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Background: There are few reports in dogs that have evaluated the utility of semi-quantitative scoring of bone marrow iron stores in conjunction with reticulocyte hemoglobin (CHr) to identify iron-restricted erythropoiesis due to absolute iron deficiency or iron sequestration., Objectives: An established system for scoring iron stores in human bone marrow samples was applied to dogs. The objectives were to evaluate interobserver agreement (Κ ω ), determine marrow iron scores in dogs without detectable hematologic abnormalities, and assess combined interpretation of iron scores and CHr to evaluate for iron-restricted erythropoiesis., Methods: Four blinded observers independently scored iron in 139 Prussian blue-stained canine marrow samples from 0 (none) to 6 (very heavy), including healthy controls (n = 12), clinically ill dogs with (n = 100) and without (n = 16) detectable hematologic abnormalities, and dogs with experimental nutritional iron deficiency (n = 11). Additional medical record data were available for 118 dogs to evaluate for other evidence of iron deficiency (abnormal CHr, RBC indices, serum iron variables, external blood loss, or nutritional deficiencies)., Results: Mean Κ ω was 0.69 (substantial agreement) for all samples but was 0.44 (moderate agreement) for samples with iron scores <3, indicating distinguishing scores 0-2 may not be reliable. Dogs without detectable hematologic abnormalities had scores from 3-5. Dogs with scores <3 and decreased CHr often had more indicators of iron deficiency vs dogs only having low iron scores or low CHr., Conclusions: Evaluation of dogs with marrow iron score <3 for external blood loss or nutritional deficiencies is likely clinically worthwhile, particularly if there is also decreased CHr., (© 2023 American Society for Veterinary Clinical Pathology.)

Given the move toward competency-based veterinary education and the subsequent reevaluation of veterinary curricula, there is a need for specialties to provide guidance to veterinary college administrators and educators on the core knowledge and skills pertaining to their specialty to ensure their inclusion in revised or redesigned curricula. The American Society for Veterinary Clinical Pathology (ASVCP) Education Committee sought to create a list of competencies specific to clinical pathology expected of graduating veterinarians. The stimulus for this project was the American Veterinary Medical Association Council on Education Standards of Accreditation for Colleges of Veterinary Medicine, further driven by the 2018 publication of the Association of American Veterinary Medical Colleges Competency-Based Veterinary Education Working Group framework. The recommendations made in this document are the culmination of the 2016 ASVCP Education Forum for Discussion, multiple remote subcommittee communications, and feedback obtained from ASVCP membership. The final framework includes 8 clinical pathology-focused domains of competence with 20 clinical pathology competencies and 61 clinical pathology illustrative sub-competencies. The clinical pathology-focused domains of competence are: the pre-analytical phase of testing, laboratory medicine and instrumentation, principles of test selection and interpretation, hematology and hemostasis, chemistry, endocrinology, urinalysis, and cytology. These are not intended to replace the nine established AAVMC domains of competence with supportive competencies and illustrative sub-competencies but to guide institutions for how clinical pathology aligns within the competency-based veterinary education (CBVE) framework for the practice-ready veterinary graduate. This clinical pathology competency framework may prove useful and empowering during discussions of curriculum revisions and redesigns.

This study aimed to determine the pharmacokinetics of prednisolone following intravenous and oral administration in healthy adult alpacas. Healthy adult alpacas were given prednisolone (IV, n = 4), as well as orally (PO, n = 6). Prednisolone was administered IV once (1 mg/kg). Oral administration was once daily for 5 days (2 mg/kg). Each treatment was separated by a minimum 4 month washout period. Samples were collected at 0 (pre-administration), 0.083, 0.167, 0.25, 0.5, 0.75, 1, 2, 4, 8, 12, and 24 h after IV administration, and at 0 (pre-administration), 0.25, 0.5, 0.75, 1, 2, 4, 8, 12, 24 after the first and 5 th PO administration. Samples were also taken for serial complete blood count and biochemistry analysis. Prednisolone concentration was determined by high pressure liquid chromatography. Non-compartmental pharmacokinetic parameters were then determined. After IV administration clearance was 347 mL/kg/hr, elimination half-life was 2.98 h, and area under the curve was 2,940 h * ng/mL. After initial and fifth oral administration elimination half-life was 5.27 and 5.39 h; maximum concentration was 74 and 68 ng/mL; time to maximum concentration was 2.67 and 2.33 h; and area under the curve was 713 and 660 hr * ng/mL. Oral bioavailability was determined to be 13.7%. Packed cell volume, hemoglobin, and red blood cell counts were significantly decreased 5 days after the first PO administration, and serum glucose was significantly elevated 5 days after the first PO administration. In conclusion, serum concentrations of prednisolone after IV and PO administration appear to be similar to other veterinary species. Future research will be needed to determine the pharmacodynamics of prednisolone in alpacas., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Videla, Sommardahl, Smith, Schaefer and Cox.)

Background: There are few effective drugs for treatment of seizures in avian species., Objectives: To investigate the pharmacokinetics and safety of zonisamide in chickens., Methods: Phase 1: chickens (n = 4) received a single oral dose of zonisamide at 20 mg/kg. Blood samples were collected intermittently for 36 hr after dosing. Phase 2: chickens (n = 8) received zonisamide in a dose escalation protocol (20, 30, 60 and 80 mg/kg orally every 12 hr). The dose was increased weekly, and peak and trough blood samples were collected on Days 1, 3, and 7 each week. Two birds were randomly euthanized at the end of each week. Plasma zonisamide concentrations were analysed using a commercial immunoassay. Drug concentration vs. time data were subjected to non-compartmental pharmacokinetic analysis., Results: For Phase 1, peak plasma zonisamide (C max ) was 15 ± 3 µg/ml at 2 ± 1 hr (T max ). The disappearance half-life was 6.5 ± 1 hr. Mean plasma concentrations remained within the (human) therapeutic range (10-40 µg/ml) for 6 hr. For Phase 2 of the study, plasma concentrations of zonisamide remained within or close to the recommended mammalian therapeutic range for birds in the 20 and 30 mg/kg dose. Area under the curve (AUC) and C max were dose dependent. Two birds developed immune-mediated haemolytic anaemia., Conclusions: Zonisamide appears to be a viable drug for use in chickens at a dose of 20 mg/kg orally every 12 hr., (© 2021 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

Background: Insufficient iron for erythropoiesis can occur in multiple conditions, including absolute iron deficiency, which is often caused by chronic external hemorrhage in dogs. Distinguishing this from other causes of iron-restricted erythropoiesis allows appropriate intervention. Decreased marrow iron assessed by Prussian blue staining is a method to diagnose absolute iron deficiency, but scoring systems for marrow iron are not validated in dogs., Objectives: Our objectives were to (a) evaluate the technical performance of two bone marrow iron scoring systems used in human medicine and (b) examine the effects of destaining and restaining on iron stores after Wright-stained marrow slides are destained and restained with a Prussian blue stain., Methods: Two marrow aspirate slides were included from each of 12 ill dogs in which marrow was collected during clinical evaluation. One slide was directly stained with Prussian blue. The other was first stained with Wright stain, then destained and restained with Prussian blue. Three blinded observers scored the presence of iron in each of the 24 randomized slides using the Gale (scale 0-6) and sideroblast methods (percentage score). Slides were then re-randomized and rescored., Results: For the Gale method, interobserver agreement was fair, and intraobserver agreement was substantial to perfect. There was less agreement using the sideroblast method, with a significant observer effect. Iron scores were significantly lower in destained slides compared with those stained directly., Conclusions: Interobserver and intraobserver agreements were acceptable for the Gale method, but the sideroblast method should be used cautiously. A destaining procedure before Prussian blue staining could decrease marrow iron scores., (© 2021 American Society for Veterinary Clinical Pathology.)

Ferret systemic coronaviral disease (FSCD) is a well-established cause of mortality in domestic ferrets. We describe herein novel findings in a case of FSCD that was diagnosed and medically managed following virus detection by immunohistochemical (IHC) staining of surgical biopsy samples. Hematologic changes in this ferret suggested spread of the virus to the bone marrow, which was confirmed by IHC staining of a postmortem sample. Genotyping of the virus indicated that the virus grouped with alphacoronaviruses and was most closely related to ferret enteric coronavirus (FRECV) MSU-2. Our clinical case demonstrates that a FRECV MSU-2-like ferret coronavirus associated previously with the enteric pathotype may cause systemic disease, including bone marrow involvement causing persistent pancytopenia.

Defects in the Fanconi anemia (FA) DNA damage-response pathway result in genomic instability, developmental defects, hematopoietic failure, cancer predisposition, and metabolic disorders. The endogenous sources of damage contributing to FA phenotypes and the links between FA and metabolic disease remain poorly understood. Here, using mice lacking the Fancd2 gene, encoding a central FA pathway component, we investigated whether the FA pathway protects against metabolic challenges. Fancd2 -/- and wildtype (WT) mice were fed a standard diet (SD), a diet enriched in fat, cholesterol, and cholic acid (Paigen diet), or a diet enriched in lipid alone (high-fat diet (HFD)). Fancd2 -/- mice developed hepatobiliary disease and exhibited decreased survival when fed a Paigen diet but not a HFD. Male Paigen diet-fed mice lacking Fancd2 had significant biliary hyperplasia, increased serum bile acid concentration, and increased hepatic pathology. In contrast, female mice were similarly impacted by Paigen diet feeding regardless of Fancd2 status. Upon Paigen diet challenge, male Fancd2 -/- mice had altered expression of genes encoding hepatic bile acid transporters and cholesterol and fatty acid metabolism proteins, including Scp2/x , Abcg5/8 , Abca1 , Ldlr , Srebf1 , and Scd-1 Untargeted lipidomic profiling in liver tissue revealed 132 lipid species, including sphingolipids, glycerophospholipids, and glycerolipids, that differed significantly in abundance depending on Fancd2 status in male mice. We conclude that the FA pathway has sex-specific impacts on hepatic lipid and bile acid metabolism, findings that expand the known functions of the FA pathway and may provide mechanistic insight into the metabolic disease predisposition in individuals with FA., (© 2019 Moore et al.)

Background: Mycoplasma haemolamae (Mhl) and gastrointestinal nematodes can cause anemia in camelids. Control programs aim to suppress parasitism without promoting anthelminthic resistance, but few evidence-based guidelines define acceptable parasite loads in camelids., Hypothesis/objectives: In clinically healthy nonanemic camelids, compare erythrocyte variables to Mhl real-time PCR status and to fecal egg count (FEC). Determine the FEC threshold above which erythrocyte variables are consistently below reference interval medians., Animals: One hundred fourteen client-owned adult alpacas and llamas., Methods: In a cross-sectional study, whole blood in ethylenediaminetetraacetic acid (EDTA) was assessed for packed cell volume (PCV) by centrifugation, erythrocyte count (RBC), and hemoglobin concentration (HGB) using an ADVIA120 analyzer, and Mhl using real-time PCR. Trichostrongyle eggs per gram (epg) were counted by modified McMaster test on freshly collected feces. Significant differences in erythrocyte variables based on Mhl status and FEC thresholds were assessed by independent t test and one-way ANOVA, respectively., Results: Packed cell volume, RBC, and HGB were not significantly different between Mhl-positive and Mhl-negative animals, but were significantly lower in animals with FEC >1000 epg compared to those with <500 epg. All animals with FEC >600 epg had RBC and HGB below the reference interval median. All animals with FEC >750 epg had PCV below the reference interval median., Conclusions and Clinical Importance: In healthy nonanemic camelids, positive Mhl PCR is not associated with lower erythrocyte variables and such animals may not warrant treatment. Fecal egg count >600-750 epg has a negative effect on erythrocyte variables, and may be a guide for deworming protocols., (© 2019 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

Background: Accurate erythrocyte measurements with ADVIA hematology analyzers require isovolumetric cell sphering in one reaction and hemolysis in another. However, camelid erythrocytes are resistant to sphering and osmotic lysis, and no published evaluation of ADVIA methods for camelids exists., Objectives: The objectives were to demonstrate whether camelid erythrocytes sphere in the ADVIA red blood cell/platelet (RBC/PLT) reagent and lyse in the ADVIA cyanide HGB reagent, and to determine optimal ADVIA settings for camelids., Methods: Camelid and canine blood were diluted to 1:625 in RBC/PLT reagent and evaluated microscopically for erythrocyte sphering. A camelid sample was incubated with the hemoglobin (HGB) reagent at varying dilutions to evaluate hemolysis. The RBC, hematocrit (HCT), mean cell volume (MCV), and mean corpuscular hemoglobin concentration (MCHC) using three ADVIA species settings (equine, bovine, and caprine) were compared to their respective reference methods: Z2 Coulter impedance counter, packed cell volume, calculated MCV (PCV × 10/Coulter RBC), and calculated MCHC (HGB × 100/PCV). Reference MCV was also compared to MCV calculated using the ADVIA equine RBC count. Comparisons were assessed using Passing-Bablok regression and Bland-Altman difference plots., Results: Camelid erythrocytes did not sphere in the RBC/PLT reagent, but did lyse in the HGB reagent. The ADVIA equine setting RBC count was acceptably close to the Coulter count. Hematocrit, MCV, and MCHC from all settings were significantly different from the reference methods. Mean cell volumes calculated using the equine setting RBC counts were acceptably close to the reference MCV., Conclusions: Camelid ADVIA erythrogram results should be reported as follows: RBC counts and HGB concentrations using the equine setting, spun PCVs, MCVs calculated using the PCV and equine setting RBC, and MCHCs calculated using the PCV and equine setting HGB., (© 2019 American Society for Veterinary Clinical Pathology.)

OBJECTIVE To provide contemporary preliminary guidelines for the morphological evaluation of bone marrow in conjunction with CBC results for healthy juvenile (3- to 6-month-old) female New Zealand White rabbits (Oryctolagus cuniculus). ANIMALS 22 female New Zealand White rabbits. PROCEDURES Each rabbit was sedated, and a blood sample (3 mL) was collected from an ear artery for a CBC, after which the rabbit was euthanized. Within 5 minutes after euthanasia, bone marrow samples were obtained from the femur for cytologic and histologic evaluation. Bone marrow specimens for cytologic evaluation were stained with modified Wright stain, and those for histologic evaluation were stained with either H&E or Prussian blue stain. RESULTS The CBC results were within published reference ranges for all rabbits except 4, each of which had mild leukopenia. Cytologic assessment of bone marrow revealed a median myeloid-to-erythroid ratio of 0.7 and 2.8 megakaryocytes/low-power field (magnification, 100X), and the median percentages of lymphocytes, plasma cells, and macrophages were 11.5%, 0.1%, and 0%, respectively. The myeloid-to-erythroid ratio was not significantly correlated with any CBC variable. On histologic evaluation of bone marrow, the cellularity ranged from 30% to 50%, there were 2.1 to 7.7 megakaryocytes/hpf (magnification, 400X), and no iron stores were visible in H&E or Prussian blue-stained specimens. CONCLUSIONS AND CLINICAL RELEVANCE Results of the present study provided contemporary preliminary guidelines for the evaluation of bone marrow in healthy laboratory rabbits.