Some studies on Stiff Lamb Disease in Qassim region in Saudi Arabia.
1: Enzymatic pro®le in free, subclinically and clinically affected
lambs both before and after treatment with
vitamin E and selenium preparation
T.K. El-Neweehy
*, A.A. Al-Qarawi, H.A. Abdel-Rahman
Department of Veterinary Medicine, King Saud University, PO Box, 1482 Bureydah, Saudi Arabia
Accepted 14 July 1999
Abstract
On studying the enzymatic pro®le of serum muscle speci®c enzymes (ALT, AST, LDH and CPK) in two sheep ¯ocks one free while the other was endemic with stiff lamb disease it was found that: (1) Serum muscle speci®c enzymes levels were signi®cantly increased in lambs of the endemic ¯ock when compared with those from the non-endemic one. (2) Vitamin E and selenium administration was followed by signi®cant decrease in enzymes levels in subclinically affected lambs of the endemic ¯ock, while no changes occurred in lambs of the control, non-endemic ¯ock one-week after administration. (3) Lambs in the endemic ¯ock were born with relatively high serum enzyme levels, which began to increase signi®cantly and gradually from the second week of life and up, reaching maximum levels at the fourth week of age. At this age, most of the lambs began to show typical symptoms of the disease. (4) Clinical improvement accompanied with pronounced drop in levels of these enzymes occurred after treatment with vitamin E and selenium preparation in clinically affected lambs.# 2000 Elsevier Science B.V. All rights reserved.
Keywords:Stiff lamb disease; Vitamin E±selenium preparation; Muscle speci®c enzymes levels
1. Introduction
It is now evident that several diseases of farm animals are caused by or associated with de®ciency of selenium and/or vitamin E. Nutritional muscular dystrophy is one of these diseases which is of eco-nomic importance particularly in domestic ruminants (Gallo-Torres, 1972; Hadlow, 1974). Subacute enzoo-tic muscular dystrophy (stiff lamb disease, SLD) is the
most common form of this disease in young lambs (Radostitis et al., 1995).
Characteristic signs of clinical cases of SLD sug-gest the underlying problems of vitamin E and/or selenium de®ciency. Subclinical de®ciencies are more dif®cult to recognize. Early metabolic disturbances can be diagnosed by biochemical means. Activity of muscle speci®c enzymes in the serum show marked increase above normal before clinical disturbances occur (Tollersrud, 1971; Farrell and Olson, 1973; Feldheim, 1973; Radostitis et al., 1995).
*Corresponding author.
Only the administration of vitamin E and selenium is considered to be a suitable treatment for nutritional muscular dystrophy (NMD). Sometimes NMD responds to vitamin E alone and in other cases to selenium. Under ®eld conditions it is clearly impos-sible to determine whether, in a given case, the chance of success is better with vitamin E or selenium; for this reason it must be assumed that a combination of vitamin E and selenium offers the best chance of a positive effect (Adams, 1972).
Recent experiments showed that serum enzymes diagnosis is capable of detecting subclinical stage of this de®ciency (Bostedt, 1976; EL-Neweehy, 1982; Radostitis et al., 1995).This disease may represent part of problem of locomotion disturbance among lambs in our clinic. This study planned to throw some light on the enzymatic pro®le in SLD-free, subclinically and clinically affected Najdi lambs before and after treat-ment with vitamin E and selenium preparation in Qassim region in Saudi Arabia.
2. Materials and methods
A total number of 58 suckling native Najdi lambs belonging to two sheep ¯ocks were used in the present investigation. They were classi®ed into three groups. The ®rst group was of 20 apparently healthy suckling lambs. These lambs was chosen from a ¯ock with no history of frequent occurrence of SLD to be used as control group.
The second group, was of 18 apparently normal lambs but was chosen from other ¯ock endemic with SLD and was suspected to be subclinically affected. Another 20 lambs forming the third group were chosen from the same endemic ¯ock. Lambs of the third group were put under both clinical and bio-chemical observation at weekly intervals from birth till weaning. At about the fourth week of age, most lambs of this group began to show locomotion dis-turbances in the form of stiffness, dif®culty standing and ataxic movement, with pronounced muscle tre-mors especially when forced to stand for long periods. Some showed dyspnea, with abdominal type of respiration. All were of good appetite when helped to reach the dams. These lambs were suspected to be affected with SLD which was con®rmed later enzymatically.
Lambs of both the ®rst and second groups, were given a single dose (3 ml) of vitamin E and selenium preparation (Injacome E±Selenium, Roche, Switzer-land; each millilitre contains 150 mg DLa-tocopherol acetate0.5 mg sodium selenite pentahydrate) at 21 days of age. Four daily successive similar doses were given to clinically affected lambs of the third group. Blood samples of the ®rst and second groups were collected twice only, one at 21 days of age before treatment and the second one week after treatment. Lambs of the third group were sampled at weekly intervals from the ®rst week of life until the appear-ance of signs of the disease (at the 4th week). Another sample was collected one week after the last dose of the treatment trials (at the 5th week). Blood samples were collected by jugular venipuncture. The blood was immediately centrifuged at 3000 g for 10 min to harvest the serum. All the above mentioned serum samples were used for determination of serum muscle speci®c enzymes.
Serum levels of alanine transaminase (ALT), aspar-atate transaminase (AST), lactic dehydrogenase (LDH) and creatine phosphokinase (CPK) were mea-sured by standard methods (Reitman and Frankel, 1957; Wroblewski and Duean, 1955; Forester, 1970, respectively) using commercial diagnostic kits (bio-Merieux, France). The samples showing absorbance for ALT > 125, AST > 165, LDH > 0.01 and CPK > 0.25 were diluted with normal saline and reassayed.
Statistical analysis of the present data was per-formed using the Student-ttest according to Snedecor and Cochran (1976).
3. Results
Table 1 shows that the level of serum muscle speci®c enzymes were signi®cantly high (p< 0.0001) in Najdi lambs from endemic ¯ock when compared with those from non-endemic ones.
Table 2 illustrates that there is gradual increase (p< 0.001) in serum enzyme levels from the second week of life and reaches maximum level (p< 0.0001) at the fourth week of age. Vitamin E and selenium administration was followed with marked decrease in enzyme level one week later at 5th week of age (to levels of one week of age). Similar effects
Table 1
Serum muscle speci®c enzymes (iu/L) in normal (Group 1) and in SLD subclinically affected (Group 2) Najdi lambs before (pre) and one week after treatment (wk) with vitamin E± selenium preparation (values are meanSE)
No. ALT AST LDH CPK
Pre wk Pre wk Pre wk Pre wk
Control (Group 1) 20 7.441.78 7.881.26 25.842.66 20.752.84 678.0261.42 618.24284.5 37.3813.8 33.838.46 SLD subclinical
(Group 2)
18 52.118.6** 22.06.6** 468.46168.2** 68.814.6** 1814.18354.0** 686.8253.53* 1186.45364.6** 122.8414.36**
*P< 0.05 vs. control. **P< 0.0001 vs control.
El-Ne
weehy
et
al.
/
Small
Ruminant
Resear
ch
35
(2000)
219±223
were recorded in SLD subclinically affected lambs (Table 1).
4. Discussion
Serum muscle speci®c enzymes were high in Najdi lambs from an endemic SLD ¯ock when compared with those from a non-endemic one, a ®nding which may indicate they were subclinically affected with stiff lamb disease. (Bostedt, 1976; EL-Neweehy, 1982; Radostitis et al., 1995). Similar results were obtained by Kiroloss et al. (1985) in subclinically affected NMD lambs in Egypt. On the other hand, while vitamin E and selenium administration led to signi®cant decrease in enzymes level in subclinically affected lambs, their levels were unchanged in treated lambs of the control NMD free ¯ock. These ®ndings agree with those reported by Forenbacher et al. (1975) and Radostitis et al. (1995).
The behavior of muscle enzymes levels in lambs of SLD-endemic ¯ock from birth and upwards (Table 2) showed gradual increase in their levels from second week of life and reached their maximum levels at the 4th week of age. At this age most lambs began to show typical signs of SLD. These ®ndings were nearly similar to those reported by Tollersrud (1971) who noted that the activity of muscle speci®c enzymes in the serum in lambs showed marked increases above normal before clinical disturbance occurred. Admin-istration of vitamin E and selenium preparations was accompanied with both improvement of clinical signs of the disease and marked decrease in serum muscle speci®c enzymes level, one week after treatment. These ®ndings were nearly similar to those reported
by Adams (1972), Forenbacher et al. (1975), Bostedt (1976) and Radostitis et al. (1995). Bostedt (1976) reported signi®cant elevation in muscle speci®c enzymes (ALT, AST, LDH, CPK) in lambs of an endemic ¯ock although they were clinically healthy and a return to normal levels within one week after single injections of vitamin E and selenium.
From the above mentioned study, it could be con-cluded that the subclinical form of SLD, could be detected by screening of serum muscle enzymes. In our study, SLD occurred naturally among Najdi lambs around the 4th week of age. Vitamin E and selenium administration was not only accompanied with improvement of clinical signs of the disease but also with signi®cant decrease in serum muscle speci®c enzymes level in both clinical and subclinical forms of the disease.
References
Adams, C.R., 1972. Vitamin E for beef cattle. Cattle Science Handbook 9, 80±85.
Bostedt, H., 1976. Serum enzymaticshe untersuchungen bei jammern im Alter 10±30 tagegleicheitig in Beitrag zur prophylaxe der enzootischen Muskeldystrophie. Berl Munch. Tieraztl. Wschrt Publication.
El-Neweehy, T.K., 1982. Studies on white muscle disease in suckling Egyptian buffalo calves in ARE. Ph.D. Thesis, Vet. Med. Dep., Fac. Vet. Med., Cairo.
Farrell, P.M., Olson, R.E., 1973. Creatine phosphate and adenine nucleotides in muscle from animals with muscular dystrophy. Am. J. Physiol. 225, 1102±1106.
Feldheim, W., 1973. Enzyme activity changes in tocopherol de®ciency states. In: Vitamins, vol. 3. Roche, Basel, pp. 51±69. Forenbacher, S., Herceg, M., Feldhofer, S., 1975. Systemic myophathy of fattening cattle caused by lack of Vitamin E. Veterinaski Arhiv. 45, 159±175.
Table 2
Serum muscle speci®c enzymes in Najdi lambs (Group 3) of SLD endemic ¯ock before, at incidence and one week after treatment (n20) (values are meanSE)
Age before incidence Age of incidence (4th week)
One week after treatment (5th week) One week Two weeks Three weeks
ALT (iu/L) 20.98.24 36.64.4** 84.414.6*** 123.534.5*** 34.013.2*
Forester, G. et al., 1970. Creatin-kinase. Bestimmung mit cteatinphosphat als. Substrat. In: Bergmayer, H.U. (Ed.), Methoden der Enzymatischen Analyse, 2 vols., 2ed. German ed. Verlag Chemic Weinhein, p. 755 ff.
Gallo-Torres, H., 1972. Vitamin E in animal nutrition. Int. J. Nutr. Res. 42, 312±323.
Hadlow, J.W., 1974. Myopathies of animals ± a brief view. In: Homburger, F., Sucas, I., Karger, S. (Eds.), Comparative Pathology of the Heart, Advanced Cardiology, vol. 13, Basel, pp. 68±80.
Kiroloss, F.N., El-Kabany, A.W., Abdel Raouf, M., 1985. Early diagnosis and treatment of subclinical nutritional muscular dystrophy (NMD) in lambs. Alex. J. of Vet. Sci. 1, 123± 132.
Radostitis, O.M., Blood, D.C., Henderson, J.A., Aundel, J.H., Gay, C.C., 1995. Veterinary Medicine: A Textbook of the Disease of Cattle,Sheep, Pigs, Goats and Horses, 8th ed. Bailliere Tindall, London.
Reitman, S., Frankel, S., 1957. A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am. J. Clin. Path. 28, 56±58. Snedecor, G.M., Cochran, W., 1976. Statistical methods, 6th ed.
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Tollersrud, S., 1971. Serum enzyme changes in lambs with experimentally induced acute muscular dystrophy. Acta Vet. Scand. 12, 365±374.
