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The presence of erythrocyte ghost cells (EG) in blood smears indicates intravascular haemolysis or in-vitro haemolysis. However, observer reliability in detection of EG has not been documented. Immediate blood smear preparation is advised but may not always be practical. This study investigated the reliability of EG detection and the impact of delayed blood smear preparation. Forty blood smears from dogs and cats were evaluated twice by four observers. EG development was studied in 59 dogs and nine cats, with smears prepared at 0, 2, and 24 h post-collection. Intra- and inter-observer reliability, as assessed using Krippendorff's Alpha, ranged from 0.567 and 0.949, and 0.522 and 0.848, respectively, indicating moderate to substantial agreement. No significant increase in EG occurrence was noted between 0 and 2 h, but a significant increase was observed at 24 h. The findings suggest that EG can be reliably detected and that blood smears for the evaluation of EG can be prepared up to 2 h after blood collection without compromising detection of EG., Competing Interests: Declaration of interest The authors have indicated that they have no affiliations or financial involvement with any organisation or entity with a financial interest in, or in financial competition with, the subject matter or materials discussed in this article., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Measuring trace element concentrations in tissue can be a valuable approach to monitor animal health status. Temporal variation in the absorption, transport, and storage of elements between different tissues can, however, complicate the assessment of element-health relationships. Here, we measured concentrations of selected essential (copper (Cu), zinc (Zn), selenium (Se)) and non-essential (arsenic (As), cadmium (Cd), lead (Pb)) trace elements within blood, liver, kidney, and hair of fallow deer (Dama dama; N=20) and red deer (Cervus elaphus; N=21). Using multivariate regression and structural equation models, we estimated direct and indirect linkages between tissue-specific trace element profiles and long- (body condition) and short-term (serum protein biomarkers for acute inflammation, infection, and malnutrition) health indicators. Trace element concentrations varied markedly and were weakly correlated among tissues, with the exception of Se. After accounting for sex- and site-differences in trace element concentrations, body condition of red deer was directly, and positively, associated to trace element status in liver and hair, but not in kidney. For both deer species, trace element status in blood was directly linked to serum protein status with an indirect positive association to deer body condition. For fallow deer, no direct association between trace element status and body condition was detected in any of the tissues, possibly because of elemental homeostasis, and because all individuals were in good clinical health. This study shows that hair can serve as an effective, non-invasive, biomarker in deer health assessments, yet, to fully uncover trace element-health relationships a variety of sample matrices is preferred., (© 2023. The Author(s).)

Parasites can exert a substantial influence on the ecology of wildlife populations by altering host condition. Our objectives were to estimate single and multiparasite-condition relationships for fallow deer ( Dama dama ) and red deer ( Cervus elaphus ) in Denmark and to assess potential health effects along the parasite burden gradient. Fallow deer hosted on average two endoparasite taxa per individual (min = 0, max = 5) while red deer carried on average five parasite taxa per individual (min = 2, max = 9). Body condition of both deer species was negatively related to presence of Trichuris ssp. eggs while body condition of red deer was positively related to antibodies of the protozoan Toxoplasma gondii . For the remaining parasite taxa (n = 12), we either found weak or no apparent association between infection and deer body condition or low prevalence levels restricted formal testing. Importantly, we detected a strong negative relationship between body condition and the sum of endoparasite taxa carried by individual hosts, a pattern that was evident in both deer species. We did not detect systemic inflammatory reactions, yet serology revealed reduced total protein and iron concentrations with increased parasite load in both deer species, likely due to maldigestion of forage or malabsorption of nutrients. Despite moderate sample sizes, our study highlights the importance of considering multiparasitism when assessing body condition impacts in deer populations. Moreover, we show how serum chemistry assays are a valuable diagnostic tool to detect subtle and sub-clinical health impacts of parasitism, even at low-level infestation., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Thromboelastography (TEG) provides a global assessment of hemostasis and fibrinolysis and has broad applications to identify and monitor coagulation dysfunction in veterinary patients. Although alpacas are susceptible to a wide variety of coagulopathies, the assessment of TEG has not been reported in clinically healthy alpacas to date. The purpose of this study was to evaluate the analytical performance of recombinant human tissue factor (rhTF)- and kaolin-activated TEG and to establish reference intervals for TEG parameters (reaction [R] and clotting [K] times, angle [α], maximum amplitude [MA], and shear elastic modulus [G]) in healthy, adult alpacas. Kaolin and rhTF-activated TEG were performed using citrated whole blood samples from 20 clinically healthy, nonpregnant, adult Huacaya alpacas each after 30 min of sample storage at room temperature. Six individuals of a related species, dromedary camels, were also sampled for comparative purposes. All data were presented descriptively, assessed for normality, and compared using either independent-sample t tests or Mann-Whitney U tests, with P ≤ 0.05 considered significant. Reference intervals were calculated using a robust method and Box-Cox-transformed data. Mean TEG values (reference intervals) were determined for rhTF-activated TEG as follows: R 6.99 min (3.41-12.71), K 3.43 min (1.61-6.42), α 48.51° (27.21-67.38), MA 52.05 mm (21.53-65.92), and G 5.71 kdyn/cm 2 (1.87-9.60), while mean values (reference intervals) for kaolin-activated TEG included R 7.72 min (4.48-11.43), K 4.24 min (2.03-9.20), α 45.06° (23.66-64.20), MA 52.18 mm (33.49-66.63), and G 5.78 kdyn/cm 2 (NR-9.66). None of the measured TEG values differed significantly between activators, suggesting that activator choice may have a limited effect on TEG parameters in healthy alpacas. TEG results in alpacas were comparable to those of dromedary camels. These results will thus provide a useful starting point in the evaluation of hemostasis in adult camelids.

Objective disease markers in the southern white rhinoceros ( Ceratotherium simum simum ) are in high demand. In the field, such markers are typically needed to decide whether a captured white rhinoceros is fit to cope with quarantine, transport, or both. Captive white rhinoceros have a need for unbiased biomarkers for early detection of disease. Acute phase proteins, including haptoglobin, are proteins that significantly change their plasma concentration in response to tissue perturbation or inflammation, such as that occurring during infection or neoplastic disease. Acute phase proteins are well known diagnostic tools in both human and veterinary medicine. In this study, an ELISA with commercially available anti-human haptoglobin antibodies for quantification of haptoglobin in white rhinoceros serum was developed. The validity of the haptoglobin assay and haptoglobin as a biomarker of disease was investigated with the use of serum samples from both captive and free-ranging animals with a well-described health status. The assay was precise (intra-assay and interassay reproducibility were 5.0% and 13.1%, respectively) and reliably quantified white rhinoceros haptoglobin serum concentrations consuming low volumes of sample. The assay was sensitive to the presence of free hemoglobin in the sample at levels corresponding to a visibly hemolyzed sample. Haptoglobin was readily measurable, baseline levels (in white rhinoceros with no clinical signs of disease) did not differ between genders, and a significant increase was seen in captive as well as in free-ranging white rhinoceros with inflammatory disease. Thus, haptoglobin is a positive acute phase protein in southern white rhinoceros with potential for use as an objective marker of disease.

Acute-phase reactants indicate inflammation and are increasingly used in veterinary medicine to indicate and to monitor progression of disease. Hemostasis and inflammation have interconnected pathophysiologic pathways and influence each other on different levels. This study established observed normal ranges for acute-phase reactants and for coagulation and thromboelastographic (TEG) parameters in 49 dromedary camels ( Camelus dromedarius) and assessed the response to chronic and acute inflammation. Chronically infected animals suffering from lymph abscessation due to Corynebacterium spp. had significantly higher concentrations of the acute-phase reactants haptoglobin ( P < 0.005) and fibrinogen ( P < 0.013) and an increased clot strength characterized by an increase of the TEG parameters MA ( P < 0.039), representing the maximum amplitude of the clot strengths, and G, the global clot strength ( P < 0.022), compared to healthy animals. When the acute-phase and hemostatic responses of 10 males receiving a gonadotropin-releasing hormone vaccine and of 9 males that were surgically castrated over 7 days were studied, haptoglobin proved to be a minor positive acute-phase protein, with moderate levels in healthy animals. It increased significantly after both vaccination and castration and remained elevated 7 days postinsult. The negative reactant iron significantly decreased over the 7-day period after castration, whereas a similar decrease following vaccination lasted less than 3 days. Fibrinogen reacted as a positive, minor reactant, with a significant increase and a peak on days 3-5, with higher values seen after castration. Prothrombin time showed a slight shortening at days 5-7, and the TEG parameters MA and G showed significantly increased values, similar to fibrinogen. The acute-phase protein serum amyloid A showed poor repeatability, suggesting that the assay was not reliable.