Effect of leukoreduction on the omics phenotypes of canine packed red blood cells during refrigerated storage.

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.

Effect of leukoreduction on the metabolism of equine packed red blood cells during refrigerated storage.

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.

ZOOMICS : comparative metabolomics of red blood cells from dogs, cows, horses and donkeys during refrigerated storage for up to 42 days.

BACKGROUND: The use of omics technologies in human transfusion medicine has improved our understanding of the red blood cell (RBC) storage lesion(s). Despite significant progress towards understanding the storage lesion(s) of human RBCs, a comparison of basal and post-storage RBC metabolism across multiple species using omics technologies has not yet been reported, and is the focus of this study. MATERIALS AND METHODS: Blood was collected in a standard bag system (CPD-SAG-Mannitol) from dogs (n=8), horses, bovines, and donkeys (n=6). All bags were stored at 4°C for up to 42 days (i.e., the end of the shelf life in Italian veterinary clinics) and sampled weekly for metabolomics analyses. In addition, data comparisons to our ongoing Zoomics project are included to compare this study's results with those of non-human primates and humans. RESULTS: Significant interspecies differences in RBC metabolism were observed at baseline, at the time of donation, with bovine showing significantly higher levels of metabolites in the tryptophan/kynurenine pathway; dogs showing elevated levels of high-energy compounds (especially adenosine triphosphate and S-adenosyl-methionine) and equine (donkey and horse) RBCs showing almost overlapping phenotypes, with the highest levels of free branched chain amino acids, glycolytic metabolites (including 2,3-diphosphoglycerate), higher total glutathione pools, and elevated metabolites of the folate pathway compared to the other species. Strikingly, previously described metabolic markers of the storage lesion(s) in humans followed similar trends across all species, though the rate of accumulation/depletion of metabolites in energy and redox metabolism varied by species, with equine blood showing the lowest degree of storage lesion(s). DISCUSSION: These results interrogate RBC metabolism across a range of mammalian species and improve our understanding of both human and veterinary blood storage and transfusion.

Cytauxzoon sp. Infection and Coinfections in Three Domestic Cats in Central Italy.

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.

Detection of Serum Allergen-Specific IgE in Atopic Dogs Tested in Northern Italy: Preliminary Study.

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.

Frequency of Blood Types A, B and AB in a Population of Non-Pedigree Domestic Cats from Central Italy.

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.

Invasive pleural malignant mesothelioma with rib destruction and concurrent osteosarcoma in a dog.

A 7-year-old Dachshund was clinically examined because of a 10-day history of lameness in the left hind limb. On the basis of radiological and cytological findings, an osteosarcoma of the left acetabular region was suspected. The dog underwent a hemipelvectomy and osteosarcoma was diagnosed by subsequent histopathological examination. An immovable subcutaneous mass was noted on the left chest wall during the physical examination and non-septic neutrophilic inflammation was diagnosed by cytology. Forty days later, the dog showed signs of respiratory distress with an in-diameter increase of the subcutaneous mass up to 4 cm. Thoracic radiography and ultrasonography revealed pleural effusion and a lytic process in the fourth left rib. Furthermore, ultrasound examination revealed a mixed echogenic mobile structure with a diameter of around 2 cm floating within the pleural fluid of the left hemithorax close to the pericardium. The dog underwent surgery for an en bloc resection of the subcutaneous mass together with the fourth rib and the parietal pleura. Moreover, the left altered lung lobe, corresponding to the mobile structure detected by ultrasound, was removed. Based on cytological, histopathological, and immunohistochemical examinations, an invasive epithelioid pleural malignant mesothelioma was diagnosed.

Allergic dermatitis by Dirofilaria repens in a dog: clinical picture and treatment.

Adult stages of Dirofilaria repens (Nematoda, Filarioidea) reside in the subcutaneous tissues of the definitive or occasional host as dogs, other animals, and humans, and it is transmitted by mosquitoes. Canine infections with adults and circulating larvae of D. repens are often considered asymptomatic, although in some cases, the parasite causes subcutaneous nodules, diffused dermatitis, skin lesions, and itching. This report provides a complete clinical description of an unusual case of allergic diffused dermatitis caused by D. repens in a naturally infected dog and its successful treatment with the use of a spot-on solution containing imidacloprid 10%/moxidectin 2.5%. The dog presented multiple pustules and alopecic areas with lichenification, hyperpigmentation, and erythematous scaling margins without pruritus. Histological examination was compatible with allergic dermatitis. After being unsuccessfully managed for suspected food hypersensitivity, with a significantly worsening of the lesions, a Knott's analysis detected nematode larvae in the blood. Morphological and molecular identification showed them to be D. repens. The dog was then treated with a single administration of a spot-on formulation containing imidacloprid 10%/moxidectin 2.5%, and the dermatological signs completely resolved within 2 months after treatment. The dog showed no recurrence of the lesions, and no circulating microfilariae were found upon microscopic and molecular examination for six consecutive months after treatment. This report indicates the apparent primary role of D. repens in causing hypersensitivity-like skin disease without pruritus in a dog. It also confirms, as recently shown elsewhere, the efficacy of imidacloprid 10%/moxidectin 2.5% in the treatment of dermatitis caused by D. repens.