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

Abstract

The accuracy of a commercially available whole blood glutaraldehyde clot test in the detection of failure of passive transfer (serum immunoglobulin [IglG, <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 lgG, 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 alves are born with negligible concentrations of immu-C noglobulin (Ig). Ig requirements are met by ingesting and absorbing the colostral 1gs.l.' This process, termed passive transfer, is not the result of an active immune response by the neonate. In this regard, passive transfer of Ig is a critical determinant of calf health.' Although colostra vary with regard to their Ig concentration, the timing and volume of colostrum intake also are critical in assuring adequate passive transfer of Ig; colostral Ig absorption is best at birth and declines within hours. ' Routine monitoring of passive transfer in calves can be used to assess management and husbandry practices. Test results can be used to critique calf rearing practices and develop intervention strategies to optimize calf health and farm productivity. In calves of exceptional value or genetic merit, quantification of serum Ig concentration permits identification of calves which require Ig supplementation. Tests of this type also are useful in a clinical setting because they assist with the diagnosis and prognosis of individual sick neonates.A variety of assay procedures have been developed to measure serum Ig concentrations in neonatal livestock. These include refractometer determination of total serum protein concentration, zinc sulfate and sodium sulfite turbidity assays, immunoassays, and the glutaraldehyde gelation assay.',' Any assay procedure used should be readily adapted to on-farm application. Immediacy of results permits feedback to calf rearing personnel, and creates incentives for optimal management practices. Additionally, rapidly available test results may permit targeted treatment of individual calves. Consequently, test procedures should provide rapid and reliable results, require minimal instrumentation, and be inexpensive.A whole blood glutaraldehyde gelation assay which apparently addresses many of these concerns has recently been marketed. In this assay, whole blood is added to a reagent tube containing glutaraldehyde and the time required for clot formation is measured. Gelation times less than 5 minutes are considered indicative of adequate passive transfer. Because the test is performed on whole blood rather than serum, this test has advantages relative to turbidity tests and refractometry, which require instrumentation (eg, centrifuge, temperature-compensated refractometer), and sample processing before a test can be performed.Use of glutaraldehyde gelation of Ig as an assay for serum Ig concentration is hardly a no...

Bovine Neonatal Immunology

“…Recently, several articles have been published that have fine-tuned these various testing methods to provide more accurate values for defining immunoglobulin levels in the neonate. 25,47,65,66 Although the identification of optimum threshold values for these tests is useful, they must be kept in perspective and their use not overinterpreted, Various studies that have examined the relationship between serum immunoglobulin levels in calves and disease incidence and subsequent production have had conflicting results. Several studies have demonstrated significant increases in morbidity and mortality in calves suffering failure of passive transfer and have been reviewed.…”

Preliminary validation of a calf-side test for diagnosis of failure of transfer of passive immunity in dairy calves

“…Imprecision in RID assay replicates from the same sample have been noted and are attributed preliminary validation of a calf-side test for diagnosis of failure of transfer of passive immunity in dairy calves I. elsohaby * † 1 and G. p. Keefe * 4755 to inconsistencies in the assay standards (Ameri and Wilkerson, 2008). Other than RID, serum total protein determination using refractometry, sodium sulfite turbidity test, zinc sulfate turbidity test, ELISA, serum γ-glutamyl transferase activity, and whole-blood glutaraldehyde coagulation test have been described with varying degrees of accuracy for predicting IgG concentrations in calves (Tyler et al, 1996a;Parish et al, 1997;Weaver et al, 2000). The turbidimetric immunoassay described by Etzel et al (1997) performed well in calves (Dawes et al, 2002;Alley et al, 2012) and foals (Davis et al, 2005).…”

“…Several assays are available to assess passive immunoglobulin transfer status in newborn calves (Tyler et al, 1996a;Parish et al, 1997;Weaver et al, 2000). The radial immunodiffusion (RID) assay is the reference standard method for determining passive immunoglobulin transfer by measuring the quantity of IgG in the serum (Weaver et al, 2000;Godden, 2008).…”

Differences in Total Protein Concentration between Fresh and Frozen Serum and Plasma Samples Used to Assess Failure of Passive Transfer in Dairy Calves

“…Several methods have been reported to evaluate passive transfer status in neonatal calves including zinc sulfate and sodium sulfite turbidity tests [10,11], glutaraldehyde coagulation tests [12], measurement of serum gamma-glutamyl transferase (GGT) activity [13,14], and direct measurement of IgG through single radial immune diffusion (SRID), which is considered the gold standard [11,13,15]. Serum total protein (TP serum ) concentration has been shown to correlate with serum IgG concentration [10,16].…”