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.     

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.     

Passive transfer of colostral immunoglobulins in calves

Passive transfer of colostral immunoglobulins has long been accepted as imperative to optimal calf health. Many factors, including timing of colostrum ingestion, the method and volume of colostrum administration, the immunoglobulin concentration of the colostrum ingested, and the age of the dam have been implicated in affecting the optimization of absorption. The practice of colostrum pooling, the breed and presence of the dam, and the presence of respiratory acidosis in the calf also may affect passive transfer. Various tests have been reported to accurately measure passive transfer status in neonatal calves. The radial immunodiffusion and the enzyme-linked immunosorbent assay (ELISA) are the only tests that directly measure serum IgG concentration. All other available tests including serum total solids by refractometry, sodium sulfite turbidity test, zinc sulfate turbidity test, serum gamma-glutamyl transferase activity, and whole blood glutaraldehyde gelation estimate serum IgG concentration based on concentration of total globulins or other proteins whose passive transfer is statistically associated with that of IgG. This paper presents a comprehensive review of the literature of passive transfer in calves including factors that affect passive transfer status, testing modalities, effects of failure of passive transfer on baseline mortality, consequences of failure of passive transfer, and some treatment options. Many previously accepted truisms regarding passive transfer in calves should be rejected based on the results of recent research.     

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.     

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 refractometers and test endpoints in the measurement of serum protein concentration to assess passive transfer status in calves

OBJECTIVE: To evaluate 3 refractometers for detection of failure of passive transfer (FPT) of immunity in calves, and assess the effect of refractometric test endpoints on sensitivity, specificity, and proportion of calves classified correctly with regard to passive transfer status. DESIGN: Prospective study. ANIMALS: 90 calves. PROCEDURE: Blood samples were obtained from calves that were < 10 days old. Serum IgG concentration was determined by use of a radial immunodiffusion assay. Accuracy of 3 refractometers in the prediction of serum IgG concentration was determined by use of standard epidemiologic methods and a linear regression model. RESULTS: At a serum protein concentration test endpoint of 5.2 g/dL, sensitivity of each refractometer was 0.89 or 0.93, and specificity ranged from 0.80 to 0.91. For all refractometers, serum protein concentration test endpoints of 5.0 or 5.2 g/dL resulted in sensitivity > 0.80, specificity > 0.80, and proportion of calves classified correctly > 0.85. Serum protein concentrations equivalent to 1,000 mg of IgG/dL of serum were 4.9, 4.8, and 5.1 g/dL for the 3 refractometers. CONCLUSIONS AND CLINICAL RELEVANCE: The refractometers, including a nontemperature-compensating instrument, performed similarly in detection of FPT. Serum protein concentration test endpoints of 5.0 and 5.2 g/dL yielded accurate results in the assessment of adequacy of passive transfer; lower or higher test endpoints misclassified larger numbers of calves.     

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.     

Evaluation of assay procedures for prediction of passive transfer status in lambs

OBJECTIVE: To compare 4 assay procedures for prediction of passive transfer status in lambs. ANIMALS: Thirty-one 1-day-old Sardinian lambs. PROCEDURE: Serum IgG concentration was determined by use of single radial immunodiffusion. The following were determined: serum total protein concentration as measured by refractometry (ie, refractometry serum total protein concentration), serum total protein concentration as determined by the biuret method (ie, biuret method serum total protein concentration), serum gamma-globulin concentration as determined by serum protein electrophoresis, and serum gamma-glutamyltransferase (GGT) activity as measured by spectrophotometry. Accuracy of these assays for estimation of serum IgG concentration in 1-day-old lambs was established by use of linear regression analysis. RESULTS: Refractometry serum total protein concentration, biuret method serum total protein concentration, and serum gamma-globulin concentration were closely and linearly correlated with serum IgG concentration. The natural logarithm (ln) of serum GGT activity was closely and linearly correlated with serum IgG concentration (ln). Refractometry serum total protein concentration, biuret method serum total protein concentration, and gamma-globulin concentration accounted for approximately 85%, 91%, and 95% of the variation in serum IgG concentration, respectively. Serum GGT activity (ln) accounted for approximately 92% of the variation in serum IgG concentration (ln). CONCLUSIONS AND CLINICAL RELEVANCE: For prediction of passive transfer status in 1-day-old lambs, serum GGT activity or biuret method serum total protein concentration determination will allow for passive transfer monitoring program development. Immediate refractometry serum total protein concentration determination is beneficial in making timely management and treatment decisions. Serum gamma-globulin concentration determination can be used as a confirmatory test.     

Evaluation of a cow-side immunoassay kit for assessing IgG concentration in colostrum

OBJECTIVE: To determine sensitivity and specificity of a cow-side immunoassay kit for assessing IgG concentration in colostrum. DESIGN: Prospective study. ANIMALS: 76 dairy and 11 beef cows of various parities. PROCEDURE: Colostrum from first, second, and third milkings and milk samples were collected, and IgG concentration was determined by means of radial immunodiffusion. The immunoassay was performed according to the manufacturer's instructions, and sensitivity and specificity were calculated by comparing results of the immunoassay (positive vs negative) with results of immunodiffusion (< 50 g/L vs > or = 50 g/L). RESULTS: 135 colostrum or milk samples were collected. Mean +/- SD colostral IgG concentrations, determined by means of radial immunodiffusion for dairy and beef cows were 65.4 +/- 51.4 g/L and 114.8 +/- 42.7 g/L, respectively. Mean IgG concentrations for first-, second-, and third-milking colostrum samples and for milk samples were 92 +/- 49.0 g/L, 74.6 +/- 45.1 g/L, 47.5 +/- 32 g/L, and 6.8 +/- 3.8 g/L, respectively. Sensitivity of the immunoassay (ie, percentage of samples with IgG concentration < 50 g/L with a positive immunoassay result) was 93%, and specificity (ie, percentage of samples with IgG concentration > or = 50 g/L with a negative immunoassay result) was 76%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that the immunoassay kit was an acceptable cow-side test to identify colostrum samples with IgG concentrations < 50 g/L. The immunoassay kit should be useful in screening colostrum for adequate IgG concentration before feeding to calves or storage.     

Determining the IgG concentrations in bovine colostrum and calf sera with a novel enzymatic assay

Background: Immune protection in newborn calves relies on a combination of the timing,volume and quality of colostrum consumed by the calf after birth.Poor quality colostrum with inadequate immunoglobulin concentration contributes to failed transfer of passive immunity in calves,leading to higher calf morbidity and mortality.Therefore,estimating colostrum quality and ensuring the transfer of passive immunity on farm is of critical importance.Currently,there are no on-farm tools that directly measure immunoglobulin content in colostrum or serum.The aim of this study was to apply a novel molecular assay,split trehalase immunoglobulin G assay(STIGA),to directly estimate immunoglobulin content in dairy and beef colostrum and calf sera,and to examine its potential to be developed as on-farm test.The STIGA is based on a split version of trehalase TreA,an enzyme that converts trehalose into glucose,enabling the use of a common glucometer for signal detection.In a first study,60 dairy and64 beef colostrum and 83 dairy and 84 beef calf sera samples were tested with STIGA,and the resulting glucose production was measured and compared with radial immunodiffusion,the standard method for measuring immunoglobulin concentrations.Results: Pearson correlation coefficients between the methods were determined and the sensitivity,specificity,and accuracy of the test were calculated for different colostrum quality and failed transfer of passive immunity cut-off points.The correlations of the STIGA measured by colorimetric enzymatic reaction compared to radial immunodiffusion for dairy and beef colostrum were 0.72 and 0.73,respectively,whereas the correlations for dairy and beef sera were 0.9 and 0.85,respectively.Next,STIGA was tested in a blinded study with fresh colostrum and serum samples where the correlation coefficient was 0.93 and 0.94,respectively.Furthermore,the performance of STIGA followed by glucometer readings resulted in correlations with radial immunodiffusion of 0.7 and 0.85 for dairy and beef colostrum and 0.94 and 0.83 for dairy and beef calf serum.Conclusions: A split TreA assay was validated for measurement of the immunoglobulin content of colostrum and calf sera using both a lab-based format and in a more user-friendly format compatible with on-farm testing.     

Systematic Review and Meta‐Analysis of Diagnostic Accuracy of Serum Refractometry and Brix Refractometry for the Diagnosis of Inadequate Transfer of Passive Immunity in Calves

Background

Transfer of passive immunity in calves can be assessed by direct measurement of immunoglobulin G (IgG) by methods such as radial immunodiffusion (RID) or turbidimetric immunoassay (TIA). IgG can also be measured indirectly by methods such as serum refractometry (REF) or Brix refractometry (BRIX).

Objectives

To determine the accuracy of REF and BRIX for assessment of inadequate transfer of passive immunity (ITPI) in calves.

Design

Systematic review and meta‐analysis of diagnostic accuracy studies.

Methods

Databases (PubMed and CAB Abstract, Searchable Proceedings of Animal Science) and Google Scholar were searched for relevant studies. Studies were eligible if the accuracy (sensitivity and specificity) of REF or BRIX was determined using direct measurement of IgG by RID or turbidimetry as the reference standard. The study population included calves <14 days old that were fed with natural colostrum (colostrum replacement products were excluded). Quality assessment was performed by the QUADAS‐2 tool. Hierarchical models were used for meta‐analysis.

Results

From 1,291 references identified, 13 studies of 3,788 calves were included. Of these, 11 studies evaluated REF and 5 studies evaluated BRIX. The median (range) prevalence of ITPI (defined as calves with IgG <10 g/L by RID or TIA) was 21% (1.3–56%). Risk of bias and applicability concerns were generally low or unclear. For REF, summary estimates were obtained for 2 different cutoffs: 5.2 g/dL (6 studies) and 5.5 g/dL (5 studies). For the 5.2 g/dL cutoff, the summary sensitivity (95% CI) and specificity (95% CI) were 76.1% (63.8–85.2%) and 89.3% (82.3–93.7%), and 88.2% (80.2–93.3%) and 77.9% (74.5–81.0%) for the 5.5 g/dL cutoff. Due to the low number of studies using the same cutoffs, summary estimates could not be obtained for BRIX.

Conclusions and Clinical Importance

Despite their widespread use on dairy farms, evidence about the optimal strategy for using refractometry, including the optimal cutoff, are sparse (especially for BRIX). When using REF to rule out ITPI in herds, the 5.5 g/dL cutoff may be used whereas for ruling in ITPI, the 5.2 g/dL cutoff may be used.     

Serum IgG Concentrations after Intravenous Serum Transfusion in a Randomized Clinical Trial in Dairy Calves with Inadequate Transfer of Colostral Immunoglobulins

Background: Plasma transfusions have been used clinically in the management of neonates with failure of passive transfer. No studies have evaluated the effect of IV serum transfusions on serum IgG concentrations in dairy calves with inadequate transfer of passive immunity.
Hypothesis: A commercially available serum product will increase serum immunoglobulin concentration in calves with inadequate transfer of colostral immunoglobulins.
Animals: Thirty-two Jersey and Jersey-Holstein cross calves with inadequate colostral transfer of immunoglobulins (serum total protein <5.0 g/L).
Methods: Thirty-two calves were randomly assigned to either control (n = 15) or treated (n = 17) groups. Treated calves received 0.5 L of a pooled serum product IV. Serum IgG concentrations before and after serum transfusion were determined by radial immunodiffusion.
Results: Serum protein concentrations increased from time 0 to 72 hours in both control and transfused calves and the difference was significant between the control and treatment groups ( P < .001). Mean pre- and posttreatment serum IgG concentrations in control and transfused calves did not differ significantly. Median serum IgG concentrations decreased from 0 to 72 hours by 70 mg/dL in control calves and increased over the same time interval in transfused calves by 210 mg/dL. The difference was significant between groups ( P < .001). The percentage of calves that had failure of immunoglobulin transfer 72 hours after serum transfusion was 82.4%.
Conclusions and Clinical Importance: Serum administration at the dosage reported did not provide adequate serum IgG concentrations in neonatal calves with inadequate transfer of colostral immunoglobulins.     

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.     

Factors associated with serum immunoglobulin levels in beef calves from Alberta and Saskatchewan and association between passive transfer and health outcomes

Inadequate consumption of colostrum can negatively affect calf health and survival. The serum immunoglobulin G (IgG) concentrations of 935 beef calves from 152 herds in Alberta and Saskatchewan have been described, using radial immunodiffusion. The determinants and health effects of serum IgG concentrations were studied in 601 calves sampled between 2 and 8 days of age. Of these calves, 6% had failure of passive transfer and an additional 10% had marginal passive transfer. Serum IgG concentrations were lower in calves born to a heifer, as a twin, or experiencing dystocia. The odds of both calf death and treatment were increased in calves with serum IgG concentrations below 24 g/L; a threshold notably higher than the 16 g/L usually considered as providing adequate passive transfer. The finding of 1/3 of calves with serum IgG concentrations less than 24 g/L suggests that calfhood treatments and mortality could be decreased by ensuring that high risk calves consume colostrum.     

Comparison of four screening techniques for the diagnosis of equine neonatal hypogammaglobulinemia

Using radial immunodiffusion as a standard, 4 screening techniques for detection of failure of passive transfer in equine neonates were compared for sensitivity, specificity, positive and negative predictive values, efficiency, and cost. The techniques compared were latex agglutination test, membrane filter ELISA, dipstick ELISA, and glutaraldehyde coagulation (GC) test. Test results of 50 serum samples from foals 24 to 60 hours old revealed consistently highest accuracy in the GC test at IgG concentrations of 400 and 800 mg/dl, and lowest cost per test, using the GC test. Two hundred fifty-three serum samples from foals 24 to 60 hours old were evaluated for comparison of results of GC and radial immunodiffusion tests. Overall efficiency was 92 and 91% at serum IgG concentrations of 400 and 800 mg/dl, respectively. Under most field circumstances, the GC test would be the preferred screening test for detection of failure of passive transfer in equine neonates.     

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.     

Evaluation of a Whole Blood Glutaraldehyde Coagulation Test for the Detection of Failure of Passive Transfer in Calves

The accuracy of a commercially available whole blood glutaraldehyde clot test in the detection of failure of passive transfer (serum immunoglobulin [Ig]G₁ < 1,000 mg/dL) in neonatal calves was evaluated. Serum samples were obtained from 242 calves ranging in age from 1 to 8 days, and comparisons were made with serum lgG₁ concentrations determined by radial immunodiffusion. Both the sensitivity and specificity of the currently marketed whole blood glutaraldehyde clot test are inadequate for routine diagnostic use. Concerns regarding test sensitivity are the most problematic. Sensitivity varied from 0.41 to 0.00, depending on the choice of test endpoint. Specificity varied from 0.85 to 1.00, depending on the choice of test endpoint. Regression analysis demonstrated that the relationship between serum lgG₁ concentration and the glutaraldehyde clot results, although significant (P< .10), was of negligible biological relevance ( r ² = .034). J Vet Intern Med 1996; 10:82–84. Copyright © 1996 by the American College of Veterinary Internal Medicine .     

A review of diagnostic tests for diagnosing failure of transfer of passive immunity in dairy calves in New Zealand

ABSTRACT

The aim of this review is to critically assess the test characteristics and practicality of published data on direct and indirect tests for diagnosing failure of transfer of passive immunity (FPT) in dairy calves in New Zealand, to provide recommendations for veterinary practitioners, and to examine the recommended sample size for assessing herd-level prevalence of FPT and the confidence in the results obtained. The definition of FPT is based on measurement of concentrations of IgG in serum of neonatal calves after colostrum intake. The gold standard method for measurement of concentrations of IgG is radial immunodiffusion. However its cost, requirements for laboratory equipment, and the time taken to obtain results have meant that alternative tests have been developed. The turbidimetric immunoassay and ELISA also directly measure concentrations of IgG. Indirect tests include measurement of concentrations of total proteins (TP) in the laboratory or using a refractometer, γ-glutamyl transferase (GGT) activity, and the zinc sulfate turbidity (ZST) test. Of the indirect tests, measurement of concentrations of TP in the laboratory or using a refractometer combine high specificity and sensitivity with a consistent association with concentrations of IgG in calves between 1–7 days of age. Using a refractometer is less accurate than direct measurement in a laboratory, but is still a suitable test if low cost and speed are important. Although GGT activity is strongly associated with concentrations of IgG in serum, the relationship varies with time after birth. Therefore the target thresholds change with time, increasing error compared to the measurement of concentrations of TP in serum. Similarly, factors other than total concentrations of IgG have a significant effect on the association with ZST test, complicating interpretation. Thus, when direct measurement of concentrations of IgG is not feasible, the recommendation is that concentrations of TP in serum should be used as the diagnostic test for diagnosis of FPT, providing calves are not dehydrated. Using a sample size of 12 calves is suitable for estimating whether the herd-level prevalence of FPT is <20% or >20%, if there are no calves or >5 calves diagnosed with FPT, respectively, but is limited in diagnostic confidence when 1–4 calves test positive. Diagnostic interpretation can be significantly improved if tests of FPT are used alongside information on the likely risk of FPT on the tested farm.     

Effect of colostrum administration by use of oroesophageal intubation on serum IgG concentrations in Holstein bull calves

OBJECTIVE: To determine the amount of colostral IgG required for adequate passive transfer in calves administered colostrum by use of oroesophageal intubation and evaluate the impact of other factors on passive transfer of colostral immunoglobulins in calves. ANIMALS: 120 Holstein bull calves. PROCEDURES: Calves were randomly assigned to specific treatment groups on the basis of volume of colostrum administered and age of calf at administration of colostrum. Colostrum was administered once by oroesophageal intubation. Equal numbers of calves received 1, 2, 3, or 4 L of colostrum, and equal numbers of calves received colostrum at 2, 6, 10, 14, 18, or 22 hours after birth. Serum samples were obtained from calves 48 hours after birth for IgG determination by radial immunodiffusion assay. Effects of factors affecting transfer of colostral immunoglobulins were determined by use of a stepwise multiple regression model and logistic regression models. RESULTS: A minimum of 153 g of colostral IgG was required for optimum colostral transfer of immunoglobulins when calves were fed 3L of colostrum at 2 hours after birth. Substantially larger IgG intakes were required by calves fed colostrum > 2 hours after birth. CONCLUSIONS AND CLINICAL RELEVANCE: Feeding 100 g of colostral IgG by oroesophageal intubation was insufficient for adequate passive transfer of colostral immunoglobulins. At least 150 to 200 g of colostral IgG was required for adequate passive transfer of colostral immunoglobulins. Use of an oroesophageal tube for administration of 3 L of colostrum to calves within 2 hours after birth is recommended.     

Measurement of serum IgG in foals by radial immunodiffusion and automated turbidimetric immunoassay

Hypogammaglobulinemia as a result of failure of transfer of passive immunity (FTPI) is an important risk factor for infectious disease in neonatal foals. The current gold standard for determining serum immunoglobulin concentrations is radial immunodiffusion (RID). The purpose of this study was to compare immunoglobulin concentrations measured by RID with those determined by an automated turbidimetric immunoassay (TIA), which has a much shorter turnaround time. Immunoglobulin concentrations were measured by both RID and TIA in serum collected from 84 neonatal foals. Sixty-seven foals had results within the linear range for both assays. Sensitivity and specificity of TIA for diagnosis of FTPI with IgG < or = 800 mg/dL were 0.81 (95% CI 0.70-0.88) and 0.86 (95% CI 0.76-0.93) and with IgG < or = 400 mg/dL were 0.63 (95% CI 0.35-0.86) and 0.92 (95% CI 0.87-0.95), respectively. A significant linear relationship was found between IgG concentrations determined by TIA and RID (TIA = 0.9511RID + 8.4354; R2 = .59, P < .0001). The coefficients of variation for between-run and within-run precision for the TIA were 2.5 and 3%, respectively. Storage of samples from 10 foals at -20 degrees C for 10-12 months resulted in a reduction in TIA-measured serum IgG concentration of -17.6% (SD = 3.7%), indicating that long-term storage of samples at -20 degrees C should be avoided. The results of this study indicate that measurement of serum IgG by TIA can be used to evaluate foals for FTPI.