ISRAEL JOURNAL OF

VETERINARY MEDICINE
VOLUME 54 (1), 1999

TITLE: SERUM IMMUNOGLOBULIN CONCENTRATIONS IN DIARRHEIC CALVES AND THEIR MEASUREMENT BY SINGLE RADIAL IMMUNODIFFUSION AUTHOR: A. Basoglu, I. amkerten and M. Servinc C Department of Internal Medicine, Faculty of Veterinary Medicine, Selcuk University, 42020 imenlik Konya, Turkey C

Abstract
The effect of passive immune status in diarrheic calves was evaluated. Serum immunoglobulin G (IgG) levels were measured by single radial immuno- diffusion (SRID) test. There was a significant difference in IgG levels (p<0.01) between the healthy and diarrheic groups. It was observed that IgG levels were important in calf diarrhea while diarrhea occurred in calves with partial or normal passive transfer of IgG. SRID test for immunoglobulins was a practical and quantitative field test.

Introduction
Neonatal calf diarrhea is one of the most devastating diseases of the dairy industry, with an estimated incidence as high as 10 to 15% and morbidity approaching 100% in severely affected herds (1). The passive immune status at 24 h. postpartum was an important determinant of health before and after weaning (2). Nevertheless, information gathered from large study populations has demonstrated that healthy, productive neonates with low serum immunoglobulins. Several indirect and direct methods of detecting failure of passive transfer have been described, while the direct measurement of serum IgG concentrations is usually performed using SRID (3). The objectives of this study were to determine IgG levels during the neonatal period, to emphesize the role of IgG in calf diarrhea and to evaluate the SRID test.

Materials and Methods

Twenty healthy and 20 diarrheic day old calves were used in this study. After clinical and hematological examination, serum IgG levels were measured by SRID test and on days 2, 3 and 10 in the healthy calves. A commercially available kit (Bovine SRID Kit-VMRD Products, Pullman, Wash. USA) was used for the IgG determination. SRID plates containing agarose gel incorporating anti-bovine IgG were used. A 3 l micropipette was used to add reference and test samples to the plate wells. The plates were left undisturbed for 18 hours at room temperature after adding the samples. The resulting ring diameters were used to quantitatively estimate the IgG content of the particular serum using regression analysis. A standard curve was established using reference sera supplied by the manufacturer. Test samples and specimens with diameters beyond the range of the standard curve were repeated with additional dilutions. The mean serum immunoglobulin concentrations of the sick and healthy were compared using Students t-test.

Results
Clinical and hematological findings in diarrheic calves were as follows: B.W (Kg) 30 Faecal color+cons Yellowish Watery Age (Days) 9.5 Ht (%) 50.3 B.T. (oC) 38.5 Hb (8.03) 8.03 Heart Rate 131.3 RBC (/l) 7114 Resp./min 50.39 WBC (/l) 9640 Dehyd. ration % 8.1 pH 7.021

Calves were classified as having failure (<800 mg/dl), partial failure (8001600 mg/dl), or normal passive transfer (>1600 mg/dl) on the basis of their serum IgG concentration (2,3). The diarrheic group had partial failure of passive transfer (367.989.7, <800 mg/dl in 35%; 1414.376..9, 800-1600 mg/dl in 35% and 1375.0201.6, >1600 mg/dl in 30%), while the healthy group had normal passive transfer (1220.071.4, 800-1600 mg/dl in 45% and 2786.4227.8, >1600 mg/dl in 55%). Thus, there was a significant difference in IgG levels (p<0.01) between healthy and diarrheic groups. During the neonatal period there were no significant differences in IgG levels between calves at two to three days of age and at ten days of age.

Discussion
Despite improved husbandry and hygiene, infectious disease remains the leading cause of morbidity and mortality in neonatal calves over three days old. Numerous publications in the past three decades correlated neonatal morbidity and mortality rates with low levels of serum immunoglobulins or what has been popularly termed failure of passive transfer in calves (2, 3-5). At this point, there are many contrasting opinions: Serum immunoglobulin concentration is only one component of the neonatal complex immune system, while hypogammaglobulinemia does not always result in infection or death. Similarly, overwhelming sepsis can develop in neonates that ostensibly received adequate passive transfer; thus normal serum immunoglobulin levels are not synonymous with immunity (6). In the present study, it was observed that IgG levels were important in occurrence of calf diarrhea, while the disease was also observed in calves with partial and normal passive transfer. In sera, colostral immunoglobulin levels peak at 24-48 hours thereafter declining, each class according to its half-life until approximately 3 weeks of age when the IgG levels begin to rise as the calf synthesizes immunoglobulin (7). In calves that acquire adequate passive immunity after birth, the globulin concentration drops to its nadir at about 2 months of age (8). Calves can be tested for immune status when they are between 24 hours and 7 days of age. In this study, while healthy 2 to 3 days old calves had a partial or normal passive IgG transfer, they had partial and inadequate passive transfer by the 10th day. The determination of serum immunoglobulin concentrations is the most accurate means of assessing the passive immune status; however, the cost, technical expertise, and the relevance of the results after 24 to 48 hours required for running the test is a limiting factor. Several indirect methods of determination are also available. These include zinc sulfate turbidity, sodium sulfate precipitation, glutaraldehyde test, and serum refractometry (3). The traditionally reliable SRID test, when accurately performed, can be used as to screen which calves have absorbed sufficient immunoglobulin (4). In this study also, the ability to identify failure of passive transfer in calves by evaluating serum IgG concentration was good, and the SRID test was an easy, practical and quantitative means of measurement. In conclusion, however, IgG levels were important in occurrence of calf diarrhea, the disease was also observed in calves with partial or normal passive transfer, and SRID test for IgG measurements was practical and a quantitative field test.

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