BackgroundThe role of inflammation in the development and progression of chronic kidney disease (CKD) in cats is not well characterized. Hepcidin is a recently discovered acute‐phase protein (APP) that plays an important role in iron metabolism and contributes to the development of anemia in humans with CKD.ObjectivesTo compare serum APP concentrations, iron status, and erythropoietin (EPO) concentrations in healthy cats and cats with naturally occurring CKD.AnimalsA total of 18 healthy control cats and 38 cats with CKD.MethodsProspective study. After complete physical examination and routine blood analysis, the following tests were performed: serum amyloid A (SAA), haptoglobin (HAP), EPO, serum iron and ferritin concentration as well as total iron‐binding capacity (TIBC). Serum hepcidin‐25 concentration was measured by ELISA kit designed for use in humans.ResultsMean SAA and hepcidin concentrations were significantly higher and mean total iron and TIBC were significantly lower in the CKD group (P < .05). There was a significant positive correlation between serum creatinine concentration (CRT) and 2 of the APPs (SAA and hepcidin; P < .05). Increases in SAA and hepcidin were associated with decreases in TIBC and hematocrit in the CKD group. Fourteen (37%) of the cats with CKD were anemic, and these cats had significantly lower TIBC (P < .05), suggesting a functional iron deficiency. There was no association between survival time and APP, iron status, or EPO concentrations.ConclusionsOur data suggest that CKD in cats is associated with systemic inflammation and altered iron metabolism. With further validation in cats, hepcidin assays may help better characterize these relationships.

| INTRODUC TI ONThe CBC is an essential component of patient health evaluations.Automated cell counters have been used since the mid-20th century 1 to rapidly obtain certain hematology data, including several red blood cell (RBC), white blood cell (WBC), and platelet (PLT) indices. However, blood smear evaluation remains a requisite part of the CBC in veterinary species, as automated analyzers do not reliably detect many clinically relevant morphologic abnormalities, such as immature granulocytes, toxic change, infectious agents, and neoplastic cells. Evaluation and, therefore, interpretation of CBC findings can be limited by certain preanalytical factors. For example, prolonged storage could cause artifactual increases in hematocrit, MCV, and MPV, and decreases in the MCHC and PLT variables. 2-6 Additionally, in 1984, Gossett and Carakostas 7 showed that Abstract Background: The presence of toxic change in neutrophils is frequently used as a biomarker of inflammation in dogs. Objective: We aimed to evaluate the effect of time and storage on toxic change in canine neutrophils. Methods: One hundred and fifty microliters of EDTA blood were obtained from eight dogs with no toxic neutrophil changes observed on fresh blood smears (T0). Blood was stored at room temperature (RT), in a box with an icepack (ICE), and at 4°C. For each storage condition, smears were prepared 2 (T2), 4 (T4), 8 (T8), and 24 (T24) hours post blood draw. Smears were randomized, and each smear was evaluated for the presence of toxic neutrophil change. Results: A statistically significant effect of time and storage on the presence of toxic neutrophil change was observed. Compared with T0, the number of neutrophils containing Döhle bodies was significantly higher at T8 and T24 for the RT (P < 0.0001) and ICE (P < 0.0001) samples and at T24 for 4°C samples (P < 0.0001). Additionally, smears were falsely classified as having 1+ toxic change in 0/8 (T2), 1/8 (T4), 3/8 (T8), and 8/8 (T24) for RT samples; 0/8 (T2 and T4), 2/8 (T8), and 5/7 (T24) smears for ICE samples; and 0/8 (T2, T4, and T8) and 2/8 (T24) for 4°C samples. Conclusions: Smears can be falsely classified as having neutrophils with toxic change as early as 4 hours post blood draw in samples stored at RT, 8 hours when stored with icepacks, and 24 hours when stored at 4°C. Canine blood smears should be prepared and evaluated for toxic neutrophil change as early as possible. K E Y W O R D S artifact, delay, Döhle bodies, icepack, preanalytic, refrigerator

Glucose and trehalose are the main energy sources used by honeybees ( Apis mellifera) for daily activities. However, there is no validated point-of-care method to reliably measure both sugars. We performed an analytical validation of a portable human glucometer (Accu-Chek; Roche) for glucose measurement in honeybee hemolymph compared to a reference method (GluCH, UniCel DxC 600; Beckman Coulter). We used 30 pooled hemolymph samples collected from the antennae of anesthetized honeybees and diluted 1:4 in 0.9% saline. We evaluated dilution linearity, spike recovery, and inter- and intra-assay imprecision. Glucose concentration was measured over time (2 h, 4 h, 8 h, 12 h, 1 d, 2 d, 3 d, 7 d, 21 d, 28 d) at various storage temperature (25°C, 4°C, −20°C, −80°C). The trehalose concentration was measured indirectly by trehalase hydrolyzation. Glucose concentrations measured by both instruments had a strong correlation (0.985, p < 0.0001) and a bias of −7.33 mmol/L (±1.96SD: 13.70 to −28.36), with linear agreement at <20 mmol/L (physiologic value: 100 mmol/L). The accuracy of the glucometer decreased at >20 mmol/L. Recovery of 115–130% of diluted spikes indicated good specificity. Inter- and intra-assay imprecision were 2.50% and 2.21%, respectively. Glucose concentrations fluctuated in stored samples dependent on time and temperature; however, glucose concentrations were constant with storage at −80°C for ≥28 d. The Accu-Chek glucometer is an adequate instrument to measure honeybee glucose concentration in hemolymph diluted with 0.9% NaCl, with good accuracy and precision at <20 mmol/L. Hemolymph storage at −80°C is suitable for long-term conservation of glucose.

The European honeybee contributes to the agriculture by its pollination; however, the overwintering loss rate over the last decades is worrisome. Varroa destructor is considered one of the most important causes of bee colony declines. This project aims to correlate the infestation by varroa to the hemolymph sugar concentrations and bacterial and viral coinfections. Six highly infested and six control hives were compared over time. Pooled hemolymph samples from honeybees were collected for sugar concentration measurements using a previously validated portable glucometer. The hemolymph samples were submitted for bacteriology. Multiplex RT-PCR analysis was performed on honeybees for six viruses: DWV-A, DWV-B, BQCV, ABPV, KBV, and IAPV. There was also no predominance of pathogenic bacteria. In September, sugar concentrations in hemolymph were significantly lower in highly infested hives than in control hives. Infested hives showed markedly higher viral loads except for ABPV. DWV-A and BQCV viral loads from highly infested hives were significantly higher in September compared to July. A continued and severe exposure to varroa leads to increased viral charges and decreased sugar concentrations, suggesting alterations in immunity, metabolism, and reserve mobilization. These parameters contribute to the weakening and mortality of the colonies.

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