Evaluation of serum enzyme activities as predictors of passive transfer status in lambs

OBJECTIVE: To determine the associations between serum IgG concentration and serum activities of gamma-glutamyltransferase (GGT), alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and pseudocholinesterase for the potential use of these serum enzymes as predictors of passive transfer status in neonatal lambs. DESIGN: Prospective observational study. ANIMALS: 47 Sardinian lambs from birth to 2 days old. PROCEDURE: Serum enzyme activities were measured by use of commercially available kits and a clinical biochemical analyzer. Serum IgG concentration was determined by single radial immunodiffusion. Associations between serum IgG concentration and the activity of each serum enzyme were established by use of regression analysis. RESULTS: A significant correlation was detected between serum IgG concentration and serum GGT activity in 1- and 2-day-old lambs. Minimal correlations were detected between serum IgG concentration and serum alkaline phosphatase activity in 1-day-old lambs and serum pseudocholinesterase activity in 1- and 2-day-old lambs. No significant associations were detected between serum IgG concentration and serum activities of aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase. A multiple linear regression model was accurate for the estimation of the natural logarithm of serum IgG concentration as a function of the natural logarithm of serum GGT activity and of the age of lambs at the time of sampling (adjusted R2 = 0.89). This model was then used to calculate the serum GGT activity equivalent to various serum IgG concentrations for 1- and 2-day-old lambs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that passive transfer status in neonatal lambs can be successfully predicted by measurement of serum GGT activity but not by measurement of the other enzymes tested.     

Evaluation of assays for determination of passive transfer status in neonatal llamas and alpacas

OBJECTIVE: To evaluate several practice-adapted assays for determination of passive transfer status in crias. ANIMALS: 24 llama and 9 alpaca crias. DESIGN: Prospective study. PROCEDURE: Serum IgG concentration was measured by use of a radial immunodiffusion assay when crias were 45 to 51 hours old. Results were compared with serum gamma-glutamyltransferase (GGT) activity, serum total protein, albumin, globulin, and total solids concentrations, and results of commercially available and traditional sodium sulfite turbidity (SST) tests. RESULTS: Mean (+/- SD) serum IgG concentration was 1,762 +/- 1,153 mg/dl. On the basis of a threshold value of 1,000 mg of IgG/dl at 48 hours of age, 5 of 33 (15.15%) crias had failure of passive transfer. Serum total solids, protein, and globulin concentrations were significantly associated with serum IgG concentration, whereas serum GGT activity and serum albumin concentration were not. Serum IgG concentrations were significantly different among crias with negative, 2+, and 3+ scores on the traditional SST test. Serum IgG concentrations were not significantly different between crias with negative and 100 mg/dl scores or 100 and 300 mg/dl scores on the commercially available SST test. However, all other comparisons between crias with different scores revealed significant differences. Sensitivity and specificity ranged between 0 and 1, depending on the test and endpoint selected. CONCLUSIONS AND CLINICAL RELEVANCE: The commercially available SST test and determination of serum total protein and globulin concentrations are suitable methods for assessing passive transfer status in llama and alpaca crias.     

Use of refractometry for determination of psittacine plasma protein concentration

Background: Previous studies have demonstrated both poor and good correlation of total protein concentrations in various avian species using refractometry and biuret methodologies. Objectives: The purpose of the current study was to compare these 2 techniques of total protein determination using plasma samples from several psittacine species and to determine the effect of cholesterol and other solutes on refractometry results. Methods: Total protein concentration in heparinized plasma samples without visible lipemia was analyzed by refractometry and an automated biuret method on a dry reagent analyzer (Ortho 250). Cholesterol, glucose, and uric acid concentrations were measured using the same analyzer. Results were compared using Deming regression analysis, Bland–Altman bias plots, and Spearman's rank correlation. Results: Correlation coefficients (r) for total protein results by refractometry and biuret methods were 0.49 in African grey parrots (n=28), 0.77 in Amazon parrots (20), 0.57 in cockatiels (20), 0.73 in cockatoos (36), 0.86 in conures (20), and 0.93 in macaws (38) (P≤.01). Cholesterol concentration, but not glucose or uric acid concentrations, was significantly correlated with total protein concentration obtained by refractometry in Amazon parrots, conures, and macaws (n=25 each, P<.05), and trended towards significance in African grey parrots and cockatoos (P=.06). Conclusions: Refractometry can be used to accurately measure total protein concentration in nonlipemic plasma samples from some psittacine species. Method and species‐specific reference intervals should be used in the interpretation of total protein values.     

Comparison of four methods for determination of total protein concentrations in pleural and peritoneal fluid from dogs

OBJECTIVE: To compare 4 techniques for determination of total protein concentrations in peritoneal and pleural effusions from dogs. SAMPLE POPULATION: 23 peritoneal and 12 pleural fluid samples from 35 dogs with various abnormalities. PROCEDURE: Samples were collected into tubes containing EDTA, centrifuged, and stored at -20 C until total protein concentrations were assessed. Protein concentration in each sample was determined by use of urine test strips, refractometry, and Bradford and biuret techniques. Accuracy of each method was determined, using dilutions of human control sera. RESULTS: There was good correlation among results of all quantitative procedures. Results of the biuret technique were more accurate than results of the Bradford assay. Refractometry underestimated protein concentration in samples with < 20 g of protein/L. Results of urine test strips correctly classified effusion samples into 2 groups on the basis of total protein concentrations less than or greater than 20 g/L. CONCLUSIONS AND CLINICAL RELEVANCE: Results of any of these 4 techniques can be used to rapidly and efficiently differentiate peritoneal and pleural fluid from dogs into transudates and exudates on the basis of total protein concentration less than or greater than 20 g/L, respectively.     

Measurement of neonatal equine immunoglobulins for assessment of colostral immunoglobulin transfer: comparison of single radial immunodiffusion with the zinc sulfate turbidity test, serum electrophoresis, refractometry for total serum protein, and the sodium sulfite precipitation test.

Four procedures for assessment of adequacy of colostral immunoglobulin (Ig) transfer in foals were evaluated. Results of zinc sulfate turbidity test, serum electrophoresis, total serum protein refractometry, and sodium sulfite precipitation test were compared with immunoglobulin G content determined by single radial immunodiffusion. The zinc sulfate turbidity test gave acceptable results for IgG, except that hemolyzed serum samples gave higher than expected values. A correction factor for hemolyzed serum was found to be useful. Serum electrophoresis was a satisfactory method of estimating IgG content. Total serum protein values may not be a valid basis for estimating IgG content, inasmuch as postsuckling total protein values were found to decrease in some foals in which passive transfer of IgG had been adequate. Sodium sulfite precipitation reactions were too unpredictable to be of value for determination of neonatal IgG concentration.     

Detection of low serum immunoglobulin concentrations in clinically ill calves

The sensitivity, specificity, and accuracy of classification of 4 tests for failure of passive transfer (FPT) were examined in clinically ill neonatal calves. Comparisons were made with serum IgG1 concentrations determined by radial immunodiffusion. Serum samples were obtained from 27 clinically ill calves < or = 21 days of age. The results of 4 commonly used assays, the sodium sulfite turbidity test, the zinc sulfate turbidity test, refractometry, and the serum gamma-glutamyl transferase (GGT) activity test, were compared with radial immunodiffusion determinations of serum IgG1 concentration. Serum GGT activity using a 50 IU/L threshold resulted in correct classification of the highest percentage of calves (93%) with regard to their passive transfer status. The sodium sulfite test with a 1+ end point and refractometry using a 5.5 g/dL end point resulted in correct classification of 85% of the calves studied. When using the sodium sulfite test, the 2+ and 3+ test end points had lower specificity, 0.58 and 0.00, respectively, than the 1+ end point. This loss in specificity resulted in misclassification of calves with adequate serum immunoglobulin concentrations as having FPT. The zinc sulfate turbidity test was inadequately specific (0.33) and resulted in misclassification of 33% of calves.     

Effect of passive transfer status on preweaning growth performance in dairy lambs

OBJECTIVE: To evaluate the effect of passive transfer status, determined by measuring serum IgG concentration 24 hours after parturition, on preweaning growth performance in dairy lambs. DESIGN: Prospective observational study. ANIMALS: 20 healthy Sardinian dairy lambs. PROCEDURES: Serum IgG concentration was measured 24 hours after birth. Body weight was measured at birth and at the time of weaning 28 days (ie, 27 to 29 days) after birth. Mean daily gain from birth to day 28 and day 28 weight were used as measures of preweaning growth performance. Regression analysis was used to evaluate associations between serum IgG concentration 24 hours after birth and measures of preweaning growth performance. RESULTS: Mean +/- SD serum IgG concentration 24 hours after birth was 24.6 +/- 17.5 mg/mL. Mean body weights at birth and weaning were 2,696 +/- 937 g and 9,253 +/- 2,116 g, respectively, and mean daily gain was 234 +/- 63 g/d. No significant association was detected between serum IgG concentration 24 hours after birth and birth weight. However, serum IgG concentration 24 hours after birth was significantly associated with mean daily gain (R(2) = 0.25). Each 1 mg/mL increase in serum IgG concentration 24 hours after birth was associated with a 1.8 g/d increase in mean daily gain and a 60.8-g increase in day 28 weight. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that passive transfer status, determined as serum IgG concentration 24 hours after birth, was a significant source of variation in preweaning growth performance in dairy lambs.     

Changes in serum protein values of sheep infected with Fasciola gigantica.

Serum protein values were determined in 6 lambs given 1 dose of 200 metacercariae of Fasciola gigantica. Over the 22 weeks of the experiment, total serum protein values in these lambs were only moderately increased over those of control lambs (n = 6). The increase was mainly associated with the change in alpha2-globulin and gamma-globulin fractions. The highest value of gamma-globulin (39.03%) of total serum protein was observed at 18 weeks after infection. Mean albumin concentration in the exposed lambs decreased considerably, starting in week 8 and persisting until the end of the experiment. The lowest level (39.28%) was seen at week 16.     

Blood and colostrum/milk serum gamma-glutamyltransferase activity as a predictor of passive transfer status in lambs

The importance of blood and colostrum/milk serum gamma-glutamyl transferase (gamma-GT) enzyme activity was evaluated to assess passive transfer status in healthy lambs. Thirty Akkaraman sheep (3-6 years old) were used which had normal pregnancy period and the same conditions, and the age of the lambs ranged between 0 and 15 days. Blood and colostrum/milk samples were collected from sheep and lambs after birth, before suckling (0) and after on 1st, 3rd, 7th and 15th days. Serum immunoglobulin G (IgG) concentration was determined by the use of Single Radial Immunodiffusion method. Serum gamma-GT activity was measured, using a commercially available kit in blood and colostrum/milk samples. Correlations were carried out between immunoglobulin and gamma-GT levels. Regression models (simple and multiple) were calculated with significant data. Linear correlation was determined between colostrum/milk gamma-GT activity and IgG concentrations and between serum gamma-GT activity and IgG concentrations in lambs on the 0 day. (r: 0.607, P: 0.001), 1st (r: 0.768, P: 0.001) and the 3rd (r: 0.603, P: 0.001) days and on the 1st (r: 0.637, P: 0.001) and 3rd (r: 0.478, P: 0.012) days in the experiment, respectively. Multivariate regression models were developed to estimate sample IgG concentration. Serum and colostrum/milk IgG concentration could be predicted using the formula: lamb serum IgG = 825 + 0.688 (lamb gamma-GT) + 52 (days); colostrum/milk IgG = 832 + 0.505 (colostrum/milk gamma-GT) - 167 (days). The regression models were moderately accurate in predicting serum IgG concentration (R2 = 0.51) and colostrum/milk IgG concentration (R2 = 0.55). Test sensitivity and positive predictive values for serum gamma-GT enzyme activity were found to be 96 and 100% and for colostrum/milk gamma-GT enzyme activity were found to be 100 and 68% to prediction IgG concentration. Serum and colostrum/milk gamma-GT activity can be used to assess passive transfer status of lambs. Along with this, regression models used to calculate serum and colostrum/milk gamma-GT activities found to be useful to estimate sample IgG concentration. The use of serum and colostrum/milk gamma-GT enzyme activity was found useful especially after birth on the 0, 1st and 3rd days.     

Evaluation of a commercially available immunoassay for assessing adequacy of passive transfer in calves

OBJECTIVE: To evaluate diagnostic utility of a commercially available immunoassay for assessing adequacy of passive transfer of immunity in neonatal calves. DESIGN: Prospective study. ANIMALS: 123 calves. PROCEDURE: Blood and serum samples were obtained from the calves prior to 2 weeks of age. The immunoassay was performed, along with refractometry and an 18% sodium sulfite turbidity test. Serum IgG concentration was determined with a radial immunodiffusion assay. Sensitivity and specificity of the immunoassay, refractometry, and the sodium sulfite test were calculated by comparing results with results of the radial immunodiffusion assay. RESULTS: Sensitivity and specificity of the blood IgG immunoassay were 0.93 and 0.88, respectively, compared with 1.00 and 0.53 for the sodium sulfite test. For refractometry, sensitivity and specificity were 0.71 and 0.83, respectively, when a serum total solids concentration of 5.2 g/dl was used as the cutoff between positive and negative test results. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the immunoassay performs well in detecting calves with inadequate passive transfer of immunity.     

Evaluation of 3 Assays for Failure of Passive Transfer in Calves

This study examined the sensitivity, specificity, predictive values, and classification accuracy of 3 commonly used screening tests for failure of passive transfer: the sodium sulfite turbidity test, the zinc sulfate turbidity test, and re-fractometry relative to serum immunoglobulin G₁, (IgG₁) concentrations determined by radial immunodiffusion. Serum samples were obtained from 242 calves ranging from 1 to 8 days of age. Using a serum concentration of 1,000 mg/dL IgG₁ to define adequate passive transfer, the zinc sulfate test had a sensitivity of 1.00 and a specificity of 0.52 in the detection of inadequate passive transfer. The endpoint of the test appeared to be higher than desired; calves testing negative had mean serum IgG₁ concentration of 955 mg/dL and a large proportion of calves with adequate passive transfer were misclassified as positive for failure of passive transfer. Using the qualitative zinc sulfate test, the percentage of calves correctly classified with regard to passive transfer status was less than that observed with either the sodium sulfite test or refractometry. The sensitivity of the sodium sulfite assay was 0.85 at a 1+ endpoint and 1.00 at a 2 or 3+ endpoint. The specificity of the sodium sulfite assay varied from 0.87 at a 1+ endpoint and 0.56 at a 2+ endpoint. The sensitivity and specificity of refractometry varied from 0.01 to 1.00 depending on the choice of endpoint. Refractometry correctly classified the largest proportion of calves with regard to their passive transfer status at test endpoints of 5.0 and 5.5 g/dL, 83% and 82% respectively. The highest percentages of calves correctly classified occurred with the sodium sulfite test using a 1+ endpoint (86.30%) and refractometry using a 5.0 g/dL endpoint (83.00%). A regression equation was developed that permitted calculation of an optimal endpoint for refractometric determinations of total serum protein concentration. A serum protein concentration of 5.2 g/dL was equivalent to 1,000 mg/dL serum IgG₁. Optimal selection of tests for passive transfer status in calves will be governed by the prevalence of failure of passive transfer, test performance, and the anticipated costs of classification errors.     

Evaluation of surrogate markers for passive transfer of immunity in kittens

OBJECTIVE: To identify surrogate markers of passive transfer of immunity 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 (ALP), 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 estimated by use of refractometry. Serum IgG concentrations were quantified by use of radial immunodiffusion. RESULTS: All kittens were agammaglobulinemic at birth. Colostrum-fed kittens had significantly higher IgG concentrations than did colostrum-deprived kittens from 1 though 28 days of age. Transient significant differences in serum biochemical variables between the colostrum-deprived and colostrum-fed groups were substantially resolved by day 4. Passive transfer of immunity could be reliably determined at 1 day of age and to a lesser extent at 2 days of age only by measurement of serum activity of ALP. CONCLUSIONS AND CLINICAL RELEVANCE: Adequacy of passive transfer in kittens initially correlated with serum activity of ALP, but quantification of serum IgG concentration was necessary after 2 days of age.     

The usefulness and limitations of hand-held refractometers in veterinary laboratory medicine: an historical and technical review

Abstract: Medical hand-held refractometers have been used in veterinary practice since their development in the 1960s. They have become ubiquitous for the measurement of protein and urine solute concentrations because of their rapidity of analysis, ease of use, and relatively low cost. Refraction of light offers advantages for the determination of solute concentrations because the measurement requires no chemical alteration of the specimen. Numerous authors have reported that the results of protein estimation by refractometry for domestic mammals correlate well with those obtained by the biuret method, although others have reported both higher and lower refractometric results compared with biuret results. Major discrepancies between biuret and refractometric results have been reported for avian samples. Some of the variation in reported results may be due to differences in design by refractometer manufacturers. Another possible source may be variation in the biuret reagent mixture and assay conditions. Refractometers also can be used to calculate serum water concentration. A table that converts index of refraction to serum water concentration can be used to convert electrolyte concentration from mmol/L of serum to mmol/L of serum water, a more accurate indicator of effective electrolyte concentration. Refractometers are especially useful for determining urine specific gravity on veterinary samples because they require relatively small sample volumes. Specific gravity continues to be the most common unit for reporting total solids concentration. Some solutes, such as acetone, may cause false increases in specific gravity by refractometry, as they increase refraction but are less dense than water.     

Prediction of serum IgG1 concentration in beef calves based on age and serum gamma-glutamyl-transferase activity

This study examined the relationship between serum gamma-glutamyl-transferase (GGT) activity and passive transfer status in beef calves less than 18 days of age. Immunoglobulin G1 (IgG1) concentrations were measured in 69 commercial beef calves between the ages of 24 and 72 hours. GGT activities were then measured in these same calves at various ages between 3 and 18 days of age. Models were developed predicting serum IgG1 concentration as a function of calf age and serum GGT activity. Minimal association was present between initial serum IgG1 concentration and serum GGT activity when all calves less than 18 days of age were considered (r2 = 0.065). When the study population was restricted to calves less than 8 days of age, the fit of the developed model was greatly improved (r2 = 0.438). Serum GGT activity has no apparent advantage relative to other assay procedures for predicting passive transfer status in beef calves. If serum GGT activity is to be used to assess passive transfer status in beef calves, application of this procedure should be restricted to calves less than 8 days of age.     

Evaluation of five commercially available assays and measurement of serum total protein concentration via refractometry for the diagnosis of failure of passive transfer of immunity in foals

OBJECTIVE: To determine and compare sensitivity, specificity, accuracy, and predictive values of measurement of serum total protein concentration by refractometry as well as 5 commercially available kits for the diagnosis of failure of passive transfer (FPT) of immunity in foals. DESIGN: Prospective study. ANIMALS: 65 foals with various medical problems and 35 clinically normal foals. PROCEDURE: IgG concentration in serum was assessed by use of zinc sulfate turbidity (assay C), glutaraldehyde coagulation (assay D), 2 semiquantitative immunoassays (assays F and G), and a quantitative immunoassay (assay H). Serum total protein concentration was assessed by refractometry. Radial immunodiffusion (assays A and B) was used as the reference method. RESULTS: For detection of IgG < 400 mg/dL, sensitivity of assay H (100%) was not significantly different from that of assays C, E, and G (88.9%). Specificity of assays H (96.0%) and G (95.8%) was significantly higher than that of assays C (79.4%) and E (78.1 %). For detection of IgG < 800 mg/dL, sensitivities of assays H (976%), D (92.9%), C (81.0%), and G (81.0%) were significantly higher than that of assay F (52.4%). Specificity of assays F (100%), G (94.7%), and H (82.8%) was significantly higher than that of assays C (56.9%) and D (58.6%). Serum total protein concentration < or = 4.5 g/dL was suggestive of FPT, whereas values > or = 6.0 g/dL indicated adequate IgG concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: Most assays were adequate as initial screening tests. However, their use as a definitive test would result in unnecessary treatment of foals with adequate IgG concentrations.     

Prediction of Serum IgG1 Concentration in Holstein Calves Using Serum Gamma Glutamyltransferase Activity

We examined the relationship between serum gamma glutamyltransferase (GGT) activity and serum gamma immunoglobulin G (IgG1) concentration in Holstein calves. Blood samples were collected from calves aged 1 to 3 days. A follow-up sample was obtained from each calf 2, 7, or 15 days after the initial sampling. Serum GGT activity and lgG1 concentration were measured. Regression models were used to predict lgG1 concentration as a function of age and serum GGT activity measured 2, 7, or 15 days later.
Serum GGT activity and calf age at the time of the second sample were directly related to serum IgG1 concentration in the initial sample in calves aged 3 to 17 days ( r = .54) and in calves aged 3 to 10 days ( r = .63). Models were used to estimate the serum GGT activity equivalent to a serum IgG1 concentration of 1,000 mg/dL. One-day-old calves should have serum GGT activities > 200 IU/L. Four-day-old calves should have serum GGT activities > 100 IU/L. One-week-old calves should have serum GGT activities > 75 IU/L. Calves with serum GGT activities < 50 IU/L should be classified as having failure of passive transfer.     

Relationship of protein concentration and water content of equine serum and plasma samples

A highly significant correlation between the water content and protein concentration of equine serum and plasma samples was demonstrated over a wide range of concentrations. A close correlation was also observed between protein concentration as estimated by refractometry and as determined by the biuret procedure for equine serum and plasma samples.     

Determination of chicken and turkey plasma and serum protein concentrations by refractometry and the biuret method

Plasma and serum protein concentrations were determined in chickens and turkeys by refractometry (with human and veterinary refractometers) and by the biuret method. Chicken and turkey serum protein values were significantly lower than respective plasma protein values according to both methods. Refractometer readings for both plasma and serum correlated closely with the results of the biuret test (r2 = 0.72 to 0.97). These findings indicate that plasma and serum protein values may be determined accurately in chickens and turkeys with a handheld refractometer.     

Evaluation of a lateral-flow immunoassay for use in monitoring passive transfer of immunoglobulins in calves

OBJECTIVE: To determine the predictive ability of a commercially available lateral-flow immunoassay used for determining passive transfer of immunoglobulins in calves. ANIMALS: 204 male Holstein calves ranging from 4 to 8 days old. PROCEDURE: Serum samples were obtained from each calf. Results of refractometry, zinc sulfate turbidity technique, and the lateral-flow immunoassay were determined. Sensitivity, specificity, accuracy, and predictive ability were calculated on the basis of IgG concentrations determined by turbidimetric immunoassay (TIA). RESULTS: Mean IgG concentration in the study was 10.9 mg/ml as determined by TIA. Rate of failure of passive transfer in this study population was 56%. Associations between the values for the refractometry and zinc sulfate turbidity techniques were established by regression analysis. Accuracy for the lateral-flow immunoassay, refractometry, and zinc sulfate turbidity methods was 95, 80, and 73%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The lateral-flow immunoassay was better at determining the status of passive transfer of immunoglobulins, compared with the refractometry or zinc sulfate turbidity methods. The ability of the lateral-flow immunoassay to provide accurate results should enable clinicians to make immediate management or intervention decisions.     

Comparison of refractometer and biuret methods for total protein measurement in body cavity fluids

Abstract: Most hand-held medical refractometers have internal scales that limit protein measurement to results ≥ 2.5 g/dL. Tables for conversion of refraction (r) to protein concentration for values as low as 0.1 g/dL were published in the 1960s, but their accuracy for use on body fluids has not been established. The purpose of this study was to assess the reliability of body cavity fluid protein determination by refractometry. We compared the protein concentration of 25 body cavity fluids as determined by 2 Goldberg type hand-held refractometers with results obtained by the biuret method. Published charts converting refraction (r) to protein concentration were used to determine protein concentration in samples with protein <2.5 g/dL. Higher protein values were read directly from the instruments. The range of comparison was limited to ≥ 0.6 g/dL, the lowest concentration of the biuret method's standard curve. Twenty-one peritoneal fluid, 2 pleural fluid and 2 pericardial fluid samples from 16 horses, 5 cattle, 3 dogs, 2 llamas and 1 cat were tested. The results obtained by the two refractometers were closely and linearly related to biuret results (P<.001), with slopes by linear regression analysis close to 1, and correlation coefficients >0.977. Based on this study, the range for quantification of body cavity fluid protein concentration by refractometry can be extended below 2.5 g/dL, allowing for quantitative assessment of most clinical samples.