. The nLR‐pRBC were characterized by higher concentrations of free short and medium‐chain fatty acids, carboxylic acids (pyruvate, lactate), and amino acids (arginine, cystine). The LR‐pRBC had higher concentrations of glycolytic metabolites, high energy phosphate compounds (adenosine triphosphate [ATP]), and antioxidant metabolites (pentose phosphate, total glutathione). Conclusion and Clinical Importance Leukoreduction decreases the metabolic SLs of canine pRBC by preserving energy metabolism and preventing oxidative lesions.

Abstract Background Understanding of the biochemical and morphological lesions associated with storage of equine blood is limited. Objective To demonstrate the temporal sequences of lipid and metabolic profiles of equine fresh and stored (up to 42 days) and leukoreduced packed red blood cells (LR‐pRBC) and non‐leukoreduced packed RBC (nLR‐pRBC). Animals Packed RBC units were obtained from 6 healthy blood donor horses enrolled in 2 blood banks. Methods Observational study. Whole blood was collected from each donor using transfusion bags with a LR filter. Leukoreduction pRBC and nLR‐pRBC units were obtained and stored at 4°C for up 42 days. Sterile weekly sampling was performed from each unit for analyses. Results Red blood cells and supernatants progressively accumulated lactate products while high‐energy phosphate compounds (adenosine triphosphate and 2,3‐Diphosphoglycerate) declined. Hypoxanthine, xanthine, and free fatty acids accumulated in stored RBC and supernatants. These lesions were exacerbated in non‐LR‐pRBC. Conclusion and Clinical Importance Leukoreduction has a beneficial effect on RBC energy and redox metabolism of equine pRBC and the onset and severity of the metabolic storage lesions RBC.

Chronic enteropathies (CEs) in dogs, according to the treatment response to consecutive trials, are classified as food-responsive (FRE), antibiotic-responsive (ARE), and immunosuppressive-responsive (IRE) enteropathy. In addition to this classification, dogs with loss of protein across the gut are grouped as protein-losing enteropathy (PLE). At present, the diagnosis of CEs is time-consuming, costly and sometimes invasive, also because non-invasive biomarkers with high sensitivity and specificity are not yet available. Therefore, this study aimed at assessing the levels of circulating endocannabinoids in plasma as potential diagnostic markers of canine CEs. Thirty-three dogs with primary chronic gastrointestinal signs presented to Veterinary Teaching Hospitals of Teramo and Bologna (Italy) were prospectively enrolled in the study, and 30 healthy dogs were included as a control group. Plasma levels of N-arachidonoylethanolamine (AEA), 2-arachidonoylglycerol (2-AG), N-palmitoylethanolamine (PEA), and N-oleoylethanolamine (OEA) were measured at the time of the first visit in dogs with different CEs, as well as in healthy subjects. Plasma levels of 2-AG (p = 0.001) and PEA (p = 0.008) were increased in canine CEs compared to healthy dogs. In particular, PEA levels were increased in the FRE group compared to healthy dogs (p = 0.04), while 2-AG was higher in IRE than in healthy dogs (p = 0.0001). Dogs affected by FRE also showed decreased 2-AG (p = 0.0001) and increased OEA levels (p = 0.0018) compared to IRE dogs. Moreover, dogs with PLE showed increased 2-AG (p = 0.033) and decreased AEA (p = 0.035), OEA (p = 0.016) and PEA (p = 0.023) levels, when compared to dogs affected by CEs without loss of proteins. The areas under ROC curves for circulating 2-AG (0.91; 95% confidence interval [CI], 0.79–1.03) and OEA (0.81; 95% CI, 0.65–0.97) showed a good accuracy in distinguishing the different forms of CEs under study (FRE, ARE and IRE), at the time of the first visit. The present study demonstrated that endocannabinoid signaling is altered in canine CEs, and that CE subtypes showed distinct profiles of 2-AG, PEA and OEA plasma levels, suggesting that these circulating bioactive lipids might have the potential to become candidate biomarkers for canine CEs.

Molecular-based approaches are rapidly developing in medicine for the evaluation of physiological and pathological conditions and discovery of new biomarkers in prevention and therapy. Fatty acid diversity and roles in health and disease in humans are topical subjects of lipidomics. In particular, membrane fatty acid-based lipidomics provides molecular data of relevance in the study of human chronic diseases, connecting metabolic, and nutritional aspects to health conditions. In veterinary medicine, membrane lipidomics, and fatty acid profiles have not been developed yet in nutritional approaches to health and in disease conditions. Using a protocol widely tested in human profiling, in the present study erythrocyte membrane lipidome was examined in 68 clinically healthy dogs, with different ages, sex, and sizes. In particular, a cluster composed of 10 fatty acids, present in membrane glycerophospholipids and representative of structural and functional properties of cell membrane, was chosen, and quantitatively analyzed. The interval values and distribution for each fatty acid of the cluster were determined, providing the first panel describing the healthy dog lipidomic membrane profile, with interesting correlation to bodyweight increases. This molecular information can be advantageously developed as benchmark in veterinary medicine for the evaluation of metabolic and nutritional status in healthy and diseased dogs.

The placement of peripheral intravenous catheters (PIVC) is potentially associated with complications that negatively impact healthcare. Our study investigated factors associated with the occurrence of PIVC-related complications in dogs and cats at a Veterinary Teaching Hospital. The second aim was to determine the prevalence of PIVC bacterial colonization. A total of 76 dogs and 40 cats with PIVCs were evaluated for the occurrence of phlebitis and mechanical complications. The devices were removed when they ceased to be functional or when complications occurred, and the content was submitted for bacterial cultures and antimicrobial susceptibility tests. Both multivariable linear regression models and ROC analysis were employed. Complications were recorded in 46.6% of cases, and 20.7% of catheters yielded a positive culture. Among the isolates, 45% were classified as multi-resistant. In dogs, a ≥36-h indwelling time was associated with an increased risk of complications. Male cats seem more prone to developing complications, while the insertion of PIVCs under sedation may represent a protective factor in this species. In conclusion, PIVC-associated complications were frequently observed, and the high rate of positive culture for PIVCs, together with the presence of multi-resistant isolates, is a cause of concern in a hospital setting.

Cytauxzoonosis is an emerging disease caused by a tick-transmitted haemoprotozoan affecting domestic and wild felids. The clinical and biomolecular findings of the infection due to Cytauxzoon sp. and concomitant coinfections are described in three cats in central Italy. Three domestic cats were referred for different clinical conditions (impact trauma, lameness, and weight loss and lethargy). They presented different hematobiochemical profiles. Only two cats were anemic, but in all three cats, endo erythrocyte inclusions suggestive of piroplasmids were found at blood smear evaluation. EDTA blood samples were submitted to rapid ELISA test for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV), and to biomolecular investigations for Piroplasmida (Babesia spp., Theileria spp., Cytauxzoon spp.) and Mycoplasma spp. All three cats were positive for Cytauxzoon sp. (European Cytauxzoon species) and two cases were also coinfected by Candidatus Mycoplasma turicensis and FIV. This report suggests that cytauxzoonosis should be included among differential diagnoses in subjects with possibility of contact with ticks and with presence of coinfections by tick-borne parasites, including in non-endemic areas.

Canine chronic enteropathies (CEs) are inflammatory processes resulting from complex interplay between the mucosal immune system, intestinal microbiome, and dietary components in susceptible dogs. Fatty acids (FAs) play important roles in the regulation of physiologic and metabolic pathways and their role in inflammation seems to be dual, as they exhibit pro–inflammatory and anti–inflammatory functions. Analysis of red blood cell (RBC) membrane fatty acid profile represents a tool for assessing the quantity and quality of structural and functional molecular components. This study was aimed at comparing the FA membrane profile, determined by Gas Chromatography and relevant lipid parameter of 48 CE dogs compared with 68 healthy dogs. In CE patients, the levels of stearic (p < 0.0001), dihomo–gamma–linolenic, eicosapentaenoic (p = 0.02), and docosahexaenoic (p = 0.02) acids were significantly higher, and those of palmitic (p < 0.0001) and linoleic (p = 0.0006) acids were significantly lower. Non-responder dogs presented higher percentages of vaccenic acid (p = 0.007), compared to those of dogs that responded to diagnostic trials. These results suggest that lipidomic status may reflect the “gut health”, and the non–invasive analysis of RBC membrane might have the potential to become a candidate biomarker in the evaluation of dogs affected by CE.

The cat lungworm Aelurostrongylus abstrusus is a cause of lower respiratory tract disease worldwide. Bronchoscopy and bronchoalveolar lavage (BAL) are important tools for diagnosing respiratory diseases in cats. Therefore, the aim of the study was to investigate the usefulness of bronchoscopy and BAL in the diagnosis of A. abstrusus. Findings from bronchoscopic examination and BAL of 24 naturally infected cats were evaluated and compared with those of 12 cats with idiopathic Feline Bronchial Diseases (FBDs). Data were analyzed using Mann-Whitney or Fisher's exact tests. No significant bronchoscopic differences were detected between cats with aelurostrongylosis and FBDs in bronchial mucus, nodular lesions, and airway collapse. On the other hand, airway hyperemia, epithelial irregularities, and bronchial stenosis were observed more frequently in cats affected by FBDs than aelurostrongylosis, while bronchiectasis was found only in cats infected by A. abstrusus. Neutrophilic, eosinophilic, lymphocytic, and mixed inflammation were recorded in both groups. Bacteria or bacterial DNA was identified regardless of the presence or absence of A. abstrusus with no significant differences between groups. Larvae of A. abstrusus were cytologically detected in 5 of the 24 cats (20.8%) with aelurostrongylosis. These results indicate that, although some findings on bronchoscopic examination (i.e., bronchiectasis) can be described more frequently in cats infected by A. abstrusus, bronchial alterations and cytological findings in aelurostrongylosis are not specific unless larvae are observed and overlap with those of other feline airway diseases.

Diagnosis of thoracic lesions on the basis of history and physical examination is often challenging. Diagnostic imaging is therefore of paramount importance in this field. Radiology has traditionally been considered the diagnostic procedure of choice for these diseases. Nevertheless, it is often not possible to differentiate inflammatory/infectious lesions from neoplastic diseases. A correct cytological and histopathologic diagnosis is therefore needed for an accurate diagnosis and subsequent prognostic and therapeutic approach. In human medicine, Computed Tomography (CT) and CT-guided biopsy are used in the presence of lesions which are not adequately diagnosed with other procedures. In the present study, thoracic lesions from 52 dogs and 10 cats of different sex, breed and size underwent both CT-guided fine-needle aspiration (FNAB) and tissue-core biopsy (TCB). Clinical examination, hematobiochemical analysis and chest radiography were performed on all animals. In this study, 59 of 62 histopathological samples were diagnostic (95.2%). Cytology was diagnostic in 43 of 62 samples (69.4%). General sensitivity, accuracy and PPV for FNAB and TCB were 67.7%, 67.7% and 100% and 96.7%, 95.2% and 98.3%, respectively. Combining the two techniques, the overall mean accuracy for diagnosis was 98.4%. Nineteen of 62 cases showed complications (30.6%). Mild pneumothorax was seen in 16 cases, whereas mild hemorrhage occurred in three cases. No major complications were encountered. CT-guided FNAB cytology can be considered a useful and reliable technique, especially for small lesions or lesions located close to vital organs and therefore dangerous to biopsy in other way.

Canine atopic dermatitis (CAD) is a pruritic allergic skin disease associated with IgE-mediated hypersensitivity. IgE is detected using Serum Allergen-Specific IgE test (SAT) in order to identify allergens. The present study aims to identify the environmental allergens in atopic dogs living in Northern Italy using SAT. The screening SAT (sSAT), using a monoclonal antibody cocktail-based ELISA to identify indoor and outdoor allergens, was performed. In all positive samples, an anti-IgE monoclonal antibody ELISA test was performed to extend panel of allergens. Out of 117 selected dogs, 69 were included in the study; 71% were positive and 29% were negative to sSAT. Among the 49 positive sSAT, 53% were positive for both indoor and outdoor, 38.8% only for indoor, and 8.2% only for outdoor allergens. This is the first study on the frequency of allergens involved in CAD in Italy using SAT. IgE hypersensitivity in atopic dogs of Northern Italy is usually associated with indoor allergens, primarily house dust mites. Among the outdoor allergens, an important role was played by Rumex acetosa. Polysensitization also commonly occurs. Therefore, since the numerous factors affect the IgE positivity in CAD, specific panels for geographical areas should be considered and re-evaluated at time intervals.

The liver has a unique vascular supply, and triple-phase contrast-enhanced computed tomography examinations are being performed in order to characterize liver lesions. This study aimed to look for any associations between the attenuation values of liver lesions and their histological classification. The inclusion criteria for this retrospective study were focal or multifocal liver lesions and histological diagnosis. All of the dogs underwent pre-contrast and triple-phase postcontrast computed tomography (CT) examinations with identical timings of the postcontrast series. Thirty-one dogs were included in the study, and various benign and malignant pathologies were identified. The results did not identify any significant differences between the benign and malignant liver lesions, nor between the individual histological diagnoses. Inflammatory lesions were significantly different compared to the normal liver parenchyma, and significant hypoattenuation was found in the portal and delayed venous phases. Hemangiosarcomas were significantly hypoattenuating to the normal liver parenchyma in the pre-contrast and arterial phases, and also to all of the benign lesions in the arterial phase. The other pathologies showed variable attenuation patterns in the different postcontrast phases, and differentiation was not possible. On the basis of this study, triple-phase contrast-enhanced computed tomography cannot differentiate between benign and malignant liver lesions, and biopsy and further histological analysis are necessary.

Blood transfusion reactions and neonatal isoerythrolysis are common events in the feline population due to the presence of natural alloantibodies in the AB blood group system. It is known that the frequency of feline blood types varies according to the geographic region and breed. Therefore, the aims of this study were to investigate the frequency of AB blood groups in non-pedigree domestic cats in Central Italy and estimate the risk of a life-threatening transfusion reaction and neonatal isoerythrolysis, caused by mismatched transfusion or incompatible random mating, respectively. The AB blood group was determined on non-pedigree domestic feline patients and potential blood donors submitted at the Veterinary Teaching Hospitals of the Universities of Teramo (Abruzzo Region, Teramo, Italy) and Perugia (Umbria Region, Teramo, Italy), and visited at veterinary practitioners in Rome (Lazio Region, Teramo, Italy) using commercial immunochromatographic cartridges and commercial agglutination cards. There were four hundred and eighty-three cats included in the study. The frequencies of the blood types were: 89.9% type A, 7.0% type B, and 3.1% type AB. The probability of an acute hemolytic transfusion reaction or a neonatal isoerythrolysis was 6.5%. Although the frequency of type B in non-pedigree domestic cats living in Central Italy was relatively low, to reduce the risk of fatal transfusion reactions, blood group typing is recommended before each transfusion.

Feline morbillivirus (FeMV) is a newly discovered paramyxovirus infecting domestic cats and its role in the pathogenesis of feline chronic kidney disease (CKD) has been suggested, however not confirmed. The primary aim of the study was to evaluate the renal damage associated with FeMV infection in cats. In this retrospective study, clinical and clinicopathological data were compared among 14 FeMV naturally infected, 21 CKD and 22 healthy cats. FeMV positive cats had serum chemistry analytes and main urine chemistry results similar to the healthy subjects. FeMV positive cats had significantly decreased urine specific gravity (median 1054, range 1022–1065) and urine creatinine (median 227.23 mg/dL, range 83.02–489.75) when compared with healthy cats (median 1067, range 1040–1080, p < 0.001; median 406.50 mg/dL, range 195.32–575.58; p < 0.001, respectively). Urine protein:creatinine ratio (UPC) results of FeMV and CKD were not different (median 0.20, range 0.08–1.03; median 0.23, range 0.10–0.80, respectively), however UPC results were significantly increased in both groups, if compared with healthy cats (median 0.1, range 0.04–0.250, p < 0.01). Based on clinical data, serum creatinine concentration, urine specific gravity and UPC results, CKD was suspected by clinicians in 3/14 FeMV cats. Urine protein sodium-dodecyl-sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in 10/13 (77%) FeMV cats indicated a tubular pattern, with a decrease of uromodulin and an increase in the number and intensity of low molecular weight proteins. FeMV infection can be associated with different grades of renal dysfunction ranging from mild tubular proteinuria with less concentrated urine to azotemia in cats younger than those typically affected by CKD.