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Immuno-stimulant effects of Ascorbic Acid in Colostrum-deprived neonatal camels

S. I. AL-Sultan1

1 Department of Public Health and Animal Husbandry, College of Veterinary Medicine and Animal Resources, King Faisal University, P. O. Box 11647, Al-Ahsa 31982, Saudi Arabia

ABSTRACT

Deprivation of colostrum has resulted in significantly increased counts of leukocytes, neutrophils and concentration of cortisol and decreased eosinophils, serum ascorbate and lysosomal enzyme activity. Subcutaneous administration of ascorbic acid at a dose of 10mg/kg body weight daily to colostrum-deprived calves has resulted in increased serum ascorbate and lysosomal activity but has no effects on other parameters. It is suggested that colostrum deprivation may act as stressor factors in calves and that ascorbic acid could produce an immuno-stimulant effect in neonatal camel calves.

Keywords: Ascorbic acid, colostrum, immuno-stimulant, camel calves.

INTRODUCTION

The survival of the animal depends on the successful body defense against microbial invasions. The camel calf comes to life almost deprived of serum immunoglobulin and depends on colostrum for virtually all its humoral passive immunity (Ghazi et al., 1994). Failure to achieve adequate transfer of passive immunoglobulin has been associated with increased risk of diarrhea, respiratory affections and motility of neonates (McGuire et al., 1975; Rea et al., 1991).

One of most important vitamins found in colostrum is ascorbic acid. Ascorbic acid is known to enhance immunity in neonates (Dallegri et al., 1980; Thomas and Holt, 1978). This study was carried out to investigate the effects of administration of ascorbic acid on some immune parameters of colostrum–deprived camel calves.

MATERIALS AND METHODS

Animals and treatments

Camel neonates were removed from their dams immediately after birth to prevent suckling and ingesting of colostrum. Neonates were assigned randomly to one of three treatment groups. Group 1 neonates (N=9) were left to suckle their dams. Group 2 neonates (N=9) were colostrums deprived but supplemented with 500 ml camel milk given orally at 2 hours interval from 2 to 20 hours after parturition (total of 5 liters/day). In addition neonates were injected subcutaneously with ascorbic acid at a dose of 10 mg/kg body weight. Group 3 neonates (N=9) were colostrum deprived but given camel milk as in group 2. The treatment continued for 10 days.

Sample collection

Blood samples were collected at day 10 of age into heparinized tubes for hematology or plain tubes for serum which was separated and stored at -20oC until analysis. White blood cell count was determined by a Counter Model 2B1 Counter (Counter Electrics, Hialeah, FL, USA). The differential leukocyte counts were carried out using blood smears stained with Giemsa and May-Grunwald solutions. Serum lysosome concentrations were measured using Micrococcus lysodiekticus substrate (lysosome reagent kit, Warthington Biochemicals, Co.Freehold , NN, USA) according to manufacturers recommendations (AL-Ankari and Homeida, 1996).

The percentage change in transmission at 510 nm per minute was recorded in spectrophotometer (Hatachi, Japan). Serum insulin was estimated with solid phase radioimmunoassay (Ekpe and Christopherson, 2000) using specific kit (Tkco5, Coat-a-Count, Diagnostic Products Corporation, Lost Angeles, CA, USA.). The intra-assay coefficient of variation was 5.4±0.1% and sensitively was 0.2 μg/dL. Serum ascorbate was determined by the method of Okamura (1980). Standard curves for ascorbate was linear over the range of 0 to 10mg/dL. Recovery of added ascorbic acid was 98%. Data obtained was statistically analysed by student t-test.

RESULTS

Results of the effects of colostrums deprivation and treatment with ascorbic acid are summarized in Table 1. Significantly (P<0.05) increased counts of leukocyte, neutrophils and decreased eosinophils were demonstrated in colostrums-deprived calves (group 3) and colostrums-deprived calves treated with ascorbic acid (group 2) compared to controls (group 1).

Table 1. Effects of colostrums deprivation and administration of ascorbic acid on leukocytic counts serum ascorbate and lysosomal enzymes in camel calves at 10 days of age.

Variable	Group 1(N=9) colostrum fed calves (control)	Group 2(N=9) colostrums deprived –calves and treated with ascorbic acids	Group 3(N=9) Colostrums-deprived calves
Leukocytes x109/L	13,200±1,100	19,400±1600*	19,200±1500*
Neutrophils x109/L	4,150±600	10,200±1200*	10,400±1300*
Lymphocytes x109/L	8220±800	7,900±650	8100±600
Eosinophils x 109/L	320±20	22±4*	30±4*
Lysosome activity μ/L	6.3±0.11	6.1±0.12	3.4±0.11*
Ascorbate(mg/dL)	0.45±0.03	1.10±0.11*	0.23±0.04*
Cortisol μg/dL	0.65±0.03	1.93±0.09*	2.13±0.11*

P<0.05 significantly different from controls.

DISCUSSION

Deprivation of colostrum has resulted in increased leukocytes count, neutrophilia and eosinopenia with little effect on lymphocyte counts suggesting that leukocytosis was mainly due to neutrophilia. Deprivation has resulted in an increased in cortisol and decreased in ascorbate and lysosomal activity. Similar observations on levels of ascorbate and cortisol in neonatal calves have been reported (Salageanu et al., 1971; Wegger and Moustgaard, 1982; Cummins and Brunner, 1991) as results of parturition or disease being stressors. The effect on leukocyte count, ascorbate concentration and decreased lysosomal activity may be ascribed to increased cortisol activity as stressor factor as a result of colostrums deprivation. A link between adrenal glucocorticoid secretion and ascorbate release and uptake has been established (Kipp and Rivers, 1987). Glucocorticoids or stress have been reported to cause leukocytosis, neutrophilia, esosinopenia and decreased ascorbic content of neutrophils (Crandon et al., 1961; Roth and Kaeberle, 1985; Cummins and Brunner, 1991). Serum lysosome activity was considered to be an index of macrophage function (Currie and Eccles, 1976). Similar studies showed that methyl palmitate was associated with reduction of lysosome enzyme release in serum (Koskoshis and Diluzio, 1979). Furthermore, dexamethasone has been shown to impair leukocytic phagocytic function (Jain, 1986; Ross et al.,, 2003).

Administration of ascorbic acid has no effect on leukocyte counts or cortisol levels in colostrums-deprived calves. Similar effects have been reported in dexamethazone –treated cattle (Roth and Kaeborle, 1985). However the injection of ascorbic acid has effectively increased lysosomal concentration, suggesting that the vitamin may produce an immuno-stimulant effect in camel neonates. Indeed, dietary ascorbate supplementation did increase plasma immunoglobulin and stimulate cellular and humoral immunity in a variety of species, indicating a direct effect of ascorbate on the function of cells of the immune system (Li and Lovell, 1985; Prinz et al., 1980; Cummins and Brunner, 1991).

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First Published February 2008

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