Sun Y, Yiew XT, Beeler-Marfisi J
… +3 more, Kerr CL, Monteith G, Bateman S
Vet Clin Pathol
· 2025 Sep · PMID 40662223
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BACKGROUND: Regular blood sampling to monitor RBC mass in anemic cats can exacerbate anemia. Laboratory-based reporting can delay clinical decisions. A hand-held hemoglobinometer, HemoCue Hb 201 (HC-201), requires only o...BACKGROUND: Regular blood sampling to monitor RBC mass in anemic cats can exacerbate anemia. Laboratory-based reporting can delay clinical decisions. A hand-held hemoglobinometer, HemoCue Hb 201 (HC-201), requires only one drop of blood (10 μL) and provides results within 1 min. OBJECTIVES: This preliminary study aimed to evaluate the utility of HC-201 in cats and investigate the impact of potential interferents on its performance. METHODS: One hundred and fifty-four venous blood samples in EDTA from 93 cats were analyzed. Hemoglobin concentration was measured once using an ADVIA 2120 analyzer and compared to the mean of 2-3 replicate measurements from an HC-201. Agreement and systematic bias between HC-201 and ADVIA results, along with precision between HC-201 replicates, were assessed using Lin's concordance correlation coefficient, non-parametric Bland-Altman, Passing-Bablok regression, and intraclass correlation coefficient. The performance of HC-201 in the presence of anemia, leukocytosis, azotemia, lipemia, icterus, hemolysis, and peripheral versus jugular venipunctures was assessed using Wilcoxon rank-sum tests. RESULTS: Passing-Bablok analysis revealed a significant constant bias (intercept = -2.242, 95% CI: -4.042 to -0.667) but no significant proportional bias (slope = 1.015, 95% CI: 1.000-1.032). HC-201 demonstrated excellent agreement (ρ = 0.989) and precision (ICC = 0.997) with a median bias of -0.67 g/L (p = 0.001). The total observed error was 3.02%, within the allowable limits defined by international standards. Neither anemia, leukocytosis, azotemia, lipemia, nor venipuncture site influenced HC-201 measurements. Samples with icterus and hemolysis were insufficient in number for statistical comparison. CONCLUSIONS: This preliminary study indicates that HC-201 offers reliable point-of-care monitoring for hemoglobin concentration in cats.
Rout ED, Seymour K, Burnett R
… +3 more, Contreras C, Avery AC, Moore AR
Vet Clin Pathol
· 2025 Sep · PMID 40661054
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BACKGROUND: The PCR for Antigen Receptor Rearrangement (PARR) assay assesses clonality of a lymphoid population. Studies suggest PARR has decreased sensitivity for detecting clonality in canine plasma cell tumors (PCT) c...BACKGROUND: The PCR for Antigen Receptor Rearrangement (PARR) assay assesses clonality of a lymphoid population. Studies suggest PARR has decreased sensitivity for detecting clonality in canine plasma cell tumors (PCT) compared to lymphoma. OBJECTIVE: Assess sensitivity of PARR assays targeting the immunoglobulin heavy chain (IGH), immunoglobulin lambda light chain (IGL), and kappa deleting element (Kde) loci in canine PCT. METHODS: Canine cases included 35 PCTs (multiple myeloma and extramedullary, cutaneous or oral PCTs), 5 non-PCT foil cases, and 40 B-cell lymphoma cases. PCT diagnoses were confirmed via CD3, PAX5, and MUM1 immunolabeling, and some additionally had histopathology, serum or urine protein electrophoresis, or immunofixation. Nodal B-cell lymphomas were diagnosed by flow cytometry. Routine PARR (targeting IGH V-D-J and D-J rearrangements) and extended PARR (targeting additional IGH genes and IGL and Kde rearrangements) were performed on cytologic specimens (PCT and foil cases) and flow cytometry aspirates (lymphoma cases). Diagnostic sensitivity was calculated. RESULTS: Across both PARR assays, every PCT case with sufficient sample was interpreted as clonal and no foil cases were clonal. The routine PARR assay detected a clonal immunoglobulin rearrangement in 26/35 PCT cases (74.3%; 95% CI 56.7%-87.5%). The extended assay detected immunoglobulin gene clonality in 33/35 PCT cases (94.3%; 95% CI 80.8%-99.3%). 40/40 lymphoma cases were clonal in both the routine and extended PARR assays. CONCLUSION: The combined PARR assays used in this study, evaluating multiple IG loci, detected clonality in all PCTs tested, but routine PARR (targeting IGH) was less sensitive for detecting PCT clonality compared to B-cell lymphomas.
Vet Clin Pathol
· 2025 Dec · PMID 40657658
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BACKGROUND: Reference interval (RI) estimate inaccuracy is problematic at small sample sizes. Visual assessment of distribution histograms (VADH) may improve statistical technique selection, but its performance depends o...BACKGROUND: Reference interval (RI) estimate inaccuracy is problematic at small sample sizes. Visual assessment of distribution histograms (VADH) may improve statistical technique selection, but its performance depends on the human operator. OBJECTIVES: This study aimed to develop a machine learning model for small-sample VADH. METHODS: The training set consisted of 45 000 distribution histograms from samples ranging from 20 to 40 individuals extracted from simulated Gaussian, lognormal, and left-skewed populations. A convolutional neural network (CNN) was trained to predict the original population distribution by VADH. Its accuracy was tested on 900 human-classified histograms from a previous study (ranging from 20 to 60 individuals) and compared to the Shapiro-Wilk test performance in determining the original population distribution. A web application was developed to allow usage of the CNN, 95% RI estimation with 90% confidence intervals (CI) via bootstrapping, and outlier detection. RESULTS: The CNN model properly predicted the original population distribution by VADH in 84.0% (95% CI: 83.7-84.4) and 94.4% (95% CI: 92.7-95.8) of the samples of the training and the test sets, respectively. Comparatively, on the test set, the Shapiro-Wilk test accuracy was 65.0% (95% CI: 61.8-68.1) and 72.3% (95% CI: 69.3-75.2) using a p-value threshold of 0.05 and 0.2, respectively. The web application (named HARISS) was successfully deployed and is accessible at: https://hariss.streamlit.app/. CONCLUSIONS: The CNN model demonstrated effective VADH and might enhance RI estimate accuracy through the use of the HARISS web app, though proper reference individual selection and preanalytical/analytical conditions remain paramount.
Vet Clin Pathol
· 2025 Sep · PMID 40654288
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BACKGROUND: Thromboelastography (TEG) provides a global assessment of hemostasis and identifies patients with both hemorrhagic and thrombotic tendencies. Traditional TEG instruments, such as the TEG 5000 (Haemonetics), c...BACKGROUND: Thromboelastography (TEG) provides a global assessment of hemostasis and identifies patients with both hemorrhagic and thrombotic tendencies. Traditional TEG instruments, such as the TEG 5000 (Haemonetics), can be labor-intensive and require specialized training to operate. The TEG 6s (Haemonetics) is a commercially available point-of-care TEG that employs a cartridge-based system. Agreement between the TEG 6s and TEG 5000 is poor in dogs, and their viscoelastic measurements are not interchangeable. Reference intervals specific to the TEG 6s are needed for healthy adult dogs. OBJECTIVES: This study aimed to establish reference intervals for the TEG 6s in healthy adult dogs. METHODS: Citrated whole blood samples collected from 40 healthy adult dogs were analyzed using the TEG 6s and associated global hemostasis and lysis cartridges. Reference intervals were established using a nonparametric statistical method for all variables except for the Functional Fibrinogen level in the global hemostasis cartridge. RESULTS: Reference intervals were established for 12 TEG 6s measurands in healthy adult dogs. CONCLUSIONS: The results of this study provide reference intervals for TEG 6s variables associated with the global hemostasis and lysis cartridges in healthy adult dogs. Further studies are required to assess TEG 6s' utility in dogs with various disease states.
Font MB, Woolford L, Jaensch S
… +7 more, Hayward D, Hebart M, Dean E, Boardman WSJ, Stephenson T, Kovac J, Speight N
Vet Clin Pathol
· 2025 Sep · PMID 40600275
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BACKGROUND: Reference intervals (RIs) are an essential tool for assessment of clinical pathology data of animals, and are particularly important for monitoring the health status of free-ranging and captive wildlife, such...BACKGROUND: Reference intervals (RIs) are an essential tool for assessment of clinical pathology data of animals, and are particularly important for monitoring the health status of free-ranging and captive wildlife, such as koalas (Phascolarctos cinereus). OBJECTIVES: The purpose of this study was to: (1) provide comprehensive serum biochemistry reference intervals based on clinically healthy South Australian koalas from two populations, Mount Lofty Ranges (MLR) and Kangaroo Island (KI); and (2) identify any factors that can affect biochemical analytes, including koala retrovirus (KoRV) and Chlamydia pecorum subclinical infection status, age, sex, and population. METHODS: Serum biochemistry analytes were determined in 206 clinically healthy South Australian koalas caught from the wild in 2016 and 2018 using a Cobas 8000 Chemistry Analyzer and analyzed using Reference Value Advisor and SPSS v28 Statistical software. RESULTS: Biochemical reference intervals were established. Also, clinically and statistically significant differences in analytes were found based on age for alkaline phosphatase and phosphate, and albumin: globulin ratio, globulins, and total protein, most likely associated with physiological bone growth and immunological development, respectively, as observed in other species. Statistically significant differences between animals subclinically positive for KoRV and Chlamydia pecorum, were found for glucose and gamma glutamyl transferase respectively; however, these were marginal, and their reference intervals were similar. CONCLUSIONS: This study is the first to describe serum biochemical reference intervals for clinically healthy South Australian koalas of known Chlamydia and KoRV infection status. It represents an important tool to assist health assessments of koalas by veterinarians, as well as research and population monitoring.
Perego R, Spada E, Baggiani L
… +5 more, Ravasio G, Zucca E, Vanosi G, Ruffo G, Proverbio D
Vet Clin Pathol
· 2025 Jun · PMID 40521764
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BACKGROUND: To date, no studies have reported the evaluation of hemostatic activity in canine leukoreduced cryoprecipitate (LR-CRYO) and leukoreduced cryopoor plasma (LR-CPP). OBJECTIVES: We aimed to compare the hemostat...BACKGROUND: To date, no studies have reported the evaluation of hemostatic activity in canine leukoreduced cryoprecipitate (LR-CRYO) and leukoreduced cryopoor plasma (LR-CPP). OBJECTIVES: We aimed to compare the hemostatic activity of LR-CRYO and LR-CPP to leukoreduced fresh plasma (LR-FP) and to evaluate the preservation of LR-CRYO by refrigeration and refreezing after thawing. METHODS: Four hundred fifty milliliters of fresh blood was collected from ten donor dogs, leukoreduced, and separated into LR-FP, then frozen (-20°C) to obtain leukoreduced fresh frozen plasma (LR-FFP). LR-FFP was further separated into LR-CRYO and LR-CPP. LR-CRYO was frozen, thawed, and divided into two bags, one refrigerated for 24 h and one refrozen for 7 days. Factor VIII (FVIII) and X (FX) activity, prothrombin time, activated partial thromboplastin time, antithrombin III (ATIII) activity (ATA), total protein, albumin, fibrinogen, and D-dimer concentration, and von Willebrand Factor (vWF) activity were measured in LR-FP, LR-CRYO, LR-CPP, refrigerated, and refrozen LR-CRYO. RESULTS: FVIII activity was higher in LR-CRYO (p = 0.0001) versus LR-FP. vWF activity (p < 0.0001) and fibrinogen concentration (p = 0.0012) were lower in LR-CPP versus LR-FP. FX activity was higher in LR-CPP (p < 0.0001) and LR-FP (p = 0.0002) versus LR-CRYO, and albumin concentration was higher in LR-CPP versus LR-FP (p < 0.0001) and LR-CRYO (p < 0.0001). No statistically significant difference was found in refrigerated or refrozen LR-CRYO as compared with LR-CRYO, excluding ATA, which was lower (p = 0.0062) in refrigerated LR-CRYO. CONCLUSIONS: Because the concentration of FVIII is higher in LR-CRYO than in LR-FP, LR-CRYO is a possible component therapy when this factor is deficient. Since no statistically significant difference was found in refrozen LR-CRYO as compared with LR-CRYO, LR-CRYO can be frozen after thawing for reuse.
Moreira R, Ossowska M, Jornet-Rius O
… +2 more, Santos M, Cian F
Vet Clin Pathol
· 2025 Sep · PMID 40521731
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A 5-year-old neutered female Beagle was presented with lethargy, vomiting, and reduced appetite. Physical examination revealed mild hyperthermia and a cutaneous thickness on the right thigh. Hematology showed marked leuk...A 5-year-old neutered female Beagle was presented with lethargy, vomiting, and reduced appetite. Physical examination revealed mild hyperthermia and a cutaneous thickness on the right thigh. Hematology showed marked leukocytosis and moderate thrombocytopenia, consisting of 80% of atypical circulating cells, initially suggesting acute undifferentiated leukemia. Two weeks later, repeated hematology revealed unremarkable results. Cytology of the skin lesion showed a round cell neoplasia with features similar to the atypical circulating cells. Following the development of multiple cutaneous nodules and recurrence of clinical signs, further diagnostics, including flow cytometry, histopathology, and immunohistochemistry (IHC) of the skin nodules, as well as PCR for antigen receptor rearrangement (PARR) and immunocytochemistry (ICC) from the initial blood smear, confirmed a neoplastic T-cell proliferation consistent with cutaneous T-cell lymphoma with a probable transient leukemic phase. Despite chemotherapy, remission was short-lived, and the patient relapsed, ultimately leading to euthanasia. This case highlights a rare instance of transient leukemia likely originating from a primary cutaneous T-cell lymphoma, emphasizing the need for comprehensive diagnostic workups, combining hematology, biochemistry, cytology, flow cytometry, and immunophenotyping to avoid misclassification in hematologic malignancies.
Vet Clin Pathol
· 2025 Jun · PMID 40505004
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A 5-year-old spayed female golden retriever with a history of cutaneous asthenia (Ehlers-Danlos-like syndrome) was presented with a newly noted large hindlimb mass. Cytologic evaluation performed at the referring veterin...A 5-year-old spayed female golden retriever with a history of cutaneous asthenia (Ehlers-Danlos-like syndrome) was presented with a newly noted large hindlimb mass. Cytologic evaluation performed at the referring veterinarian was consistent with a keratinizing lesion. However, a second cytologic sample was obtained from the mass, aspirating from multiple aspects, and was concerning for a sarcoma. The mass was excised and submitted for histologic evaluation. The mass effect was diagnosed as a keratinizing cyst. Additionally, there were severe dermal collagen and elastin abnormalities with multifocal fibrosis and hemorrhage (consistent with cutaneous asthenia). To the best of the authors' knowledge, this is the first report of the cytologic findings in the skin of a canine with cutaneous asthenia.
Vet Clin Pathol
· 2025 Jun · PMID 40476643
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BACKGROUND: Neutrophil cell population data (CPD), including fluorescent light intensity (NE-SFL) and side scatter (NE-SSC), are promising inflammatory markers in human sepsis but remain unexplored in dogs and cats. OBJE...BACKGROUND: Neutrophil cell population data (CPD), including fluorescent light intensity (NE-SFL) and side scatter (NE-SSC), are promising inflammatory markers in human sepsis but remain unexplored in dogs and cats. OBJECTIVES: Determine the diagnostic utility of NE-SSC and NE-SFL for detecting systemic inflammation in dogs and cats. METHODS: Dogs and cats with archived CPD, blood films, and acute phase protein (APP) measurements were included. Increased C-reactive protein (CRP) in dogs and Serum Amyloid A (SAA) in cats were considered indicative of systemic inflammation. CPD was compared with APPs, white cell count (WCC), neutrophil count, band neutrophil count, and toxic change grade. Optimal cut-offs and associated sensitivities and specificities were calculated using ROC curve analysis. Correlations were assessed using Spearman's coefficient. RESULTS: NE-SFL and NE-SSC were significantly increased in dogs and cats with systemic inflammation. The area under the curve (AUC) of NE-SFL was higher than that of NE-SSC, WCC, and band neutrophil count in both dogs (0.82) and cats (0.77). The optimal NE-SFL cut-off for detecting systemic inflammation was > 41.7 ch in dogs (sensitivity 80%; specificity 66%) and > 37.4 ch in cats (sensitivity 75%; specificity 67%). NE-SFL was positively correlated with APPs, WCC, neutrophil count, and band neutrophil count in both species. NE-SSC was positively correlated with APPs in both species and, in dogs, also with WCC, neutrophil count, and band neutrophil count. CONCLUSION: CPD, particularly NE-SFL, is a promising marker of inflammation in dogs and cats and could be especially useful when APP quantification or blood smear examination are unavailable.
Rautenbach Y, Meyer LCR, Goddard A
… +2 more, Buss PE, Hooijberg EH
Vet Clin Pathol
· 2025 Jun · PMID 40464770
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BACKGROUND: Skeletal and possible cardiac muscle damage has been reported in chemically immobilized and transported African rhinoceros during conservation-related activities. The extent of cardiac muscle injury in these...BACKGROUND: Skeletal and possible cardiac muscle damage has been reported in chemically immobilized and transported African rhinoceros during conservation-related activities. The extent of cardiac muscle injury in these rhinoceros is unknown due to a lack of validated cardiac troponin I (cTnI) assays. However, recently, five human cTnI assays were deemed suitable for analytical validation in African rhinoceros based on cTnI sequencing results. OBJECTIVES: The first objective was to validate two cTnI immunoassay point-of-care analyzers (POCAs) in African rhinoceros and, secondly, to perform quality control (QC) validation for the POCAs. METHODS: Analytical validation of the Stratus CS Acute Care Troponin I cTnI immunoassay and Atellica VTLi high sensitivity cTnI (hs-cTnI) assay was performed using rhinoceros serum samples and species-specific cardiac muscle homogenate. Experiments included precision studies, reportable range, hemoglobin interference studies, recovery studies, and detection limit studies, with results assessed against prescribed total allowable error (TE) performance goals. Commercial quality control material (QCM) data were used to calculate bias and imprecision for QC validation. RESULTS: Imprecision was acceptable (1.9%-10.3%) and met low cTnI concentration performance goals. Reportable ranges were similar to the manufacturer's specifications. High hemoglobin concentrations in white rhinoceros resulted in a positive bias in the Stratus CS. A simple 1 QC rule using two levels of QCM and a TEa of 70% could be used in both analyzers, except at very low cTnI concentrations in the Atellica VTLi. CONCLUSIONS: Both cTnI POCAs are suitable for use in African rhinoceros, and analytical performance goals for low cTnI concentrations in hs-cTnI assays were met.
Legroux D, Kersten L, Barral G
… +3 more, Mauras A, Buronfosse T, Ramery E
Vet Clin Pathol
· 2025 Jun · PMID 40462459
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BACKGROUND: The presence of both nucleated RBC and thrombocytes in birds makes the determination of hematological variables difficult. This leads to time-consuming manual evaluation. New-generation analyzers are hypothes...BACKGROUND: The presence of both nucleated RBC and thrombocytes in birds makes the determination of hematological variables difficult. This leads to time-consuming manual evaluation. New-generation analyzers are hypothesized to be both time-saving and accurate. OBJECTIVES: The objective was to evaluate the performance of two hematology analyzers in 27-day-old broilers and to establish reference intervals (RIs) for RBC and WBC variables obtained by automated and manual methods. METHODS: Automated HCT, WBC count, and differentials obtained using Sysmex XT-2000iV and XN-1000V on 309 and 56 EDTA blood samples from Ross 308 male broilers, respectively, were compared to manual PCV, manual WBC count, and differentials from November 2021 to November 2022. RIs for automated HCT and RBC variables (from impedance channel) on one hand, and manual PCV and WBC's variables on the other hand, were established. RESULTS: The Sysmex XT-2000iV was accurate for HCT and outperformed the XN-1000V but, both automated methods and manual methods cannot be considered interchangeable. Constant error, proportional error, and bias were identified and calculated for each analyzer for HCT. Both analyzers were unable to enumerate WBC. RIs were established as follows: HCT-XT (L/L) [0.27-0.36]; HCT-XN (L/L) [0.24-0.38]; manual PCV (L/L) [0.27-0.37]; manual WBC (×10/L) [10.0-38.4]; manual differential (×10/L) with heterophils [1.5-16.2]; lymphocytes [5.1-18.9]; monocytes [0.0-2.1]; eosinophils [0.0-2.2]; basophils [0.0-3.1]. CONCLUSIONS: Both analyzers are usable for the evaluation of HCT in Ross 308 chickens and in specific conditions for the other RBC variables. Manual methods are still preferred for the evaluation of WBC variables.
Nikvand AA, Jalali SM, Bahrami S
… +1 more, Rahij Torfi H
Vet Clin Pathol
· 2025 Jun · PMID 40462454
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Trypanosoma evansi (T. evansi) is a hemoprotozoan parasite affecting camels and equids, such as horses, mules, and donkeys, and is known to cause surra disease in these animals. Despite the worldwide distribution of T. e...Trypanosoma evansi (T. evansi) is a hemoprotozoan parasite affecting camels and equids, such as horses, mules, and donkeys, and is known to cause surra disease in these animals. Despite the worldwide distribution of T. evansi infections in equids, surra has not been reported in Arabian horses in Khuzestan Province for over 60 years. In September 2018, a 7-year-old Arabian mare was referred from a 10-horse farm in the suburbs of Ahvaz City. The mare presented with a history of weight loss, poor appetite, and proximity to a camel herd. Physical examination revealed a poor body condition score (BCS ≤ 3/9), pale conjunctival and oral mucosal membranes, dehydration, tachycardia, and fever. Other horses in the herd also exhibited gradual weight loss and hyporexia, with two fatalities reported. A hematologic examination of the referred mare showed leukopenia, severe anemia, and thrombocytopenia. Microscopic examination of a whole blood smear confirmed severe infection with Trypanosoma. PCR assessment of a blood sample further confirmed the infection with T. evansi. The referred mare and seven nonreferred patients from the herd received treatment with diminazene diaceturate (5 mg/kg, a single dose) and penicillin for 4 days. Unfortunately, the referred mare died despite the treatment, while the remaining nonreferred horses recovered. This report marks the first molecular diagnosis of surra in an Arabian horse in Khuzestan Province, highlighting the need for further investigation into surra as a re-emerging disease in this significant population of Arabian horses in the region.
Harada H, Tamura M, Ishii K
… +2 more, Osuga T, Ohta H
Vet Clin Pathol
· 2025 Jun · PMID 40462420
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BACKGROUND: Pancreas-specific lipase and lipase activity measured by immunological assays (Spec cPL and Spec fPL) and colorimetric assays (1,2-o-dilauryl-rac-glycelo-3-glutaric acid-(6'-methylresorufin) ester [DGGR] and...BACKGROUND: Pancreas-specific lipase and lipase activity measured by immunological assays (Spec cPL and Spec fPL) and colorimetric assays (1,2-o-dilauryl-rac-glycelo-3-glutaric acid-(6'-methylresorufin) ester [DGGR] and triolein), respectively, are used to diagnose pancreatitis in both dogs and cats. However, DGGR and triolein assays may be influenced by extrapancreatic lipases, including hepatic triglyceride lipase (HTGL) and lipoprotein lipase (LPL). OBJECTIVES: To investigate the effect of extrapancreatic lipases on immunological and colorimetric assays by measuring changes in HTGL and LPL activity following heparin administration. METHODS: Six healthy Beagles and six adult purpose-bred cats were enrolled. HTGL and LPL activities were induced by intravenous heparin administration. Serum samples were collected at baseline and at 5-, 10-, 15-, and 60-min following heparin injection. Spec cPL, Spec fPL, and lipase activities were measured using DGGR and triolein assays, whereas HTGL and LPL activities were measured using their respective assays. RESULTS: Spec cPL and Spec fPL levels showed no significant changes following heparin administration. Conversely, DGGR-based and triolein-based lipase activities, as well as HTGL and LPL activities, were significantly increased after heparin administration in both dogs and cats. HTGL and LPL activities showed significant positive correlations with DGGR-based (P < .001, r = .90 for both) and triolein-based (P < .001, r = .63 and P < .001, r = .68, respectively) lipase activities, but not with Spec cPL and Spec fPL. CONCLUSIONS: DGGR- and triolein-based lipase activities are influenced by HTGL and LPL activities, as their substrates are hydrolyzed by pancreatic lipase, HTGL, and LPL.
Vet Clin Pathol
· 2025 Dec · PMID 40462415
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Artificial intelligence (AI), particularly through machine learning and deep learning, presents opportunities for the enhancement of the workflow of the veterinary clinical pathologist. This review introduces basic conce...Artificial intelligence (AI), particularly through machine learning and deep learning, presents opportunities for the enhancement of the workflow of the veterinary clinical pathologist. This review introduces basic concepts in AI in a nontechnical manner and explores the qualification and integration of AI in veterinary clinical pathology. The veterinary clinical pathologist must play an active role in defining the intended use, design, and qualification of these methods as well as the plan for monitoring their responsible application in practice.