Dehorning Does Not Alter the Stress Response in Southern White Rhinoceroses (Ceratotherium simum simum) during Transport: A Preliminary Investigation.

Translocation and dehorning are common and important practices for rhinoceros management and conservation. It is not known if dehorning causes a stress response or negatively affects rhinoceroses during transport. Twenty-three subadult wild Southern white rhinoceros (Ceratotherium simum simum) bulls were immobilized and translocated >280 km for population management reasons. Ten animals were dehorned at capture, and 13 animals were transported without dehorning. For transport, five dehorned and six nondehorned rhinoceroses were sedated with azaperone (62.38±9.54 µg/kg) and five dehorned and seven nondehorned rhinoceroses with midazolam (64.61±9.28 µg/kg). Blood samples were collected at capture, start of transport, and after 6 h of transport. Measurements included 10 physiologic variables: hematocrit, total serum protein, creatine kinase (CK), aspartate aminotransferase, gamma-glutamyl transferase (GGT), creatinine, urea, cholesterol, ß-hydroxybutyrate, and glucose; and four stress response variables: cortisol, epinephrine, neutrophil-to-lymphocyte ratio, and leukocyte coping capacity. Using a linear mixed model, CK and GGT were higher in dehorned compared with nondehorned rhinoceroses. There were no significant differences in the other variables between the two groups. The likely cause of these differences is that dehorned animals spent more time in the crate before the start of transport than nondehorned rhinoceroses (3:11±0:54 h vs. 1:12±0:56 h, P<0.001). These results indicate that dehorning does not negatively alter the white rhinoceros' physiologic and stress responses during translocation, supporting its use for antipoaching measures.

Guidelines for resident training in veterinary clinical pathology. IV: Laboratory quality management-Teaching domains, competencies, and suggested learning outcomes.

BACKGROUND: The 2019 ASVCP Education Committee Forum for Discussion, presented at the annual ASVCP/ACVP meeting, identified a need to develop recommendations for teaching laboratory quality management principles in veterinary clinical pathology residency training programs. OBJECTIVES: To present a competency-based framework for teaching laboratory quality management principles in veterinary clinical pathology residency training programs, including entrustable professional activities (EPAs), domains of competence, individual competencies, and learning outcomes. METHODS: A joint subcommittee of the ASVCP Quality Assurance and Laboratory Standards (QALS) and Education Committees executed this project. A draft guideline version was reviewed by the ASVCP membership and shared with selected ACVP committees in early 2022, and a final version was voted upon by the full QALS and Education Committees in late 2022. RESULTS: Eleven domains of competence with relevant individual competencies were identified. In addition, suggested learning outcomes and resource lists were developed. Domains and individual competencies were mapped to six EPAs. CONCLUSIONS: This guideline presents a framework for teaching principles of laboratory quality management in veterinary clinical pathology residency training programs and was designed to be comprehensive yet practical. Guidance on pedagogical terms and possible routes of implementation are included. Recommendations herein aim to improve and support resident training but may require gradual implementation, as programs phase in necessary expertise and resources. Future directions include the development of learning milestones and assessments and consideration of how recommendations intersect with the American College of Veterinary Pathologists training program accreditation and certifying examination.

Regenerative response in dogs naturally and experimentally infected with Babesia rossi.

BACKGROUND: The regenerative response following Babesia rossi infection in dogs is mild, despite severe hemolytic anemia. OBJECTIVE: We aimed to compare the admission absolute reticulocyte count (ARC) and reticulocyte indices in 103 dogs naturally infected with B. rossi with 10 dogs suffering from immune-mediated hemolytic anemia (IMHA) and 14 healthy control dogs. The regenerative response was also evaluated in five dogs experimentally infected with B. rossi. METHODS: This is a retrospective observational study of records generated on the ADVIA 2120 hematology analyzer. RESULTS: The median hematocrits (HCT) of the B. rossi and IMHA groups were significantly lower than the control group (p < .001 for both); however, no differences were seen between the B. rossi and IMHA groups. Compared with the control group, the median ARC was significantly higher in the B. rossi (p = .006) and IMHA (p = .019) groups but significantly lower in the B. rossi group than the IMHA group (p = .041). In the experimentally infected dogs, there was a sudden decrease in the ARC approximately 48 h after the detection of peripheral parasitemia, which was followed by an increase after treatment. Reticulocytes of naturally infected B. rossi dogs were larger, with more variation in cellular volume. The reticulocytes of the experimentally infected dogs decreased in size with decreasing hemoglobin concentrations as the study progressed. CONCLUSIONS: The regenerative response in dogs naturally infected with B. rossi is inadequate, given the severity of the anemia observed, and it might be a result of direct suppressive action by the parasite or host response on the bone marrow.

Acute phase reactants in nondomesticated mammals-A veterinary clinical pathology perspective.

Applications for acute phase reactants (APRs) in nondomesticated mammals include identifying inflammatory disease, monitoring the course of specific disease processes and recovery during rehabilitation, detecting preclinical or subclinical disease, being used as bioindicators for monitoring population and ecosystem health, and as markers of stress and animal welfare. Serum amyloid A, haptoglobin, C-reactive protein, fibrinogen, albumin, and iron are most commonly measured. The procedure for evaluating an APR in a nondomesticated mammalian species should follow a stepwise approach beginning with an assessment of analytical performance, followed by an evaluation of overlap performance, clinical performance, and impact on patient outcomes and management. The lack of species-specific standards and antibodies for nondomesticated mammals presents a challenge, and more attention needs to be focused on assessing cross-reactivity and ensuring adequate analytical performance of APR assays. Sample selection for the initial evaluation of APRs should consider preanalytical influences and should originate from animals with confirmed inflammatory disease and healthy animals. Reference intervals should be generated according to published guidelines. Further evaluation should focus on assessing the diagnostic utility of APRs in specific disease scenarios relevant to a species. Greater attention should be paid to assay performance and uniformity of methods when using APRs for population and ecosystem surveillance. Veterinary clinical pathologists should work closely with zoo veterinarians and wildlife researchers to optimize the accuracy and utility of APR measurements in these various conservation medicine scenarios.

Quality Assurance for Veterinary In-Clinic Laboratories.

Quality assurance and the implementation of a quality management system are as important for veterinary in-clinic laboratories as for reference laboratories. Elements of a quality management system include the formulation of a quality plan, establishment of quality goals, a health and safety policy, trained personnel, appropriate and well-maintained facilities and equipment, standard operating procedures, and participation in external quality assurance programs. Quality assurance principles should be applied to preanaltyic, analytic, and postanalytic phases of the in-clinic laboratory cycle to ensure that results are accurate and reliable and are released in a timely manner.

Markers of inflammation in free-living African elephants (Loxodonta africana): Reference intervals and diagnostic performance of acute phase reactants.

INTRODUCTION: Acute phase reactants (APRs) have not been investigated in free-living African elephants (Loxodonta africana), and there is little information about negative APRs albumin and serum iron in elephants. OBJECTIVES: We aimed to generate reference intervals (RIs) for APRs for free-living African elephants, and to determine the diagnostic performance of APRs in apparently healthy elephants and elephants with inflammatory lesions. METHODS: Stored serum samples from 49 apparently healthy and 16 injured free-living elephants were used. The following APRs and methods were included: albumin, bromocresol green; haptoglobin, colorimetric assay; serum amyloid A (SAA), multispecies immunoturbidometric assay, and serum iron with ferrozine method. Reference intervals were generated using the nonparametric method. Indices of diagnostic accuracy were determined by receiver-operator characteristic (ROC) curve analysis. RESULTS: Reference intervals were: albumin 41-55 g/L, haptoglobin 0.16-3.51 g/L, SAA < 10 mg/L, and serum iron 8.60-16.99 µmol/L. Serum iron and albumin concentrations were lower and haptoglobin and SAA concentrations were higher in the injured group. Serum iron had the best ability to predict health or inflammation, followed by haptoglobin, SAA, and albumin, with the area under the ROC curve ranging from 0.88-0.93. CONCLUSIONS: SAA concentrations were lower in healthy African vs Asian elephants, and species-specific RIs should be used. Serum iron was determined to be a diagnostically useful negative APR which should be added to APR panels for elephants.

Experimental infection with African Horse Sickness Virus in horses induces only mild temporal hematologic changes and acute phase reactant response.

OBJECTIVE: African Horse Sickness (AHS) is a vector-borne disease endemic to sub-Saharan Africa caused by African Horse Sickness Virus (AHVS). Infections in naïve horses have high morbidity and mortality rates. AHS pathogenesis is not well understood; neither the hematologic changes nor acute phase response occurring during infection has been fully evaluated. The study's objective was to characterize the hematologic changes and acute phase response during experimental infection with AHSV. ANIMALS: 4 horses negative for AHSV group-specific antibodies. PROCEDURES: In this prospective, longitudinal study conducted between November 23 and December 2, 2020, horses were experimentally infected with AHSV, and blood samples were obtained before inoculation and then every 12 hours until euthanasia. Hematologic changes and changes for serum amyloid A (SAA) and iron concentration were evaluated over time using a general linear model including natural logarithm of sampling time. RESULTS: All horses were humanely euthanized due to severe clinical signs typical of AHS. Median Hct increased significantly, and the median WBC count, monocyte count, eosinophil count, and myeloperoxidase index changed significantly in all horses over time. Horses developed marked thrombocytopenia (median, 48 X 103 cells/µL; range, 21 X 103 to 58 X 103 cells/µL) while markers of platelet activation also changed significantly. Median SAA increased and serum iron concentration decreased significantly over time. CLINICAL RELEVANCE: Results indicated severe thrombocytopenia with platelet activation occurs during infection with AHSV. Changes in acute phase reactants SAA and iron, while significant, were unexpectedly mild and might not be useful clinical markers.

ASSESSMENT OF CAPILLARY ZONE ELECTROPHORESIS AND SERUM AMYLOID A QUANTITATION IN CLINICALLY NORMAL AND ABNORMAL SOUTHERN WHITE RHINOCEROS (CERATOTHERIUM SIMUM SIMUM) AND SOUTHERN BLACK RHINOCEROS (DICEROS BICORNIS MINOR).

Capillary zone electrophoresis (CZE) and an immunoassay for serum amyloid A (SAA) were used to examine serum samples from clinically normal and abnormal southern white rhinoceros (Ceratotherium simum simum) and southern black rhinoceros (Diceros bicornis minor) under managed care. CZE resolved seven fractions as well as subfractions for α1 globulins. Reference intervals were calculated for white rhinoceros (n = 33) and found to have some differences over previously reported intervals generated using agarose gel electrophoresis (AGE) methods in sera from free-ranging animals. In addition, the coefficient of variation related to fraction quantitation was found to be overlapping or superior to that reported for AGE. No significant differences were observed in CZE measurands and total protein between clinically normal and abnormal rhinoceros. In contrast to CZE, significant differences in SAA levels (P < 0.001) were observed in samples from the white rhinoceros between clinically normal and abnormal animals. In addition, in limited sample sets with repeated measures, SAA provided prognostic value. Future studies should generate more robust reference intervals and delineate the application of both SAA quantitation and CZE in routine health assessments and in prognostication.

Comparison of three hematocrit measurement methods in the southern white rhinoceros (Ceratotherium simum simum).

BACKGROUND: Hematocrit (HCT) determination is an integral part of health and disease assessments in captive and wild white rhinoceroses. Several affordable automated hematology analyzers have been developed for in-clinic and field use and have the advantage of being able to measure a large number of additional measurands. However, the accuracy of these analyzers for rhinoceros HCT measurements has not yet been investigated. OBJECTIVES: We aimed to compare the HCT results generated by the EPOC portable analyzer system and the Abaxis VetScan HM5 with the gold standard of a manual packed cell volume (PCV) measured using the microhematocrit method. METHODS: Hematocrits were measured with the EPOC and the Abaxis VetScan HM5 (bovine setting) and compared with the PCVs of 69 white rhinoceros whole blood samples. Results were compared using Bland-Altman difference plots and Passing-Bablok regression analysis. A total allowable analytical error of 10% was set as the performance goal. RESULTS: A significant positive bias, with a mean of 7.7% for the EPOC and 17.9% for the Abaxis, was found compared with the manual PCV method. CONCLUSIONS: The allowable error goal of 10% was not exceeded with the EPOC analyzer. Although not analytically equivalent to the gold standard, the EPOC results could therefore be used as approximations in critical situations where manual measurements cannot be performed. The Abaxis exceeded this allowable error and overestimated HCTs in rhinoceroses. Therefore, method-specific reference intervals should be used.

Serum cobalamin concentrations in dogs infected with canine parvoviral enteritis.

OBJECTIVE: To compare the serum cobalamin concentrations in canine parvovirus (CPV)-infected dogs with those of healthy control dogs. ANIMALS: 45 dogs with CPV enteritis and 17 healthy age-matched control dogs. PROCEDURES: Infection was confirmed by visualization of CPV-2 through fecal electron microscopy. All dogs received supportive care. Serum samples taken at admission were used to determine cobalamin, C-reactive protein, and albumin concentrations. RESULTS: Serum cobalamin concentrations were significantly lower in the CPV-infected group (median [interquartile range], 173 pmol/L [< 111 to 722 pmol/L]) than in healthy control dogs (379 pmol/L [193 to > 738 pmol/L). There was no association between cobalamin concentration and C-reactive protein or albumin concentration. CLINICAL RELEVANCE: While hypocobalaminemia was common in CPV-infected dogs, the clinical relevance of this finding remains to be determined. Studies assessing markers of cellular cobalamin deficiency in dogs with CPV infection appear warranted.

Reference Intervals for Selected Hematology and Clinical Chemistry Measurands in Temminck's Pangolin (Smutsia temminckii).

Pangolins are the world's most trafficked non-human mammals. A significant number of Temminck's pangolins (Smutsia temminckii) are presented for veterinary care and rehabilitation in southern Africa. Little is known about the physiology and normal health of this species, making diagnosis and medical management difficult. This study aimed to establish reference intervals (RIs) for hematology and plasma clinical chemistry in the Temminck's pangolin. RIs were generated according to international guidelines using samples from 27 healthy free-living (n = 18) and rehabilitated (n = 9) pangolins. Hematology was performed using the Abaxis VetScan HM5 analyzer with manual differentials; clinical chemistry was performed using heparin plasma on the Abaxis VetScan VS2 and Cobas Integra 400 Plus analyzers. Hematology RIs were: RBC 3.88-8.31 × 1012/L, HGB 73-150 g/L, HCT 26-51%, MCV 59-72 fL, MCH 15.6-21.4 pg, MCHC 257-325 g/L, RDW 14.3-19.1%, WBC 1.80-10.71 × 109/L. Vetscan VS2 clinical chemistry RIs were: albumin 27-41 g/L, ALP 26-100 U/L, ALT 25-307 U/L, amylase 267-826 U/L, bilirubin 4-10 µmol/L, calcium 2.1-2.2 mmol/L, globulin 21-55 g/L, glucose 3.8-10.0 mmol/L, phosphate 1.3-2.6 mmol/L, potassium 3.6-5.9 mmol/L, sodium 132-140 mmol/L total protein 52-84 g/L, and urea 5.3-11.4 mmol/L. RIs for creatinine were not calculated as analytical imprecision exceeded analytical performance goals. Cobas Integra clinical chemistry RIs were: albumin 22-33 g/L, ALP 20-104 U/L, ALT 17-291 U/L, amylase 466-1,533 U/L, bilirubin 1-14 µmol/L, calcium 2.0-2.4 mmol/L, creatinine <58 µmol/L, globulin 23-49 g/L, glucose 3.6-10.1 mmol/L, phosphate 1.0-2.2 mmol/L, potassium 3.1-5.8 mmol/L, sodium 137-150 mmol/L, total protein 47-72 g/L, and urea 6.0-12.5 mmol/L. There was significant bias between the two chemistry analyzers for several measurands. Differences were found for some analytes between free-living and rehabilitated animals, probably reflecting differences in nutrition and hydration. These are the first RIs generated for Temminck's pangolin. These results will allow veterinarians to better determine pangolin health status, formulate optimal treatment plans and increase patient survival rates in this endangered species.

Oxidative burst and phagocytic activity of phagocytes in canine parvoviral enteritis.

Canine parvoviral enteritis (CPE) is a severe disease characterized by systemic inflammation and immunosuppression. The function of circulating phagocytes (neutrophils and monocytes) in affected dogs has not been fully investigated. We characterized the functional capacity of canine phagocytes in CPE by determining their oxidative burst and phagocytic activities using flow cytometry. Blood was collected from 28 dogs with CPE and 11 healthy, age-matched, control dogs. Oxidative burst activity was assessed by stimulating phagocytes with opsonized Escherichia coli or phorbol 12-myristate 13-acetate (PMA) and measuring the percentage of phagocytes producing reactive oxygen species and the magnitude of this production. Phagocytosis was measured by incubating phagocytes with opsonized E. coli and measuring the percentage of phagocytes containing E. coli and the number of bacteria per cell. Complete blood counts and serum C-reactive protein (CRP) concentrations were also determined. Serum CRP concentration was negatively and positively correlated with segmented and band neutrophil concentrations, respectively. Overall, no differences in phagocyte function were found between dogs with CPE and healthy control dogs. However, infected dogs with neutropenia or circulating band neutrophils had decreased PMA-stimulated oxidative burst activity compared to healthy controls. Additionally, CPE dogs with neutropenia or circulating band neutrophils had decreased PMA- and E. coli-stimulated oxidative burst activity and decreased phagocytosis of E. coli compared to CPE dogs without neutropenia or band neutrophils. We conclude that phagocytes have decreased oxidative burst and phagocytic activity in neutropenic CPE dogs and in CPE dogs with circulating band neutrophils.

CHALLENGES TO ANIMAL WELFARE DURING TRANSPORTATION OF WILD MAMMALS: A REVIEW (1990-2020).

Wild mammal transport is an important component of conservation translocation as well as the economic wildlife trade. This article reviews the physiological responses to transport that have been measured in wild mammalian species, factors associated with these responses, and interventions that have been applied to mitigate these responses. By organizing the literature review along the "five domains model" of animal welfare, namely, the physical-functional domains (nutrition, environment, health, behavior) and the mental domain (mental state), it can be demonstrated that wild mammal transport is associated with challenges to ensuring positive animal welfare in all five domains. Transported wild mammals can experience dehydration, catabolism, fatigue, immunosuppression, behavioral changes, and stress. Factors influencing these physiological responses to transport have only been researched in a few studies encompassing species, journey length, ambient temperature, vehicle motion, stocking density, orientation, habituation, vehicle speed allowance, and road type. The administration of tranquilizers has been shown to mitigate negative physiological responses to transport. There is a need to further investigate species and situation-specific physiological responses to transport and factors associated with these responses in order to identify challenges to ensuring animal welfare and improving translocation success.

Reference Intervals for Hematology and Clinical Chemistry for the African Elephant (Loxodonta africana).

The African elephant (Loxodonta africana) is listed as vulnerable, with wild populations threatened by habitat loss and poaching. Clinical pathology is used to detect and monitor disease and injury, however existing reference interval (RI) studies for this species have been performed with outdated analytical methods, small sample sizes or using only managed animals. The aim of this study was to generate hematology and clinical chemistry RIs, using samples from the free-ranging elephant population in the Kruger National Park, South Africa. Hematology RIs were derived from EDTA whole blood samples automatically analyzed (n = 23); manual PCV measured from 48 samples; and differential cell count results (n = 51) were included. Clinical chemistry RIs were generated from the results of automated analyzers on stored serum samples (n = 50). Reference intervals were generated according to American Society for Veterinary Clinical Pathology guidelines with a strict exclusion of outliers. Hematology RIs were: PCV 34-49%, RBC 2.80-3.96 × 1012/L, HGB 116-163 g/L, MCV 112-134 fL, MCH 35.5-45.2 pg, MCHC 314-364 g/L, PLT 182-386 × 109/L, WBC 7.5-15.2 × 109/L, segmented heterophils 1.5-4.0 × 109/L, band heterophils 0.0-0.2 × 109/L, total monocytes 3.6-7.6 × 109/L (means for "regular" were 35.2%, bilobed 8.6%, round 3.9% of total leukocytes), lymphocytes 1.1-5.5 × 109/L, eosinophils 0.0-0.9 × 109/L, basophils 0.0-0.1 × 109/L. Clinical chemistry RIs were: albumin 41-55 g/L, ALP 30-122 U/L, AST 9-34 U/L, calcium 2.56-3.02 mmol/L, CK 85-322 U/L, GGT 7-16 U/L, globulin 30-59 g/L, magnesium 1.15-1.70 mmol/L, phosphorus 1.28-2.31 mmol/L, total protein 77-109 g/L, urea 1.2-4.6 mmol/L. Reference intervals were narrower than those reported in other studies. These RI will be helpful in the future management of injured or diseased elephants in national parks and zoological settings.

Effects of storage time and temperature on thromboelastographic analysis in dogs and horses.

BACKGROUND: The accessibility of thromboelastography (TEG) to general practitioners is limited by short sample storage times (30 minutes) and storage temperatures (20-23°C). OBJECTIVES: We aimed to evaluate the stability of canine and equine citrated blood samples when stored for extended periods of time, both at room temperature (RT) (20-23°C) and refrigerator temperature (FT) (2-7.5°C). METHODS: Citrated whole blood samples from healthy dogs and horses (n = 10 for each) were stored for 30 minutes (baseline) at RT before TEG analysis. Baseline values for TEG variables R, K, α, MA, LY30, and LY60 were compared with those from samples stored for 2, 8, and 22.5 h, at RT and FT. Results were compared using an ANOVA (P < .05). Total allowable analytical error (TEa ) based on biological variation data was used to evaluate stability. RESULTS: In dogs, statistically significant differences included shorter R, longer K, decreased MA, and increased LY60 at various time points and storage temperatures from 2 h onward. Only samples stored for 2 h at FT showed acceptable stability compared with TEa . In horses, statistically significant differences included shorter R and K, and decreased α, LY30, and LY60 at various time points and storage temperatures from 2 h onward. Samples were not stable at any time compared with TEa , regardless of the temperature. CONCLUSIONS: In this study, canine samples could be stored for up to 2 h at FT without affecting TEG results; equine samples should be stored for 30 minutes at RT.