Hematological Parameters from the Feline Blood Donor to the Blood Unit: What Changes Are to Be Expected?

Feline blood donation requires sedation to allow for good blood collection, avoiding venous damage and unnecessary donor stress. In the present study, we describe the variation of hematological parameters in a population of 74 healthy blood-donor cats that underwent the same sedation protocol, including medetomidine, alfaxalone, and butorphanol. Changes in hematological parameters were evaluated in blood samples collected from conscious cats (group A), sedated cats (group B), and feline whole-blood units (FBUs) (group C). Significant differences were observed between unsedated and sedated cats: the mean RBC count, HGB, HCT, and RDW were significantly lower in sedated cats than in conscious ones, with a difference of 17.95%, 18.42%, 28.21%, and 10.00%, respectively. In accordance with previously reported data, our results confirm that sedation is a critical procedure in cats that can affect most of the hematological parameters. The second finding of our study is that no significant differences for the main hematological parameters were observed between sedated cats and FBUs, thus suggesting that the dilution with the conservative-anticoagulant solution exert negligible effects on these parameters with respect to samples of sedated animals. This hematological change must be taken into consideration since such parameters are important to define the quality of FBUs.

Effects of leukoreduction on storage lesions in whole blood and blood components of dogs.

BACKGROUND: Leukoreduction is a routine procedure in human transfusion medicine but is uncommon in veterinary. OBJECTIVES: To evaluate the effect of leukoreduction on the quality of canine whole blood (WB) and blood products during storage. ANIMALS: Ten canine blood donors. METHODS: This is a case series study. An amount of 450 mL of blood was collected from each dog. Five WB and 5 packed red blood cells (pRBC) bags were divided into 2 units each: leukoreduced (LR) and non-leukoreduced (nLR). RBC count, erythrocytes' mean osmotic fragility (MOF), 2,3-diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), percentage of hemolysis, potassium (K), lactate, glucose, and cytokines were measured weekly from day of donation (T0) to day 35 (T35); pH, coagulation times, and clotting factors were evaluated at T0 and T35 from WB and in fresh frozen plasma after 1 year of storage. RESULTS: Leukoreduction showed positive effects on lactate (T35: LR WB 14.42 mmol/L SD 2.71, nLR WB 22.42 mmol/L SD 1.86, LR pRBC 20.88 mmol/L SD 2.65, nLR pRBC 36.81 mmol/L SD 2.34; P < .0001), pH (T35: LR WB 6.88 SD 0.16, nLR WB 6.69 SD 0.20, P = .02; LR pRBC 6.57 SD 0.23, nLR pRBC 6.22 SD 0.11; P < .001), and K (LR pRBC 4.08 mmol/L SD 0.88, nLR pRBC 5.48 mmol/L SD 0.90; P < .001). Increasing values of IL8 were observed in nLR units during storage (T0: 4167 ± 11 888 pg/mL; T35: 6367 ± 11 612 pg/mL). CONCLUSION AND CLINICAL IMPORTANCE: LR blood units are recommended to critically ill dogs with marked inflammatory conditions.

In vitro assessment of quality of citrate-phosphate-dextrose-adenine-1 preserved feline blood collected by a commercial closed system.

BACKGROUND: Optimal procedure for storage of feline blood is needed. Open-collection systems have been employed in feline medicine, thus limiting the possibility for storage. OBJECTIVES: To evaluate indicators of quality of feline blood stored for 35 days at +4°C in a closed-collection system specifically designed for cats. ANIMALS: Eight healthy adult European domestic shorthair cats with a weight of 5-6.8 kg. METHODS: This is a case series study. A bacteriological test, CBC, blood smear, pH, osmotic fragility, 2,3-diphosphoglycerate (2,3-DPG), and adenosine triphosphate (ATP) measurement were performed weekly on whole blood (WB) units from day 1 to day 35 after donation. The hemolysis index, lactate and potassium concentrations, prothrombin time (PT), activated partial thromboplastin time (aPTT), and fibrinogen were measured on plasma aliquots. RESULTS: One out of eight blood units (BUs) had bacterial growth (Serratia marcescens) at day 35. No significant differences were found regarding CBC, morphology, pH, and osmotic fragility. Despite high inter-individual variability and low starting levels, significant decreases in the mean concentrations of 2,3-DPG (T0 1.99 mmol/g Hb, SD 0.52, T35 1.25 mmol/g Hb, SD 1.43; P = .003) and ATP (T0 1.45 mmol/g Hb, SD 0.71, T35 0.62 mmol/g Hb, SD 0.51; P < .001) were detected during the study, as opposed to an increase in hemolysis (T0 0.11 mmol/L, SD 0.07, T35 0.84 mmol/L, SD 0.19; P < .001), lactate (T0 3.30 mmol/L, SD 0.86, T35 13.36 mmol/L, SD 2.90; P < .001), and potassium (T0 3.10 mmol/L, SD 0.21, T35 4.12 mmol/L, SD 0.35; P < .001) concentrations. CONCLUSIONS AND CLINICAL IMPORTANCE: The commercial BU kit is appropriate for blood collection and conservation of WB in cats. The maintenance of WB quality indicators during storage is essential for future improvements of feline transfusion medicine.

DNA and RNA isolation from canine oncologic formalin-fixed, paraffin-embedded tissues for downstream "-omic" analyses: possible or not?

Formalin-fixed, paraffin-embedded (FFPE) tissues represent a unique source of archived biological material, but obtaining suitable DNA and RNA for retrospective "-omic" investigations is still challenging. In the current study, canine tumor FFPE blocks were used to 1) compare common commercial DNA and RNA extraction kits; 2) compare target gene expression measured in FFPE blocks and biopsies stored in a commercial storage reagent; 3) assess the impact of fixation time; and 4) perform biomolecular investigations on archival samples chosen according to formalin fixation times. Nucleic acids yield and quality were determined by spectrophotometer and capillary electrophoresis, respectively. Quantitative real-time polymerase chain reaction assays for the following genes: BCL-2-associated X protein, B-cell lymphoma extra large, antigen identified by monoclonal antibody Ki-67, proto-oncogene c-KIT (c-kit). Two internal control genes (Golgin A1 and canine transmembrane BAX inhibitor motif containing 4), together with direct sequencing of c-kit exons 8, 9, 11, and 17, were used as end points. Differences in DNA/RNA yield and purity were noticed among the commercial kits. Nucleic acids (particularly RNA) extracted from paraffin blocks were degraded, even at lower fixation times. Compared to samples held in the commercial storage reagent, archived tissues showed a poorer amplification. Therefore, a gold standard protocol for DNA/RNA isolation from canine tumor FFPE blocks for molecular investigations is still troublesome. More standardized storage conditions, including time between sample acquisition and fixation, fixation time, and sample thickness, are needed to guarantee the preservation of nucleic acids and, then, their possible use in retrospective transcriptomic analysis.

Adenocarcinoma of the dorsal glands in 2 European ground squirrels (Spermophilus citellus).

Olfactory communication is an important aspect of the biology of ground squirrels; accordingly, some of their integumentary glands are associated with scent-marking behavior. Although reports of neoplasms in ground squirrels are limited, the literature on tumors in this family of rodents is extensive, with hepatocellular carcinomas in woodchucks and fibromas in squirrels being the 2 most common neoplasms. Apocrine gland tumors occur frequently in domestic animals such as cats and dogs but to our knowledge have not previously been reported in squirrels. Here we describe 2 cases of adenocarcinoma of the dorsal glands in privately owned European ground squirrels (Spermophilus citellus). The skin nodules were characterized histologically by proliferation of epithelial cells, which were arranged in a tubuloacinar pattern with neoplastic emboli within the blood vessels. Adenocarcinoma of the dorsal glands was diagnosed in light of the anatomic localization, immunohistochemistry results, and histochemistry findings.

Retroperitoneal teratoma in a domestic rabbit (Oryctolagus cuniculus).

Teratomas are histologically complex tumors arising from germ cells of two or three embryonic layers either in the gonads or occasionally from germ cells that mistakenly migrated to ectopic sites. A case of retroperitoneal teratoma in a regularly slaughtered 4-month-old, male domestic rabbit is reported. A pale reddish, 3 x 4 cm, fibrous mass was detected, loosely connected to the parietal wall in the retroperitoneal region cranial to the kidney. Histologically, the neoplasm contained tissues from ectodermal, endodermal and mesodermal germ layers: nerves and neurons simulating ganglia, cysts lined by squamous epithelium containing keratin and hair, skin adnexa; respiratory glandular epithelium, serous and salivary glands; collagenous connective tissue, muscle fibers, mature cartilage and bone, bone marrow, teeth and fat. To the authors' knowledge this is the first report of a retroperitoneal teratoma in the rabbit.

Intra-abdominal leiomyosarcoma in a ferret (Mustela putorius Furo): histopathological and immunohistochemical characterization.

A 5 years old female ferret with an abdominal palpable mass confirmed at echo-graphic examination died during an explorative laparotomy. A single lymph-node-like nodule was found adjacent to the intestinal loops. The round mass well circumscribed, solid and white, histologically, at low magnification, appeared encapsulated and built up by a population of atypical spindle cells arranged in interwoven fascicles. The cells had high anisocytosis degree, moderate mitotic activity and prominent nucleoli. A central area of necrosis was present. To characterise the tumour immunohistochemically cytokeratin, vimentin, S-100, melan-A, vWF, desmin, actin and alpha-actin were applied. Neoplastic cells resulted positive to vimentin, actin and alpha-actin. Based on the histological and immunohistological pattern a diagnosis of leiomyosarcoma was made.

Unilateral seminoma with multiple visceral metastases in a duck (Anas platyrhynchos).

A case of unilateral seminoma with visceral metastases in a Mallard duck (Anas platyrhynchos) is reported. The right testis was markedly enlarged. The liver surface showed multifocal to coalescent regular circular umbilicated greyish-white spots. In addition, multiple rough whitish nodules were evident on the pancreas and the visceral peritoneum lining the intestine. Histologically, the right testicular parenchyma was diffusely affected and replaced by neoplastic growth, consisting of sheets of large round to polyhedral cells with conspicuous vesicular nuclei having distinctly granular chromatin and prominent nucleoli. Sheets of cells with similar features were observed also in the other affected organs. Multiple lung metastases were detected on histology. This is the first known report of seminoma with hepatic, pancreatic, pulmonary and peritoneal metastases in a Mallard duck.