Covariation between glucocorticoids, behaviour and immunity supports the pace-of-life syndrome hypothesis: an experimental approach.

The biomedical literature has consistently highlighted that long-term elevation of glucocorticoids might impair immune functions. However, patterns are less clear in wild animals. Here, we re-explored the stress-immunity relationship considering the potential effects of behavioural profiles. Thirteen captive roe deer (Capreolus capreolus) were monitored over an eight-week period encompassing two capture events. We assessed how changes in baseline faecal cortisol metabolite (FCM) concentrations following a standardized capture protocol and an immune challenge using anti-rabies vaccination affected changes in 13 immune parameters of innate and adaptive immunity, and whether these changes in baseline FCM levels and immune parameters related to behavioural profiles. We found that individuals with increased baseline FCM levels also exhibited increased immunity and were characterized by more reactive behavioural profiles (low activity levels, docility to manipulation and neophilia). Our results suggest that the immunity of large mammals may be influenced by glucocorticoids, but also behavioural profiles, as it is predicted by the pace-of-life syndrome hypothesis. Our results highlight the need to consider covariations between behaviour, immunity and glucocorticoids in order to improve our understanding of the among-individual variability in the stress-immunity relationships observed in wildlife, as they may be underpinned by different life-history strategies.

Detection of circulating microfilariae in canine EDTA blood using lens-free technology: preliminary results.

Dirofilaria immitis causes life-threatening heart disease in dogs, thus screening of dog populations is important. Lens-free technology (LFT) is a low-cost imaging technique based on light diffraction that allows computerized recognition of small objects in holographic images. We evaluated an algorithm capable of recognizing microfilariae in canine whole blood using the LFT. We examined 3 groups of 10 EDTA blood specimens, from dogs with microfilaremia (group A), healthy dogs (B), and dogs with hematologic modifications other than microfilaremia (C). The LFT analyzer photographed repeated series of 5 images of all samples. The algorithm declared a sample positive if a microfilaria was detected on ≥1, ≥2, or ≥3 of the 5 images of a series. Microfilariae were detected visually in the images in 9 of 10 cases in group A; no microfilariae were seen in the images from groups B and C. Of the 30 cases, there were 14, 4, and only 3 false-positives with the 1 of 5, 2 of 5, and 3 of 5 image cutoffs, respectively. There were no false-negatives, regardless of cutoff. LFT seems useful for detecting microfilaria and could have application in clinical pathology.

Abnormal Sysmex XT-2000iV DIFF scattergram in a cat with a prominent mastocytemia.

A 2-year-old neutered male domestic shorthair cat was presented to the emergency service of the National Veterinary School of Toulouse (France) for acute vomiting and diarrhea with lethargy, inappetence, and adypsia for the past 48 hours. Complete blood counts were performed with the ProCyte DX at the emergency department and with the Sysmex XT-2000iV at the laboratory 2 weeks later. The scattergrams from the two analyzers revealed similar unusual and abnormal dot plots. The Sysmex XT-2000iV DIFF scattergram also showed no clear separation between different leukocyte populations. The eosinophil cluster was in an abnormal location compared with that of the "typical" location in a normal cat. A blood smear evaluation revealed the presence of numerous mast cells. Thus, we hypothesized that the Sysmex XT-2000iV had detected the mast cell population, and this led to errors in the differential counts. To explore this hypothesis, we manually gated on the DIFF scattergram and performed a manual differential on the blood smear. With this new gating strategy, the Sysmex XT-2000iV and manual differentials were similar. Thus, in the case of systemic mastocytosis, mast cells can be located between the lymphocyte, monocyte, and eosinophil clusters on scattergrams.

The effects of storage at 4°C and 20°C on the hemograms of C57BL/6 mice and Wistar rats using the IDEXX ProCyte Dx and blood smear evaluations.

BACKGROUND: Delayed blood analysis might be unavoidable in laboratory practice, but little is known about rodent blood stability, especially cell morphology and scattergram results. OBJECTIVES: This study aimed to evaluate the stability of rodent blood cell counts and morphologies at different temperatures using the ProCyte Dx analyzer and performing manual observations. METHODS: Ten Wistar rats and 10 C57bl/6 mice were sampled on EDTA tubes and aliquoted for storage (4°C, 20°C). Hematologic analyses were performed immediately and at T6h, T24h, T48h (rats and mice), and T72h (rats only) after storage. RESULTS: In rats, at any temperature, red blood cell counts, hemoglobin concentrations (HGB), mean corpuscular hemoglobin (MCH) levels, and reticulocyte, white blood cell (WBC), eosinophil, and impedance platelet counts remained stable over time. The main changes were observed at 20°C for hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and WBC differential counts. Optical platelet counts (PLT-O) and platelet variables underwent changes at both temperatures from T24h. In mice, red blood cell counts by impedance (RBC-I), MCH, and WBC, lymphocyte, eosinophil, and platelet counts, and plateletcrit (PCT) were stable over time and at all temperatures. As in rats, the most significant changes were observed at 20°C and concerned the optical RBC (RBC-O) counts, HCTs, MCVs, MCHCs, and reticulocyte, neutrophil, and monocyte counts. For both species, blood cell morphologies were altered from T24h at all temperatures, and platelet clumps were more numerous at 4°C. CONCLUSIONS: When rodent blood analyses need to be delayed, storage at 4°C is preferred and should not exceed 24 hours. PLT counts should be interpreted cautiously in refrigerated specimens with mandatory blood smear evaluations when abnormal scattergrams are observed.