Bou P, Mesa I, Ferreira RF
… +3 more, Torrente C, Manzanilla EG, Bosch L
Vet Clin Pathol
· 2024 Dec · PMID 39307821
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BACKGROUND: Packed RBC (pRBC) transfusions are often necessary to enhance organ perfusion and tissue oxygenation in cases of severe anemia. OBJECTIVES: We aimed to describe changes in acid-base and biochemical parameters...BACKGROUND: Packed RBC (pRBC) transfusions are often necessary to enhance organ perfusion and tissue oxygenation in cases of severe anemia. OBJECTIVES: We aimed to describe changes in acid-base and biochemical parameters in dogs after transfusion of pRBC and potential effects on the outcome. METHODS: The prospective observational study included anemic dogs requiring pRBC transfusions. Venous blood gas and electrolytes were measured pre-transfusion and repeated within 1 h post-transfusion. Signalment, pre-existing conditions and underlying disease(s), number of days spent of hospitalization, the quantity of transfused pRBC units, and the outcome were also recorded. Associations between analytical values and all other parameters before and after transfusion were assessed using the Wilcoxon test. A generalized linear model was generated for each of the blood gas parameters post-transfusion, including the pre-transfusion values and the characteristics of the animal as fixed effects. The effect of the blood gas parameters on the mortality and hospitalization length was assessed using logistic regression and generalized linear models, respectively. RESULTS: Twenty-six dogs with different causes of anemia were included in the study, with dogs remaining hospitalized for a median of 3 days (range: 0-11). Pre-transfusion heart rate, respiratory rate, Angap, base excess, and lactate were higher than post-transfusion values. Post-transfusion results showed an increase in hematocrit, hemoglobin, HCO-, PCO, tCO, and chloride. Pre-transfusion hypophosphatemia and receiving an increased volume of transfused pRBC were associated with a longer hospital stay, while higher pre-transfusion lactate values were associated with higher mortality. CONCLUSIONS: pRBC transfusion in anemic dogs was associated with an improvement in perfusion and acid-base parameters. Pre-transfusion lactate may be an indicator of mortality in dogs.
García R, Pastor J, de la Fuente C
… +1 more, Añor S
Vet Clin Pathol
· 2024 Dec · PMID 39307760
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BACKGROUND: Thromboelastography (TEG) is an effective technique to assess the efficiency of coagulation. Phenobarbital (PB) can induce hematological and coagulation disorders in both animals and humans, but its effects o...BACKGROUND: Thromboelastography (TEG) is an effective technique to assess the efficiency of coagulation. Phenobarbital (PB) can induce hematological and coagulation disorders in both animals and humans, but its effects on hemostasis have been little investigated and are poorly understood in dogs. OBJECTIVES: The aim of this article was to assess coagulation using TEG in a population of dogs with idiopathic epilepsy treated with PB. MATERIAL AND METHODS: Prospective observational study. TEG was performed in blood samples from dogs with idiopathic epilepsy that were divided in three groups: Two groups of treated dogs that were on phenobarbital treatment for less or more than 6 months, and a control group of healthy dogs. Duration of treatment, dose, phenobarbital serum concentration and selected hematological and biochemical parameters were evaluated and correlated with the TEG results. RESULTS: No statistically significant differences were found between groups. None of the animals appeared to be in a hypo- or hypercoagulable state, however 9/19 (47,4%) dogs were classified as hyper-fibrinolytic. A statistically significant negative relationship between MA and G values and increased fibrinolytic activity (LY30) were found. No statistically significant relationship was found between PB dose or PB blood levels and TEG parameters in either group. No bleeding complications were observed. DISCUSSION: The rise in fibrinolysis might be due to hepatic damage from PB, as indicated by elevated liver enzymes in many dogs with abnormal fibrinolytic patterns. Although TEG showed hyperfibrinolysis in some dogs, the presence of primary or secondary hyperfibrinolysis could not be confirmed due to the lack of D-dimer measurements and liver biopsy. TEG's sensitivity compared to other fibrinolysis biomarkers like PAP might also affect results. CONCLUSIONS: TThe cause of hyperfibrinolysis in epileptic dogs treated with phenobarbital remains unclear, with potential links to hepatic effects or handling, and further research is needed to assess its clinical significance.
BACKGROUND: The Sysmex XN-V is derived from the new Sysmex XN series of human hematology analyzers. The main changes from the previously validated XT-2000iV analyzer include an optic-fluorescent analysis for platelets an...BACKGROUND: The Sysmex XN-V is derived from the new Sysmex XN series of human hematology analyzers. The main changes from the previously validated XT-2000iV analyzer include an optic-fluorescent analysis for platelets and a nucleated red blood cell (NRBC) count. OBJECTIVE: We aimed to validate the Sysmex XN-V for feline blood following the American College for Veterinary Clinical Pathology and International Council for Standardization in Hematology recommendations. METHODS: Feline EDTA blood specimens were analyzed on the Sysmex XN-V to evaluate repeatability, linearity, comparison with the XT-2000iV analyzer and manual methods, stability, and to verify the previously established Sysmex XT-2000iV RIs. RESULTS: Repeatability was excellent for most variables. Visually determined linearity was excellent or good for most variables except eosinophils and platelet variables. The correlation between the XN-V and XT-2000iV analyzers was good (≥0.82) for all variables except reticulocyte indices. Correlations between the Sysmex XN-V and manual differential counts were good to excellent for most variables, acceptable for neutrophils, and fair for monocytes and NRBC. The previously established Sysmex XT-2000iV RIs can be used to interpret results from the Sysmex XN-V analyzer for most variables except red cell distribution width and reticulocyte variables. The RI for platelet variables could not be evaluated because of platelet clumps. Changes in the Sysmex XN-V measurements after storage at 4 and 24°C were similar to those described for the Sysmex XT-2000iV analyzer. CONCLUSIONS: The performance of the Sysmex XN-V analyzer was good and compared favorably with the Sysmex XT-2000iV analyzer.
Rogato F, Szladovits B, Martineau H
… +5 more, Yale AD, Mitchell JL, Holmes EJ, Gardner RH, Guillén A
Vet Clin Pathol
· 2024 Dec · PMID 39289176
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An eight-year-old male neutered crossbreed dog presented with erosive and ulcerative cutaneous lesions in the inguinal regions, the medial aspect of both thighs, and the stifles. Hematologic assessment revealed nonregene...An eight-year-old male neutered crossbreed dog presented with erosive and ulcerative cutaneous lesions in the inguinal regions, the medial aspect of both thighs, and the stifles. Hematologic assessment revealed nonregenerative anemia, thrombocytopenia, and high numbers of neoplastic mononuclear cells with a variable degree of maturation. The mononuclear neoplastic cells, with nuclei measuring 10-20 microns in diameter, accounted for 57% of the nucleated blood cells. In addition, the blood contained increased numbers of mature neutrophils and monocytes with atypical morphology. Cytologic examination of the right popliteal lymph node found high numbers of large mononuclear cells with similar morphology to those in the peripheral blood. Flow cytometry of peripheral blood revealed expression by the mononuclear neoplastic cells of the pan-leukocyte marker CD45 and myeloid markers CD14, MAC387, and myeloperoxidase (MPO). These results confirmed a diagnosis of acute myeloid leukemia (AML). Computed tomography found moderate nodular hepatosplenomegaly and multifocal bi-cavitary lymphadenopathy. Histopathologic examination of biopsies from the cutaneous lesions identified infiltration of the dermis by intermediate to large neoplastic round cells. Further treatment was declined, and the owners elected euthanasia. Postmortem examination confirmed AML involvement in the bone marrow, peripheral and intracavitary lymph nodes, heart, liver, kidney, and skin. Neoplastic cells in the bone marrow and skin showed positive immunolabeling for ionized calcium-binding adaptor protein 1 and MPO. To the best of our knowledge, this is the first report of ulcerative cutaneous lesions observed among the presenting clinical signs in a dog with AML and secondary leukemia cutis.
BACKGROUND: Regression describes the relationship of results from two analyzers, and the generated equation can be used to harmonize results. Point-of-care (POC) analyzers cannot be calibrated by the end user, so regress...BACKGROUND: Regression describes the relationship of results from two analyzers, and the generated equation can be used to harmonize results. Point-of-care (POC) analyzers cannot be calibrated by the end user, so regression offers an opportunity for calculated harmonization. Harmonization (uniformity) of laboratory results facilitates the use of common reference intervals and medical decision thresholds. OBJECTIVE: Our aims were to characterize the relationship of results for multiple biochemistry analytes on a POC and a commercial laboratory analyzer (CL) with three regression techniques and to use regression equations to harmonize the POC results with those of the CL. Harmonized results were assessed by recognized quality goals. We used harmonized results to assess the regression techniques. METHODS: After analyzer imprecision assessments, paired clinical samples were assessed with one dataset to calculate regression parameters that were applied to a second dataset. Three regression techniques were performed, and each was used to harmonize the POC results with those from the CL. POC results were assessed for bias and the number of results reaching quality goals before and after harmonization. RESULTS: All regression techniques could be used to harmonize most analytes so that 95% of results were within ASVCP TEa guidelines. Harmonization could be further improved with alternate regression techniques or exclusions. CONCLUSIONS: Regression offers a means to harmonize POC and CL analyzers. Further work is needed to assess how few samples can reliably be used and to assess likely species differences. No regression technique reliably describes the relationship between methods when correlation is poor.
During a 3-year time period, a 15-year-old male castrated Terrier mix (dog 1) and a 6-year-old female spayed Labrador Retriever (dog 2) presented to the North Carolina State Veterinary Hospital with similar blood work ab...During a 3-year time period, a 15-year-old male castrated Terrier mix (dog 1) and a 6-year-old female spayed Labrador Retriever (dog 2) presented to the North Carolina State Veterinary Hospital with similar blood work abnormalities and no significant physical examination findings. A CBC, chemistry panel, and urinalysis performed on both dogs were relatively unremarkable, other than a marked increase in serum gamma-glutamyltransferase (GGT) activity. Through imaging, both patients were diagnosed with a renal mass, and histopathology of both masses revealed a carcinoma. Immunohistochemical staining of the renal mass in both dog 1 and dog 2 were intensely positive for GGT. Dog 1 had the affected kidney removed, which normalized the GGT value. Dog 2 was euthanized, and metastasis to the lung was noted upon postmortem examination. There have been limited case studies documenting an elevation in serum GGT in dogs diagnosed with renal carcinoma. While renal carcinoma is uncommon in dogs, it is an important differential to keep in mind when there is a marked increase in serum GGT without accompanying increases in other measured liver enzymes. In addition, serum GGT can serve as a helpful biomarker for disease resolution and recurrence, as surgical removal of the renal mass (dog 1) led to the resolution of the elevated serum GGT. To our knowledge, this is the first report demonstrating IHC staining for GGT in a canine renal carcinoma.
BACKGROUND: Paraoxonase-1 (PON-1) has been suggested as a marker of inflammation and oxidative stress in horses and could potentially be used for prognostication in horses with colitis. OBJECTIVES: Assessment of PON-1 in...BACKGROUND: Paraoxonase-1 (PON-1) has been suggested as a marker of inflammation and oxidative stress in horses and could potentially be used for prognostication in horses with colitis. OBJECTIVES: Assessment of PON-1 in horses with colitis and comparison of two methods. METHODS: Serum PON-1 was measured by two methods (paraoxon and p-nitrophenyl acetate) in 161 horses with colitis and 57 controls. Follow-up samples obtained during hospitalization were available from 106 horses with colitis. The two methods were compared. RESULTS: Serum PON-1 was significantly lower in horses with colitis than in healthy horses (P < .0001 for both methods) as well as in nonsurvivors compared with survivors (P = .0141 [paraoxon-based method] and P < .0001 [p-nitrophenyl acetate-based method]), but with marked overlap between groups. PON-1 activity did not change parallel to a change in inflammatory status in response to treatment when assessed at admission and in up to seven follow-up samples. Admission PON-1 activity could not reliably classify horses as survivors or nonsurvivors, with sensitivity and specificity ranging between 53.1% and 72.9%. Results from the two methods were comparable. CONCLUSIONS: Both methods reliably measured serum PON-1 activity. Significant differences in PON-1 activity were found between healthy horses and horses with colitis and between survivors and nonsurvivors. However, PON-1 activity varied considerably within groups. Both the proposed reference intervals as well as alternative cutoff values resulted in suboptimal diagnostic and prognostic performance, and the use of serum PON-1 in horses with colitis thus seems to add little to existing diagnostic and prognostic markers.
A 7-year-old female spayed Bernese Mountain dog was presented for evaluation of hematuria. Incidentally, a right stifle sarcoma was diagnosed via cytology, which raised concern for histiocytic sarcoma (given the patient'...A 7-year-old female spayed Bernese Mountain dog was presented for evaluation of hematuria. Incidentally, a right stifle sarcoma was diagnosed via cytology, which raised concern for histiocytic sarcoma (given the patient's signalment) versus another joint-associated sarcoma. Histopathology and immunohistochemistry revealed a CD18-negative, non-histiocytic origin cell population. Findings were consistent with a joint-associated grade II soft tissue sarcoma (STS). The patient's hematuria was progressive over 5 months, and urinary bladder transitional cell carcinoma (TCC) was diagnosed via cystoscopy and histopathology. An enlarged right medial iliac lymph node was identified on routine restaging via abdominal ultrasound 3 months later. Cytology of the lymph node revealed a markedly pleomorphic cell population, again raising concern for histiocytic sarcoma (HS). Other differentials included an anaplastic metastatic population from the joint-associated STS or the TCC. Immunocytochemistry revealed a cytokeratin-positive, CD18-, CD204-, and vimentin-negative cell population, consistent with a carcinoma. DNA was extracted from cytology slides to sequence cells for BRAF mutation status. Sequencing revealed a homozygous V596E (transcript ENSCAFT00845055173.1) BRAF mutation, consistent with the known biology of TCC. In neither case was HS truly present in this patient, but immunocytochemistry provided information that helped to optimize the patient's chemotherapy recommendations.
A 3-year-old male neutered domestic shorthair cat presented with lethargy, hyporexia, and pyrexia of unknown origin. Biochemical analysis using a Beckman Coulter AU480 demonstrated marked increases in creatine kinase and...A 3-year-old male neutered domestic shorthair cat presented with lethargy, hyporexia, and pyrexia of unknown origin. Biochemical analysis using a Beckman Coulter AU480 demonstrated marked increases in creatine kinase and aspartate aminotransferase, indicative of severe muscle injury, with concurrent presumptive myoglobinuria on urinalysis. A marked, non-physiologic increase in measured bicarbonate and resultant negative anion gap was documented; however, calculated bicarbonate obtained via a point-of-care blood gas analyzer was within normal limits. Laboratory error due to interference by lactate dehydrogenase was suspected and supported by the results of subsequent biochemical testing. Artifactual increases in bicarbonate have been documented in cases of rhabdomyolysis in horses, cows, and a bird. However, to the best of the authors' knowledge, this is the first report to demonstrate this spurious change in a cat.
Peters LM, Reding Graf T, Giori L
… +3 more, Mevissen M, Graf R, Howard J
Vet Clin Pathol
· 2024 Jun · PMID 38825585
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BACKGROUND: Regenerating island-derived proteins (REG) are upregulated in people with sepsis, pancreatitis, and gastrointestinal diseases. One member of the REG family, namely REG3E, was recently identified in pancreatic...BACKGROUND: Regenerating island-derived proteins (REG) are upregulated in people with sepsis, pancreatitis, and gastrointestinal diseases. One member of the REG family, namely REG3E, was recently identified in pancreatic tissue and plasma of dogs, with high expression in pancreatitis and sepsis. OBJECTIVES: We aimed to develop and validate an ELISA to measure REG3E concentrations in canine blood. METHODS: An indirect sandwich ELISA was developed using recombinant canine REG3E protein and polyclonal anti-canine REG3E antibodies raised in guinea pigs and rabbits. Antibody specificity was assessed using western blot and mass spectrometric analysis of protein purified from canine plasma. Assay validation included evaluation of dilutional linearity, parallelism, spiking recovery, repeatability and reproducibility, stability, interferences, and comparison of serum and heparinized plasma. RESULTS: Antibodies bound specifically to REG3E with no evidence of cross-reactivity with other proteins. The limit of detection of the ELISA was 15 ng/mL, and the lower limit of quantification was 30 ng/mL. The assay demonstrated good to excellent linearity, dilutional and mixing parallelism, and recovery, with mean observed-to-expected ratios of 104%, 107%, 102%, and 92%, respectively, and no evidence of a hook effect. Coefficients of variation were ≤8.5% for repeatability and ≤14.3% for reproducibility at three different levels. Measurements of REG3E in plasma were not significantly influenced by different storage conditions, freeze-thawing cycles, hemolysis, lipemia, or icterus. There was no significant difference between REG3E concentrations in heparinized plasma and serum samples. CONCLUSIONS: The canine REG3E ELISA has acceptable precision, accuracy, linearity, and reproducibility for the measurement of REG3E in canine plasma and serum.
Nielsen LN, Petersen MB, Capion N
… +2 more, Lundsgaard JF, Jensen AL
Vet Clin Pathol
· 2024 Jun · PMID 38825583
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BACKGROUND: Acute phase proteins are a group of vital constituents of the innate immune system, which may also serve as circulatory biomarkers of inflammation. The major acute phase protein serum amyloid A (SAA) is a rel...BACKGROUND: Acute phase proteins are a group of vital constituents of the innate immune system, which may also serve as circulatory biomarkers of inflammation. The major acute phase protein serum amyloid A (SAA) is a reliable and sensitive biomarker in cows, allowing for rapid detection of inflammatory disease. A multispecies automated immunoturbidimetric assay (VET-SAA, Eiken) has been validated for horses, dogs, and cats, and it has been used to measure SAA concentrations in bovine samples. OBJECTIVES: The aim of the present study was to perform an analytical validation of the VET-SAA immunoturbidometric assay based on monoclonal antihuman SAA antibodies for the measurement of SAA in clinical samples from cows. METHODS AND RESULTS: The validation included an assessment of imprecision, inaccuracy, and detection limit, as well as an evaluation of the overlap performance, using banked serum from healthy and sick cows with or without inflammatory disease. Intra- and interassay variation ranged from 0.91% to 2.9% and 2.5% to 5.8%, respectively. The assay was performed with acceptable accuracy within a clinically relevant range of SAA, although minor signs of inaccuracy were detected. Overlap performance was acceptable, with the VET-SAA assay able to differentiate between healthy cows and cows with inflammatory and noninflammatory conditions. The automated VET-SAA assay is considered acceptable for the measurement of SAA in cows.
Akiyoshi M, Hisasue M, Goto-Koshino Y
… +4 more, Asakawa MG, Neo S, Akiyoshi M, Tomiyasu H
Vet Clin Pathol
· 2024 Jun · PMID 38806404
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A 12-year-old spayed female Dalmatian presented with acute vomiting and anorexia. The clinicopathological and imaging abnormalities included icterus, biliary obstruction, and multiple diffuse splenic hypoechogenic nodule...A 12-year-old spayed female Dalmatian presented with acute vomiting and anorexia. The clinicopathological and imaging abnormalities included icterus, biliary obstruction, and multiple diffuse splenic hypoechogenic nodules. Cholecystectomy was performed to remove the obstruction, followed by liver biopsy and splenectomy. Histopathological and immunohistology evaluation of the spleen, liver, and gallbladder revealed splenic marginal zone lymphoma (MZL) with gallbladder and hepatic infiltration of neoplastic CD20/CD79α-positive cells. Moreover, we observed clonal rearrangements of the immunoglobulin heavy-chain (IgH) gene in all three tissues. The dog was in good condition without chemotherapy. However, there was progressive elevation of liver enzymes, which could be attributed to neoplastic hepatic infiltration. Chlorambucil and prednisolone were administered until day 108, when the liver enzyme levels normalized. On day 156, the dog developed diffuse large B-cell lymphoma (DLBCL) of the peripheral lymph nodes. Sequence analysis of the clonally rearranged IgH gene revealed that all neoplastic cells in the spleen, gallbladder, and liver at initial presentation, as well as lymph nodes on day 156, possessed the same sequence identity of the amplified IgH fragments. This demonstrated that all neoplastic cells were derived from the same B-lymphocyte clone. The DLBCL was considered to have transformed from the splenic MZL, with gallbladder involvement. In cases of splenic MZL, it is important to consider gallbladder involvement and transformation to DLBCL. Moreover, gallbladder lymphoma should be included in the differential diagnosis of dogs with gallbladder abnormalities. Further studies are warranted to investigate the prognosis of splenic MZL.
Tamamoto T, Miki Y, Sakamoto M
… +6 more, Yoshii M, Yamada M, Sudo D, Fusato Y, Ozawa J, Satake C
Vet Clin Pathol
· 2024 Jun · PMID 38803017
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BACKGROUND: Biochemical measurements are commonly evaluated using population-based reference intervals; however, there is a growing trend toward reassessing results with within-subject variation (CV). OBJECTIVES: We aime...BACKGROUND: Biochemical measurements are commonly evaluated using population-based reference intervals; however, there is a growing trend toward reassessing results with within-subject variation (CV). OBJECTIVES: We aimed to estimate the CV of 16 biochemical analytes using a large database of dogs and cats, which refers to the results of routine health checkups. METHODS: Pairs of sequential results for 16 analytes were extracted from a database of adult patients. The second result was divided by the first result to produce the ratio of sequential results (rr), and the frequency distribution of rr was plotted. From the plots, the coefficient of variation (CV) was calculated. Analytical variation (CV) was calculated using quality control data, and CV was estimated as follows: . Estimated CV was compared with previously reported CV using the Bland-Altman plot analysis. RESULTS: From the database, 9078 data points from 3610 dogs and 3743 data points from 1473 cats were extracted, with 5468 data pairs for dogs and 2270 for cats. Sampling intervals ranged from 10 to 1970 days (median 366) for dogs and 23 to 1862 days (median 365) for cats. Bland-Altman analysis showed most CV plots fell within the limits of agreement; however, positive fixed biases were observed in both dogs and cats. CONCLUSIONS: Our study introduces a novel approach of estimating CV using routine health checkup data in dogs and cats. Despite biases, our method holds promise for clinical application in assessing the significance of measurement result differences.
Cagle LA, Casal M, Agnew D
… +3 more, Skinner S, Lanier CJ, Harvey JW
Vet Clin Pathol
· 2024 Jun · PMID 38797715
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A 13-year-old male domestic short-hair cat presented for evaluation of labored breathing, hyporexia, and lethargy. Pertinent initial diagnostics yielded leukocytosis, characterized by neutrophilia and monocytosis. Numero...A 13-year-old male domestic short-hair cat presented for evaluation of labored breathing, hyporexia, and lethargy. Pertinent initial diagnostics yielded leukocytosis, characterized by neutrophilia and monocytosis. Numerous small, round, magenta granules were observed within all neutrophils in Wright-Giemsa-stained blood films on the day of presentation and the day thereafter. No other neutrophil morphologic abnormalities were present, making cytoplasmic toxicity highly unlikely. Hyperadrenocorticism was diagnosed based on the lack of suppression in a low-dose dexamethasone suppression test, and without other diagnostics, the cat was discharged on trilostane therapy. Neutrophil granules did not stain with Alcian blue pH 1.0, periodic acid-Schiff (PAS), PAS and Alcian blue pH 2.5, and toluidine blue. Electron microscopy identified no differences in the morphology of the secretory granules or other neutrophil features. Metabolic screening tests of the cat's urine did not identify a genetic metabolic disorder. However, serum α- and β -hexosaminidase (HexA and HexB) activities were 4.3% and 0% of normal controls, respectively, which is supportive of GM2-gangliosidosis, that is, Sandhoff disorder. However, the historical, clinical, and electron microscopy findings did not provide evidence to confirm this genetic defect. To the author's knowledge, this is the first case of magenta-staining granules within neutrophils in a breed other than a Birman, Siamese, or Himalayan.