Evaluation of point-of-care analysers for blood gas and clinical chemistry in Hermann's tortoises (Testudo hermanni).

OBJECTIVE: To assess the agreement between point-of-care and laboratory analysers in measuring biochemical and blood gas analytes in venous samples from tortoises and to define preliminary reference intervals for venous blood gas analysis in Hermann's tortoises (Testudo hermanni). MATERIALS AND METHODS: Jugular venous blood samples from 47 Hermann's tortoises underwent paired analysis with a portable gas analyser (i-STAT 1, Abaxis), a portable chemical analyser (VetScan VS2, Abaxis), and with the respective reference analysers. Agarose gel electrophoresis was used to determine albumin concentrations on 12 specimens. Agreement was evaluated with Bland-Altman plots and regression analysis using the Passing-Bablok method. RESULTS: Point-of-care analysers had variable agreement with the reference analysers, presenting constant or proportional bias depending on the analyte. Relevant analytes in reptiles, such as ionised and total calcium, had acceptable agreement. The method for determining albumin concentration currently available in both point-of-care and laboratory analysers significantly overestimated albumin concentrations as compared to protein electrophoresis. CLINICAL SIGNIFICANCE: While the use of POC analysers is extremely advantageous in small animal primary care facilities, agreement between point-of-care and laboratory analysers varies depending on the analyte. For certain analytes, interchangeability of results is limited and specific reference intervals for point-of-care analysers are required. Veterinarians should be aware of the size and the direction of the bias of each analyte.

Evaluation of 5 methods for diagnosing failure of passive transfer in 160 Holstein calves.

BACKGROUND: Inadequate absorption of colostral IgG1 is termed failure of transfer of passive immunity (FTPI). Dairy calves with FTPI have increased mortality and morbidity in their first 6 months of life. OBJECTIVES: This study compared the clinical performance of 5 methods for diagnosing FTPI in Holstein calves. METHODS: An observational study was performed using 160 Holstein heifer calves. Serum was harvested at 48 hours of age, and FTPI was assessed using a digital Brix refractometer for total solids measurements, and digital refractometry and the biuret method to measure serum total protein (STP) concentrations. Serum gamma-glutamyltransferase activity was measured with an automated analyzer, and serum IgG was measured with the zinc sulfate turbidity test and an enzyme-linked immunosorbent assay. Diagnostic test performance was compared with that of the reference method (FTPI defined as a serum total IgG concentration <10 g/L). Test performance was evaluated using the area under the receiver operating characteristic curve, the sensitivity, the specificity, and the positive likelihood ratio at the optimal test cut point, and by calculating the kappa coefficient. RESULTS: A serum digital Brix percentage of <7.8% and an STP concentration of <52 g/L measured using digital refractometry were the best methods to identify calves with FTPI. The STP concentration measured with digital refractometry was 0.1 g/L lower than that measured with the biuret method. CONCLUSIONS: The digital Brix refractometer and the digital refractometer provide accurate and clinically useful methods for identifying dairy calves with FTPI. In this study, the excellent performance of the Brix refractometer was likely due to the use of a fixed sample volume (200 µL) and a uniform sample temperature at the time of measurement.

A Laboratory Diagnostic Approach to Hepatobiliary Disease in Small Animals.

Routine biochemical tests generally include serum enzymes, proteins, and other markers useful for identifying hepatobiliary disease in dogs and cats. Obtaining results outside the reference intervals can occur with direct hepatocellular injury, enzyme induction by hepatocytes or biliary epithelium, or decreased hepatic function. However, detection of biochemical abnormalities does not necessarily indicate clinically significant disease. For a comprehensive approach to detection and treatment of hepatobiliary disease, the laboratory results must be correlated with the history and physical examination findings, diagnostic imaging results, and other assays.

Clinical Approach to Advanced Renal Function Testing in Dogs and Cats.

Serum creatinine concentration is insensitive for detecting kidney injury and does not assist in differentiation between glomerular versus tubular damage. Advanced renal function tests, including glomerular filtration rate testing, determining fractional excretion of electrolytes, and assay of urine biomarkers, may allow earlier detection of reduced renal function mass, differentiation of renal from non-renal causes of azotemia, and assist with localization of damage. This article reviews the principles, indications, and limitations of these tests and describes their use in sample clinical scenarios.

Diagnosis of Small Intestinal Disorders in Dogs and Cats.

Laboratory tests are an important part of the workup of small intestinal diseases in dogs and cats. Especially in chronic cases, when extragastrointestinal causes need to be ruled out, it is important to adhere to a systematic workup. This article details the newest available data on tests to aid this diagnostic process. Once the diagnosis of a chronic enteropathy is made, there are many laboratory tests that can help in monitoring the disease and providing prognostic information. Several new tests being evaluated for clinical usefulness are discussed.

Practical Interpretation and Application of Exocrine Pancreatic Testing in Small Animals.

The pancreas remains a difficult organ to evaluate using laboratory methods alone. No single laboratory test is diagnostic of pancreatitis (chronic or acute) without other diagnostic modalities concurring with the diagnosis or ruling out other diseases. The diagnosis of pancreatitis is particularly difficult in cats, and pancreatitis often occurs with other diseases. The use of pancreatic cytology may be useful in diagnosing both inflammation and neoplasia. Exocrine pancreatic insufficiency (EPI) can be relatively easily diagnosed when clinically manifested by the measurement of trypsinlike immunoreactivity. Diagnosis is more difficult when EPI is subclinical.

ASVCP guidelines: quality assurance for point-of-care testing in veterinary medicine.

Point-of-care testing (POCT) refers to any laboratory testing performed outside the conventional reference laboratory and implies close proximity to patients. Instrumental POCT systems consist of small, handheld or benchtop analyzers. These have potential utility in many veterinary settings, including private clinics, academic veterinary medical centers, the community (eg, remote area veterinary medical teams), and for research applications in academia, government, and industry. Concern about the quality of veterinary in-clinic testing has been expressed in published veterinary literature; however, little guidance focusing on POCT is available. Recognizing this void, the ASVCP formed a subcommittee in 2009 charged with developing quality assurance (QA) guidelines for veterinary POCT. Guidelines were developed through literature review and a consensus process. Major recommendations include (1) taking a formalized approach to POCT within the facility, (2) use of written policies, standard operating procedures, forms, and logs, (3) operator training, including periodic assessment of skills, (4) assessment of instrument analytical performance and use of both statistical quality control and external quality assessment programs, (5) use of properly established or validated reference intervals, (6) and ensuring accurate patient results reporting. Where possible, given instrument analytical performance, use of a validated 13s control rule for interpretation of control data is recommended. These guidelines are aimed at veterinarians and veterinary technicians seeking to improve management of POCT in their clinical or research setting, and address QA of small chemistry and hematology instruments. These guidelines are not intended to be all-inclusive; rather, they provide a minimum standard for maintenance of POCT instruments in the veterinary setting.

[Reference values for haematological and clinical-chemical parameters in the dog]. / Hämatologische und klinisch-chemische Referenzwerte für Hunde.

OBJECTIVE: The aim of the study was to establish reference values for haematological and clinical-chemical parameters in adult dogs and to evaluate whether they are influenced by age, sex and feeding. MATERIAL AND METHODS: The study population consisted of 508 clinically healthy dogs of both genders and different breeds aged between ≤ 1 and 17 years. To establish reference values, results of 396 dogs aged 1-9 years were used. Analysis of haematological and clinical-chemical parameters was performed using the following devices: Cell-Dyn 3500, Hitachi 911, GEM Premier 3000 and the coagulometer BE CL 4. The statistical program SPSS was used to calculate the reference values. RESULTS: In 75% of the tested parameters no major discrepancies were noted in comparison to existing reference values. For eosinophils, basophils, monocytes, alanine aminotransferase, alkaline phosphatase, glutamate dehydrogenase, lipase, creatine kinase, bilirubin and creatinine cleardeviationsfromthe existing referencevalues were demonstrated. The reference ranges for eosinophils, monocytes and glutamate dehydrogenase were higher than reported inthe literature whereas for basophils they were lower. The reference ranges for the enzymes alanine aminotransferase, alkaline phosphatase, and lipase in young (< 1 year) and old dogs (≥ 10 years) differed significantly from the reference values of dogs aged 1-9 years. CONCLUSION AND CLINICAL RELEVANCE: Reference require regular re-evaluation because through continuing developments and new knowledge, some factors related to their determination, particularly methods and equipment, change.

Current quality assurance concepts and considerations for quality control of in-clinic biochemistry testing.

Quality assurance is an implied concept inherent in every consumer's purchase of a product or service. In laboratory testing, quality assurance encompasses preanalytic (sampling, transport, and handling prior to testing), analytic (measurement), and postanalytic (reporting and interpretation) factors. Quality-assurance programs require that procedures are in place to detect errors in all 3 components and that the procedures are characterized by both documentation and correction of errors. There are regulatory bodies that provide mandatory standards for and regulation of human medical laboratories. No such regulations exist for veterinary laboratory testing. The American Society for Veterinary Clinical Pathology (ASVCP) Quality Assurance and Laboratory Standards Committee was formed in 1996 in response to concerns of ASVCP members about quality assurance and quality control in laboratories performing veterinary testing. Guidelines for veterinary laboratory testing have been developed by the ASVCP. The purpose of this report was to provide an overview of selected quality-assurance concepts and to provide recommendations for quality control for in-clinic biochemistry testing in general veterinary practice.

Evidence of selective packaging and different α-granule subtypes in canine platelets.

Platelets are circulating megakaryocytic cytoplasmic fragments that are required to maintain vascular integrity and prevent haemorrhage. During activation, platelets release hundreds of bioactive proteins, mainly by secretion of α-granule content. These proteins include von Willebrand factor (vWF) and fibrinogen, major adhesive proteins involved in haemostasis. Human and mouse platelet α-granules are packaged selectively to contain different molecules and platelets can differentially release these proteins on specific receptor activation. Confocal laser scanning microscopy was used to analyse vWF and fibrinogen localization within canine platelet α-granules. In resting canine platelets, the majority of platelet vWF and fibrinogen is located in separate α-granule populations. These findings provide evidence that canine platelets contain different α-granule subtypes, suggesting that selective protein packaging occurs during canine platelet ontology.

ASVCP quality assurance guidelines: control of preanalytical, analytical, and postanalytical factors for urinalysis, cytology, and clinical chemistry in veterinary laboratories.

In December 2009, the American Society for Veterinary Clinical Pathology (ASVCP) Quality Assurance and Laboratory Standards committee published the updated and peer-reviewed ASVCP Quality Assurance Guidelines on the Society's website. These guidelines are intended for use by veterinary diagnostic laboratories and veterinary research laboratories that are not covered by the US Food and Drug Administration Good Laboratory Practice standards (Code of Federal Regulations Title 21, Chapter 58). The guidelines have been divided into 3 reports: (1) general analytical factors for veterinary laboratory performance and comparisons; (2) hematology, hemostasis, and crossmatching; and (3) clinical chemistry, cytology, and urinalysis. This particular report is one of 3 reports and documents recommendations for control of preanalytical, analytical, and postanalytical factors related to urinalysis, cytology, and clinical chemistry in veterinary laboratories and is adapted from sections 1.1 and 2.2 (clinical chemistry), 1.3 and 2.5 (urinalysis), 1.4 and 2.6 (cytology), and 3 (postanalytical factors important in veterinary clinical pathology) of these guidelines. These guidelines are not intended to be all-inclusive; rather, they provide minimal guidelines for quality assurance and quality control for veterinary laboratory testing and a basis for laboratories to assess their current practices, determine areas for improvement, and guide continuing professional development and education efforts.

Clinical chemistry parameters of piglets at weaning are modulated by an oral, low-dose interferon-alpha treatment.

Clinical chemistry parameters were investigated in piglets weaned at 22 and 28 days. The effects of an oral, low-dose interferon (IFN)-alpha treatment at weaning were evaluated as well. The trial was carried out on 59 piglets from the same farm, allocated to three groups: the first and the second groups were weaned at 28 and 22 days of age, respectively; the third group was weaned at 22 days and orally treated at weaning with IFN-alpha at a low dose (1 IU human lymphoblastoid IFN-alpha /kg body weight in drinking water) for 10 consecutive days. The results of the field trial confirmed that weaning is one of the main stressing events for pigs at intensive farms. In particular, these findings are based on a dramatic increase in serum haptoglobin levels after weaning in the three groups under study. Results also indicated that early weaning at 22 days implies higher environmental adaptation. In such animals, an oral, low-dose IFN-alpha treatment gave rise to a peculiar, negative, acute-phase response (reduced levels of serum albumin) and to significantly lower alpha-globulin concentrations in sera. Taken together, IFN-alpha was shown to modulate inflammatory responses to early weaning stress.

Technical note: Evaluation of a continuous ruminal pH measurement system for use in noncannulated small ruminants.

The objective of this study was to evaluate the precision and accuracy of an indwelling ruminal pH measurement system that could be used in small ruminants (small ruminant ruminal pH measurement system; SRS) without requiring ruminal cannulation. The outer diameter, length, and weight of the SRS were 20.6 mm, 138 mm, and 245 g, respectively. This device was capable of logging pH, temperature, and battery voltage. In Exp. 1, a ruminally cannulated sheep (94 kg) was infused with a 40% (wt/vol) glucose solution to supply 5 g of glucose/kg of BW into the rumen. Ruminal pH was recorded every 30 s simultaneously using a portable pH meter and the SRS. In Exp. 2, 30 noncannulated sheep (72 +/- 10 kg of BW) were orally administered with a 40% glucose solution as described above (5 g of glucose/kg of BW; n = 22) or an equivalent volume of water (12.5 mL/kg of BW; n = 8). Sheep were slaughtered 3 h after the oral drench, and immediately after slaughter ruminal pH readings were measured manually using a portable pH meter and were compared with measurements recorded by the SRS. In Exp. 1, the relationship between manual pH measurement using a portable pH meter and the SRS (226 data pairs) had a Pearson correlation coefficient and concordance correlation coefficient of 0.97 and 0.96, respectively. Furthermore, the scale shift and location shift observed in Exp. 1 were 1.28 and 0.00, respectively. The relationship between measurements conducted manually using a portable pH meter and the SRS in Exp. 2 had Pearson and concordance correlation coefficients of 0.96 and 0.95, respectively. The respective scale and location shifts for Exp. 2 were 1.16 and 0.04. These results indicate that the measurements obtained from SRS were in agreement with simultaneous measurements manually conducted using a portable pH meter, suggesting that the SRS can be used to measure ruminal pH in noncannulated small ruminants.

Cystic renal disease in the domestic ferret.

Cystic renal diseases in domestic ferrets are a common anecdotal finding but have received scant systematic assessment. We performed a 17-y, case-control retrospective analysis of the medical records of 97 ferrets housed at our institution between 1987 and 2004, to determine the prevalence and morphotypes of cystic renal diseases in this species. Histologic sections stained with hematoxylin and eosin, Masson trichrome, or periodic acid-Schiff were evaluated by a comparative pathologist, and statistical analysis of hematologic and serum chemistry values was correlated with morphologic diagnosis. Of the 97 available records, 43 were eliminated due to lack of accompanying tissues. Of the 54 remaining cases, 37 (69% prevalence) had documented renal cysts, and 14 of the 54 ferrets (26%) had primary polycystic disease consisting of either polycystic kidney disease affecting renal tubules or, more commonly, glomerulocystic kidney disease. Secondary polycystic lesions were identified in 11 ferrets (20%), and 12 ferrets (22%) exhibited focal or isolated tubular cysts only as an incidental necropsy finding. Ferrets with secondary renal cysts associated with other developmental anomalies, mesangial glomerulopathy, or end-stage kidney disease had hyperphosphatemia and elevated BUN in comparison with those with primary cystic disease and elevated BUN compared with those without renal lesions. Although reflecting institutional bias, these results implicate primary and secondary cystic renal diseases as highly prevalent and underreported in the domestic ferret. In addition to the clinical implications for ferrets as research subjects and pets, these findings suggest a potential value for ferrets as a model of human cystic renal diseases.

Clinicopathological presentation of cardiac disease in cattle and its impact on decision making.

The records of 116 cattle suffering from cardiac disease were examined retrospectively. On the basis of the results of postmortem examinations there were 52 cases of endocarditis, 39 of pericarditis and 25 congenital cardiac defects. The most useful clinical tool for differentiating between these conditions was auscultation of the heart. The cases of pericarditis were characterised by muffled heart sounds, and the cases of endocarditis and congenital cardiac defects were characterised by a cardiac murmur. Endocarditis could be differentiated from congenital cardiac defects by the presence of a jugular pulse, venous distension, oedema, a reduced appetite, pain and polyarthritis, whereas congenital defects were associated with conformational abnormalities. These two conditions could also be differentiated by differences in the plasma sodium concentration, the albumin:globulin ratio, red blood cell count, lymphocyte count and haematocrit. The ability to differentiate between these three groups of cardiac diseases can help the veterinary practitioner in deciding whether treatment, economic salvage (slaughter for human consumption) or disposal (slaughter not for human consumption) is likely to be the best option.

Complete blood count, clinical chemistry, and serology profile by using a single tube of whole blood from mice.

Clinical pathology is a valuable means for assessing specific organ pathology and a screening tool for general animal health. Routine clinical pathology evaluation in mice usually includes whole blood for a complete blood count (CBC) and a clinical biochemistry analysis. Acquisition and analysis of these samples can be problematic due to the small volumes of blood that can be obtained from a mouse. Typically, a complete blood count requires blood from a tube containing an anticoagulant, whereas a clinical biochemistry profile needs blood from a serum clot tube. Because of the small volume that can be obtained, splitting the blood from a single mouse into 2 different tubes may result in inadequate samples to perform the desired tests or introduce inaccuracies. We explored the feasibility of using a single lithium heparin tube for generation of a CBC, biochemistry profile, and serology profile. We also evaluated the consistency of CBC data, including the quality of a peripheral blood smear taken from a lithium heparin or EDTA tube after various storage times. We found that CBC, biochemistry, and serology profiles could be obtained more readily when blood samples were placed in a single lithium heparin tube than in 2 separate tubes. In addition, the quality of blood smears and CBC results from the lithium heparin tube were comparable (with few exceptions) to those from an EDTA tube after prolonged storage.

Castor bean (Ricinus communis) toxicosis in a sheep flock.

This paper describes clinical, laboratory and pathological findings of sheep, which is intoxicated with castor bean. The source of intoxication was a miscellaneous garden waste. Forty-five animals showed clinical toxicosis and 17 died. The clinical signs included weakness, salivation, profuse watery diarrhoea, dehydration, mydriasis, teeth grinding, hypothermia and recumbency. The most significant haematological and biochemical findings were a high haematocrit, high concentration of serum BUN, creatinine and phosphorus and high activity of serum CK and AST. Pathology revealed severe gastroenteritis, cardiac haemorrhage and necrosis, hepatic necrosis and acute tubular necrosis in kidneys. Treatment included symptomatic and supportive care with fluid therapy and cathartic administration.

Influence of ascorbic acid supplementation on the haematological and clinical biochemistry parameters of male rabbits exposed to aflatoxin B1.

This study was undertaken to evaluate the effectiveness of L-ascorbic acid (AA) in alleviating the toxicity of aflatoxin B1 (AFB1) in male New-Zealand white rabbits. Five rabbits (6 months of age and mean body weight 3.12 kg) per group were assigned to 1 of 6 treatment groups: 0 mg AA and 0 mg AFB1/kg BW (control); 20 mg AA/kg BW; 15 microg AFB1/kg BW; 15 microg AFB1 plus 20 mg AA/kg BW; 30 pg AFB1/kg BW; 30 pg AFB1 plus 20 mg AA/kg BW. Rabbits were orally administered their respective doses every other day for 9 weeks, followed by a 9-week recovery period where all drugs were withdrawn. Evaluations were made for hemato-biochemical parameters and enzymatic activities. Results showed that AFB1 significantly (p < 0.05) decreased hemoglobin (Hb), total erythrocytic count (TEC) and packed cell volume (PCV), in a dose-dependent manner, and these effects were continued during the recovery period. Ascorbic acid caused an increase in these parameters, and alleviated the negative effect of AFB1 during the treatment period. Additionally, serum concentrations of total protein, albumin and glucose were significantly (P < 0.05) declined by treatment with the high dose of aflatoxin and these effects were continued during the recovery period. Ascorbic acid caused non-significant increases in these parameters and alleviated the harmful effect of AFB1. On the other hand, aflatoxin treatment caused significant increases (P < 0.05) in the activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (AlP) during the treatment period in a dose dependent manner, and this effect was continued during the recovery period, especially with the high dose. Also, treatment with the high dose of aflatoxin caused significant increases (P<0.05) in cholesterol and total bilirubin. Ascorbic acid caused significant decreases in these parameters and alleviated the harmful effects of AFB1. Whereas, Total leukocyte count (TLC), urea and creatinine were not significantly affected by aflatoxin-treatment. Generally, it is interesting feature that the treatment with AA alone had no negative effects on most of the previous parameters. Also, the presence of AA could diminished the adverse effects of AFB1 on most of hematological and biochemical values, and enzymatic activities in rabbits.

Immunohistochemical study on hypoxia in spontaneous polycystic liver and kidney disease in rats.

Hypoxia-inducible factor (HIF) mediates homeostatic responses to hypoxia and activates transcription of hypoxia-inducible genes including vascular endothelial growth factor (VEGF). The aim of this study was to examine the expressions of VEGF, HIF-1alpha and HIF-3alpha in spontaneously occurring hepatorenal polycystic lesions in two Sprague-Dawley (Crj:CD) rats. Hepatic multiple cysts were derived from the interlobular and large bile ducts, while renal cysts were from the collecting ducts and distal tubuli. These findings were confirmed by a lectin peanut agglutinin (PNA) histochemistry. In the polycystic liver, VEGF immunoreaction was strongly evident in the cytoplasm of hepatocytes, whereas expression of HIF-3alpha, but not HIF-1alpha, was found in a few nuclei of hepatocytes. In the polycystic kidney, VEGF immunoreaction was increased in the cytoplasm of collecting ducts and distal tubuli, whereas nuclear expression of HIF-1alpha and HIF-3alpha was evident in the proximal tubuli and thin loop of Henle, respectively. The results suggest that hypoxia-related molecules may be induced by cystic alterations in a heterogeneous appearance.