Reference Value Advisor: a new freeware set of macroinstructions to calculate reference intervals with Microsoft Excel

Abstract: International recommendations for determination of reference intervals have been recently updated, especially for small reference sample groups, and use of the robust method and Box–Cox transformation is now recommended. Unfortunately, these methods are not included in most software programs used for data analysis by clinical laboratories. We have created a set of macroinstructions, named Reference Value Advisor, for use in Microsoft Excel to calculate reference limits applying different methods. For any series of data, Reference Value Advisor calculates reference limits (with 90% confidence intervals [CI]) using a nonparametric method when n≥40 and by parametric and robust methods from native and Box–Cox transformed values; tests normality of distributions using the Anderson–Darling test and outliers using Tukey and Dixon–Reed tests; displays the distribution of values in dot plots and histograms and constructs Q–Q plots for visual inspection of normality; and provides minimal guidelines in the form of comments based on international recommendations. The critical steps in determination of reference intervals are correct selection of as many reference individuals as possible and analysis of specimens in controlled preanalytical and analytical conditions. Computing tools cannot compensate for flaws in selection and size of the reference sample group and handling and analysis of samples. However, if those steps are performed properly, Reference Value Advisor, available as freeware at http://www.biostat.envt.fr/spip/spip.php?article63 , permits rapid assessment and comparison of results calculated using different methods, including currently unavailable methods. This allows for selection of the most appropriate method, especially as the program provides the CI of limits. It should be useful in veterinary clinical pathology when only small reference sample groups are available.     

Hematology and serum clinical chemistry reference intervals for free‐ranging Scandinavian gray wolves (Canis lupus)

Background: Scandinavian free‐ranging wolves (Canis lupus) are endangered, such that laboratory data to assess their health status is increasingly important. Although wolves have been studied for decades, most biological information comes from captive animals. Objectives: The objective of the present study was to establish reference intervals for 30 clinical chemical and 8 hematologic analytes in Scandinavian free‐ranging wolves. Methods: All wolves were tracked and chemically immobilized from a helicopter before examination and blood sampling in the winter of 7 consecutive years (1998–2004). Seventy‐nine blood samples were collected from 57 gray wolves, including 24 juveniles (24 samples), 17 adult females (25 samples), and 16 adult males (30 samples). Whole blood and serum samples were stored at refrigeration temperature for 1–3 days before hematologic analyses and for 1–5 days before serum biochemical analyses. Reference intervals were calculated as 95% confidence intervals except for juveniles where the minimum and maximum values were used. Results: Significant differences were observed between adult and juvenile wolves for RBC parameters, alkaline phosphatase and amylase activities, and total protein, albumin, γ‐globulins, cholesterol, creatinine, calcium, chloride, magnesium, phosphate, and sodium concentrations. Conclusion: Compared with published reference values for captive wolves, reference intervals for free‐ranging wolves reflected exercise activity associated with capture (higher creatine kinase activity, higher glucose concentration), and differences in nutritional status (higher urea concentration).     

Reference values: a review

Abstract: Reference values are used to describe the dispersion of variables in healthy individuals. They are usually reported as population‐based reference intervals (RIs) comprising 95% of the healthy population. International recommendations state the preferred method as a priori nonparametric determination from at least 120 reference individuals, but acceptable alternative methods include transference or validation from previously established RIs. The most critical steps in the determination of reference values are the selection of reference individuals based on extensively documented inclusion and exclusion criteria and the use of quality‐controlled analytical procedures. When only small numbers of values are available, RIs can be estimated by new methods, but reference limits thus obtained may be highly imprecise. These recommendations are a challenge in veterinary clinical pathology, especially when only small numbers of reference individuals are available.     

Determination of haematological reference intervals in healthy adult greyhounds

Objective : The purpose of this study was to establish breed‐specific reference intervals for haematological measurands in non‐racing greyhounds. Suitability of the data for partitioning according to sex was also examined. Methods : Haematological data were collected from 304 healthy non‐racing greyhounds and analysed using non‐parametric methods. Results were compared with non‐breed‐specific canine reference intervals and also with greyhound reference intervals obtained by other investigators. Results : Compared with non‐breed‐specific reference intervals, the results showed comparable mean and upper limit and higher lower limit for erythrocyte count; higher values for haemoglobin, haematocrit and mean corpuscular volume; and lower values for total leucocyte count and absolute concentration of neutrophils, lymphocytes, monocytes, eosinophils and platelets. Partitioning according to sex was recommended by the statistical analysis for all analytes except haematocrit and total leucocyte count. Clinical Significance : In this study the reference intervals were derived from a large sample size. The results are in general agreement with previous reports, although higher values for low reference limits have been noted for the erythroid parameters, and lower values for upper reference limits have been observed for the total and different leucocyte counts. Breed‐specific reference intervals provide a useful clinical tool for haematological evaluations.     

Determination of serum biochemistry reference intervals in a large sample of adult greyhounds

Background : As large breed, highly muscled dogs, greyhounds are regarded as physiologically different to other breeds. Biochemistry reference intervals have previously been determined using small numbers of greyhounds or based on the observations from racing dogs. Objectives : The purpose of this study was to develop statistically defined reference intervals for biochemical analytes in healthy non‐racing greyhounds. Partitioning according to gender was also examined. Methods : Biochemical analytes from a population of non‐racing healthy greyhounds, including 269 males, 202 females and 28 dogs where gender had not been recorded, were examined using a non‐parametric statistical approach. The dogs were aged between one and nine years old. Results : Total protein, albumin, globulin and creatinine reference intervals differed from generic reference intervals used for dogs. The reference intervals for greyhounds in this study are similar to those obtained by other studies of greyhounds, but often had a narrower range of values, likely reflecting more accurate estimation associated with larger numbers of reference individuals. Recommended methods for assessment of partitioning do not indicate a need to partition according to gender.     

Serum biochemical reference intervals for wild dwarf ornate wobbegong sharks (Orectolobus ornatus)

Background: Sharks are important to sport and commercial fishing, public aquaria, and research institutions. However, serum biochemical reference values have been established for few species. Objective: The aim of this study was to establish serum biochemical reference intervals for wild‐caught dwarf ornate wobbegong sharks (Orectolobus ornatus). Methods: Fifty wobbegongs were caught, and their health status, sex, length, and weight were evaluated and recorded. Following collection of blood, serum biochemical analytes were measured and analyzed using standard analytical and statistical methods. Combined samples generated means, medians, and reference intervals. Results: For the measured analytes, means (reference intervals) were as follows: sodium 287 (284–289) mmol/L, chloride 277 (274–280) mmol/L, potassium 5.2 (5.0–5.3) mmol/L, total calcium 4.6 (4.5–4.7) mmol/L, magnesium 1.9 (1.7–2.0) mmol/L, inorganic phosphate 1.8 (1.7–1.9) mmol/L, glucose 2.6 (2.4–2.8) mmol/L, total protein 46 (45–47) g/L, urea 396 (392–401) mmol/L, creatinine ≤0.02 mmol/L, total bilirubin 2.0 (1.9–2.1) μmol/L, cholesterol 1.3 (1.2–1.4) mmol/L, triglyceride 0.5 (0.4–0.6) mmol/L, alkaline phosphatase 24 (21–28) U/L, alanine aminotransferase 3 U/L, aspartate aminotransferase 28 (25–31) U/L, creatine kinase 49 (38–59) U/L, and osmolarity 1104 (1094–1114) mmol/L. Serum values were not affected by sex, length, or weight. Conclusions: Established reference values will assist with clinical evaluation and treatment of dwarf ornate wobbegongs in aquaria, research institutions, and the wild.     

Effect of age on reference intervals of serum biochemical values in kittens

OBJECTIVE: To determine the effect of age on reference intervals of serum biochemical values in kittens. DESIGN: Prospective clinical trial. ANIMALS: 55 kittens from 12 specific-pathogen-free queens. PROCEDURE: Kittens were allocated at birth into colostrum-fed (n = 27) and colostrum-deprived (28) groups. Blood was collected at birth and on days 1, 2, 4, 7, 14, 28, and 56. Serum samples were analyzed for activities of alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, creatine kinase, lactate dehydrogenase, gamma-glutamyltransferase, amylase, and lipase and for concentrations of albumin, total protein, bilirubin, urea nitrogen, creatinine, cholesterol, glucose, calcium, phosphorus, and triglycerides by use of an automated analyzer. Total serum solids concentrations were determined by use of refractometry. Serum IgG concentrations were quantified by use of radial immunodiffusion. RESULTS: For several analytes, reference intervals changed rapidly, most notably during the first few days of life. Reference intervals for alkaline phosphatase, creatine kinase, lactate dehydrogenase, and triglycerides were higher from birth to 8 weeks than adult reference intervals. Aspartate aminotransferase, bilirubin, urea nitrogen, and creatinine were higher than in adults at birth but were similar to or lower than adult reference intervals by 8 weeks. Compared with adult reference intervals, reference intervals for calcium and phosphorus concentrations were higher and for albumin and total protein concentrations were lower throughout the study period. CONCLUSIONS AND CLINICAL RELEVANCE: Important differences exist between reference intervals for serum biochemical values of neonatal and adult cats. Age-appropriate reference intervals should be used for accurate assessment of serum biochemical test results in cats.     

Hematologic and biochemical reference intervals for Norwegian crossbreed grower pigs

Background: Hematologic and biochemical reference intervals depend on many factors, including environment and age. Reference intervals for Norwegian grower pigs are lacking, and previously published reference intervals for similar pigs from other countries are now outdated due to significant changes in management and breeding on the pig farms. Objectives: The aim of this study was to determine updated reference intervals for hematologic and biochemical analytes in healthy crossbred grower pigs, and to compare the results among 3 different farms. Methods: Blood samples were collected from 104 clinically healthy pigs of the most common Norwegian crossbreed (Landrace Yorkshire sow × Landrace Duroc boar). The pigs were 12–16 weeks old, weighed 30–50 kg, of both sexes, and lived on 3 farms in eastern Norway. Automated hematologic and biochemical analysis were performed using ADVIA 2120 and ADVIA 1650 analyzers. Results: Five samples were excluded because of hemolysis (1) or outliers (4). Reference intervals were calculated using parametric or nonparametric methods, depending on data distribution. Mean, median, minimum, and maximum values were tabulated. Conclusions: The reference intervals calculated in this study will be useful for the diagnosis and monitoring of disease in this widespread crossbreed pig. Compared with previously published reference values, reference intervals for total WBC count, creatine kinase and alanine aminotransferase activities, and albumin, bilirubin, and urea concentrations in this study differed notably.     

Reference intervals for hematologic and biochemical constituents and protein electrophoretic fractions in captive common buzzards (Buteo buteo)

BACKGROUND: Increasing interest in wildlife care leads to the need for new tools to evaluate animal health. Laboratory investigations require reference intervals against which to compare the results obtained. For common buzzards, only a few studies have been performed to establish hematologic and biochemical reference intervals. OBJECTIVES: The aim of this work was to develop reference values for routine hematologic and biochemical constituents and protein electrophoretic fractions and evaluate possible seasonal differences in values for healthy common buzzards. METHODS: Heparinized blood samples were collected from 23 captive, clinically healthy common buzzards between February 2001 and June 2003. A CBC, routine biochemical analysis, and protein electrophoresis were performed. Data distribution was assessed and results from birds sampled in spring, summer, and winter were compared. Results from alternative methods for hemoglobin (Hgb; estimated as HCT / 3 vs spectrophotometry), total protein (biuret vs refractometry), and albumin (bromcresol green vs electrophoresis) concentrations also were compared. RESULTS: Reference intervals were calculated as 10-90th percentiles. In spring and summer, total WBC and heterophil counts, and urea, total protein, prealbumin, and beta- and gamma-globulins concentrations were significantly different from winter values. Results obtained by alternative methods for Hgb, total protein, and albumin concentrations were significantly different from those obtained by standard methods, although estimated and spectrophotometric Hgb values were significantly correlated. CONCLUSIONS: The reference values obtained in this study for hematologic and plasma biochemical constituents and their seasonal variation in healthy, captive common buzzards will be useful in the clinical evaluation of these birds in rehabilitation settings.     

ASVCP reference interval guidelines: determination of de novo reference intervals in veterinary species and other related topics

Reference intervals (RI) are an integral component of laboratory diagnostic testing and clinical decision‐making and represent estimated distributions of reference values (RV) from healthy populations of comparable individuals. Because decisions to pursue diagnoses or initiate treatment are often based on values falling outside RI, the collection and analysis of RV should be approached with diligence. This report is a condensation of the ASVCP 2011 consensus guidelines for determination of de novo RI in veterinary species, which mirror the 2008 Clinical Laboratory and Standards Institute (CLSI) recommendations, but with language and examples specific to veterinary species. Newer topics include robust methods for calculating RI from small sample sizes and procedures for outlier detection adapted to data quality. Because collecting sufficient reference samples is challenging, this document also provides recommendations for determining multicenter RI and for transference and validation of RI from other sources (eg, manufacturers). Advice for use and interpretation of subject‐based RI is included, as these RI are an alternative to population‐based RI when sample size or inter‐individual variation is high. Finally, generation of decision limits, which distinguish between populations according to a predefined query (eg, diseased or non‐diseased), is described. Adoption of these guidelines by the entire veterinary community will improve communication and dissemination of expected clinical laboratory values in a variety of animal species and will provide a template for publications on RI. This and other reports from the Quality Assurance and Laboratory Standards (QALS) committee are intended to promote quality laboratory practices in laboratories serving both clinical and research veterinarians.     

Genetic and phenotypic parameters for monthly egg production in White Leghorn hens

This article reports genetic and phenotypic parameters of monthly egg production and the influence of Box‐Cox transformation on the parameters from a population of White Leghorns, selected for feed efficiency. A total of 6450 daughters of 180 sires and 1335 dams were analysed by restricted maximum likelihood (REML) using a multivariate animal model. The traits considered were monthly egg productions, cumulative production of the first 5 months (S5), cumulative production of first 10 months (S10), and survivor egg production in the first cycle (S12). Two sets of data were analysed: the original data and with the Box‐Cox method transformed data. The results indicated that there were no great differences in the estimates between untransformed and transformed data. The estimates of heritability for monthly egg production were high for the first period, decreased to reach the lowest during peak production, and increased to the end of lay. The estimates of heritability for cumulative records were generally higher than monthly records. Genetic and phenotypic correlations among monthly egg production totals were generally high for contiguous periods and then decreased as the interval between months increased. The highest genetic correlation between monthly records and S5 was for the second month of production, whereas the correlations between monthly production totals and S10 and S12 reached their peak at the sixth and eighth months of production, respectively.     

The time factor in egg formation for hens exposed to ahemeral light-dark cycles

Three hundred and two White Leghorn pullets were killed at intervals 5 min to 7 h following oviposition. The reproductive tract was then examined and, from the position of the yolk, estimates were made of the mean time spent by the ovum in different parts of the oviduct. Half the birds examined had been kept under 24‐h cycles (14 h light : 10 h dark) and half under 27‐h cycles (14 h light : 13 h dark). Mean intervals from oviposition to ovulation were 24 min and 36 min for birds under 24‐h and 27‐h cycles respectively. The difference between these values can be associated with the different rates of lay of birds under the two treatments. Mean times spent in the upper oviduct and the shell gland were estimated as 5·15 h and 19·76 h for the 24‐h cycles and 5·41 h and 20–70 h for the 27‐h cycles. The increase in albumen weight which has been reported to follow from the use of 27‐h cycles can thus be associated with extra time spent in the magnum. The reported increase in shell weight of 10% is more than would be expected from a 5% increase in time spent in the shell gland.     

Efficient Monte Carlo algorithm for restricted maximum likelihood estimation of genetic parameters

Monte Carlo (MC) methods have been found useful in estimation of variance parameters for large data and complex models with many variance components (VC), with respect to both computer memory and computing time. A disadvantage has been a fluctuation in round‐to‐round values of estimates that makes the estimation of convergence challenging. Furthermore, with Newton‐type algorithms, the approximate Hessian matrix might have sufficient accuracy, but the inaccuracy in the gradient vector exaggerates the round‐to‐round fluctuation to intolerable. In this study, the reuse of the same random numbers within each MC sample was used to remove the MC fluctuation. Simulated data with six VC parameters were analysed by four different MC REML methods: expectation‐maximization (EM), Newton–Raphson (NR), average information (AI) and Broyden's method (BM). In addition, field data with 96 VC parameters were analysed by MC EM REML. In all the analyses with reused samples, the MC fluctuations disappeared, but the final estimates by the MC REML methods differed from the analytically calculated values more than expected especially when the number of MC samples was small. The difference depended on the random numbers generated, and based on repeated MC AI REML analyses, the VC estimates were on average non‐biased. The advantage of reusing MC samples is more apparent in the NR‐type algorithms. Smooth convergence opens the possibility to use the fast converging Newton‐type algorithms. However, a disadvantage from reusing MC samples is a possible “bias” in the estimates. To attain acceptable accuracy, sufficient number of MC samples need to be generated.     

Symmetric dimethylarginine values in koalas (Phascolarctos cinereus) based on oxalate nephrosis status

Oxalate nephrosis is a prevalent renal disease in koalas (Phascolarctos cinereus) of the Mount Lofty Ranges population in South Australia. The symmetric dimethylarginine (SDMA) assay is widely used in companion animals to diagnose renal disease, particularly in the early stages. This study aimed to determine: (1) reference intervals for SDMA in koalas and (2) SDMA values of koalas with oxalate nephrosis. Blood samples were collected from 41 Mount Lofty Ranges koalas euthanased on welfare grounds. Koalas were necropsied and, based on renal histopathology, were classified as unaffected (n = 22) or affected (n = 19) by oxalate nephrosis. Serum or plasma samples were analysed for creatinine, urea and SDMA and urine samples for urine specific gravity (USG). The reference interval for SDMA in unaffected koalas was 2.4–22.9 μg/dL. In koalas with oxalate nephrosis, SDMA was elevated in 74% of cases above the upper limit of the confidence interval. SDMA was elevated in three affected koalas with normal creatinine values. A positive correlation was found between SDMA and creatinine (R = 0.775, P < 0.001) and SDMA and urea (R = 0.580, P < 0.001) and a negative correlation between SDMA and USG (R = −0.495, P = 0.027). In conclusion, SDMA correlates well with other commonly used tests of renal function in koalas and should be included as part of the standard diagnostic process to increase the accuracy of oxalate nephrosis diagnosis in koalas.     

Comparison of bovine hematology reference intervals from 1957 to 2006

Background: The most commonly used bovine hematology reference intervals were published in 1965. We found the results from healthy cattle in 2001 differed from those in many ways. Discovery of the original laboratory book used to calculate the 1965 values gave us the opportunity to evaluate whether hematology values of healthy cattle have changed over time. Objective: The purpose of this study was to establish hematology reference intervals for Holstein cows, compare selected hematologic results with similar population data from 1957, and compare these reference intervals with those of other North American veterinary schools and published values. Methods: Reference intervals were developed in 2001 using clinically healthy, bovine leukemia virus‐negative, mid‐lactation Holstein cows. Selected parts of the hemograms and neutrophil:lymphocyte (N:L) ratio were compared with those from healthy, age‐matched Holstein cows evaluated in 1957. Bovine reference intervals were solicited from clinical pathology laboratories in North American veterinary colleges and analyzed for population characteristics and method of analysis. Results: Between 1957 and 2001, mean neutrophil counts increased significantly, whereas lymphocyte, monocyte, and eosinophil counts and hemoglobin concentration decreased significantly. Mean N:L ratio increased significantly to 1.17. Most surveyed laboratories were using the 1965 reference intervals. Two other institutions that had developed reference intervals after 2000 had results similar to ours. Conclusions: Continued use of older bovine hematology reference intervals could lead to misinterpretation of within‐reference neutrophil counts as neutrophilia and under‐recognition of neutropenia, eosinophilia, monocytosis, or lymphocytosis. Use of N:L>1 as evidence of inflammation should be discontinued or used with great caution.     

Plasma creatinine in dogs: intra- and inter-laboratory variation in 10 European veterinary laboratories

Background

There is substantial variation in reported reference intervals for canine plasma creatinine among veterinary laboratories, thereby influencing the clinical assessment of analytical results. The aims of the study was to determine the inter- and intra-laboratory variation in plasma creatinine among 10 veterinary laboratories, and to compare results from each laboratory with the upper limit of its reference interval.

Methods

Samples were collected from 10 healthy dogs, 10 dogs with expected intermediate plasma creatinine concentrations, and 10 dogs with azotemia. Overlap was observed for the first two groups. The 30 samples were divided into 3 batches and shipped in random order by postal delivery for plasma creatinine determination. Statistical testing was performed in accordance with ISO standard methodology.

Results

Inter- and intra-laboratory variation was clinically acceptable as plasma creatinine values for most samples were usually of the same magnitude. A few extreme outliers caused three laboratories to fail statistical testing for consistency. Laboratory sample means above or below the overall sample mean, did not unequivocally reflect high or low reference intervals in that laboratory.

Conclusions

In spite of close analytical results, further standardization among laboratories is warranted. The discrepant reference intervals seem to largely reflect different populations used in establishing the reference intervals, rather than analytical variation due to different laboratory methods.     

Circannual variation in plasma adrenocorticotropic hormone concentrations in the UK in normal horses and ponies, and those with pituitary pars intermedia dysfunction

Reasons for performing study: Pituitary pars intermedia dysfunction (PPID) is a common endocrinopathy, frequently diagnosed via plasma adrenocorticotropic hormone (ACTH) concentrations. Seasonal variation in plasma ACTH concentrations has been described in normal horses prompting caution in diagnosing PPID at certain times of the year. The aims of this study were to determine appropriate reference intervals for equine plasma ACTH throughout the year; and to examine the circannual variation of plasma ACTH concentrations in PPID cases. Hypothesis: Plasma ACTH can be used as a test for PPID throughout the year with the use of appropriate reference intervals. Methods: Data for reference interval calculations were obtained from samples collected from inpatients of Liphook Equine Hospital (non‐PPID group, n = 156). Data from PPID cases (n = 941) were obtained from samples submitted to the Liphook Equine Hospital Laboratory from horses with a clinical suspicion of PPID found to have plasma ACTH concentrations greater than our upper reference interval for that time of year. Results: Upper limits for reference interval of plasma ACTH were 29 pg/ml between November and July and 47 pg/ml between August and October. Circannual variation in plasma ACTH occurred in both non‐PPID and PPID horses with the highest ACTH concentrations found between August and October in both groups (P<0.0001). The greatest difference between the 2 populations also occurred between August and October. Conclusions: Plasma ACTH can be used for the diagnosis and monitoring of PPID throughout the year with the use of appropriate reference intervals. These findings demonstrate an increase in pituitary gland secretory activity during the late summer and autumn in both normal and PPID cases.     

Haematology and plasma biochemistry of Stamboek pre-pubertal gilts in Italy: reference values

Blood samples were taken between February and April from 105 healthy Stamboek pre-pubertal gilts, aged 1-3 months, which were housed at a modern pig farm in northern Italy. The blood was analysed for nine haematological and nine selected haematochemical variables by means of automated and semiautomated blood analysis apparatus. After detection and rejection of outliers, the data were submitted to reference limits evaluation, also taking into account the limits for the red blood cell volume histogram as the anisocytosis index. Some haematological reference values deal with previously published data; in the haematochemical parameters, several discrepancies between evaluated limits and existing reference limits were noted, mainly for aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase levels and total protein concentration. The results confirm the relevance of age in determining blood reference intervals and that 'normal' values should be determined by each laboratory, taking into account the age of subjects, the sample size and methods of analysis.     

Determination of reference intervals for plasma biochemical values in clinically normal adult domestic shorthair cats by use of a dry-slide biochemical analyzer

OBJECTIVE: To establish reference intervals of plasma biochemical values in healthy adult domestic shorthair (DSH) cats by use of controlled conditions. ANIMALS: 95 healthy client-owned cats. PROCEDURES: Food was withheld from the cats overnight. All blood samples were obtained on the same day, at the same location, and by the same investigator. Blood samples were collected from a cephalic vein into lithium heparin tubes. After centrifugation of blood samples, plasma supernatants were harvested and stored at -20 degrees C until assayed for total proteins, albumin, creatinine, urea, glucose, calcium, phosphates, sodium, chloride, potassium, and CO2 concentrations and alkaline phosphatase and alanine aminotransferase activities. RESULTS: Reference intervals in healthy adult DSH cats were 65 to 85 g/L for total proteins, 27 to 39 g/L for albumin, 89 to 207 micromol/L for creatinine, 6.6 to 11.3 mmol/L for urea, 4.1 to 8.2 mmol/L for glucose, 2.4 to 2.9 mmol/L for calcium, 1.1 to 2.1 mmol/L for phosphates, 153 to 161 mmol/L for sodium, 120 to 127 mmol/L for chloride, 3.3 to 4.2 mmol/L for potassium, 15 to 21 mmol/L for CO2, 32 to 147 U/L for alkaline phosphatase, and 34 to 123 U/L for alanine aminotransferase. CONCLUSIONS AND CLINICAL RELEVANCE: This study provided reference intervals for plasma analytes in adult DSH cats. The influence of potential confounding factors was minimized through use of controlled preanalytic and analytic conditions. However, these results cannot be extrapolated to other feline breeds or used to interpret results from other biochemical analyzers.     

One-stage prothrombin time, activated partial thromboplastin time, thrombin time, fibrin degradation product concentration, and antithrombin activity in healthy adult alpacas

Background: Alpacas are increasingly presented to veterinarians for evaluation and care. Reports of alpaca reference intervals for one‐stage prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT), concentration of fibrin degradation products (FDP), and antithrombin (AT) activities are scarce or nonexistent. Objective: The aim of this study was to determine values for blood coagulation times (PT, aPTT, and TT), FDP concentrations, and AT activities in healthy adult alpacas. Methods: Of blood samples collected from 35 clinically healthy adult alpacas via jugular venipuncture and placed into sodium citrate and FDP tubes, 29 samples were assayable for coagulation testing. PT, aPTT, and TT were determined by physical (mechanical) clot detection; AT activity was determined using a thrombin‐specific chromogenic substrate end‐point assay; and FDP concentrations were determined by the slide agglutination method. Results: Median values and ranges (minimum–maximum) were determined for PT (8.7 seconds, 6.6–11.2 seconds), aPTT (17.3 seconds, 11.9–22.5 seconds), TT (10.2 seconds, 5.4–16.0 seconds), and AT activity (123.3%, 104.8–144.2%). The mean concentration of FDP was <8 μg/mL. Conclusion: These values for coagulation times, FDP concentration, and AT activity will provide a useful starting point in the diagnostic evaluation of ill adult alpacas.