ACTIVITY OF CREATINE-PHOSPHOKINASE IN THE COURSE OF EXPERIMENTAL TRICHINELLOSIS

Schanzel H., E. Hegerova, B. Koudela, V. Lohr: Activity of Creatine-Phosphokinase in the Course of Experimental Trichinellosis. Acta vet. Brno, 47, 1978: 91-95. Activity of serum cteatine-phosphokinase has been determined at different intervals after experimental infestation of mice and guinea pigs by larvae of Trichinella spiralis. Blood samples were treated by Bio-La-test kreatinkinaza Lachema, measurements w¢re performed on the spectrophotometer UV VIS at 400 nm wavelength. Intensity of parasitic infestation was estimated by the number of larvae in 1 g of muscle tissue. No significant difference from control values was found in infested animals at any stage of trichinellosis. Determination of CPK activity seems not to be convenient for intravital detection of trichinellosis. Guinea pigs, mice, serum CPK, muscle. Determination of activity of plasmatic creatine-phosphokinase developed to a favourite method for investigation of various myositic processes in man and animals. It not only became a routine method for diagnostics of heart attack Pojer a. o. (1963), Gerber (1965), Ninger (1968), but, it has been studied in connection with muscular dystrophy and other neuromuscular disorders Okineka and Kumagai (1961), Pearce a. o. (1964), Hatnarska a. o. (1968), with degenerative myopathy in young cattle Gils and Zayed (1966), Dotta and Robutti (1972), Martig a. o. (1972), McMurray and McEldowney (1977), with muscular dystrophy in pigs Steinhauser and Rochova (1977), with tetanus Irwin (1967), with hypothermia and stress Meltzer (1971), with muscular affection due to intoxications and infections Oldershausen a. o. (1965), with fatal intoxication by carbon monoxide Bour a. o. (1962), with extraordinary physical exhaustion and hypoxia Cunningham and Critz (1972). Considering the role of creatine-phosphokinase in physiology and pathology of the muscle, we took it for justified to investigate whether changes in CPK activity would occur in consequence of migration and settlement of larvae of Trichinella spiralis. Prior to the trial, physiological values for CPK activity were to be determined in mice and guinea pigs, since the two species were intended to be used for infestation Schanzel and Hegerova (1978). Numerous interactions between host and larva of T. spiralis have been known for long. According to Borchert (1954), the attacked muscle shows decrease in total nitrogen, creatine, purine bases, and increase in water, lactic acid, volatile fatty acids, ammonia and products of muscle decay. An enzymatic response by increased phosphatase activity has been demonstrated by Schanzel and Holman (1966). Material and Methods A total of 160 mice and 19 guinea pigs were infested by viable larvae of Trichinella spiralis, obtained by digestion method from experimentally infested rats. Approximately 200 larvae were administered orally to each mouse and about 1000 larvae to each guinea pig. Successively, 10 in-


Material and Methods
A total of 160 mice and 19 guinea pigs were infested by viable larvae of Trichinella spiralis, obtained by digestion method from experimentally infested rats.Approximately 200 larvae were administered orally to each mouse and about 1000 larvae to each guinea pig.Successively, 10 in-fested mice were used for determination of serum CPK activity at days 2, 3, 4, 5, 6, 8, 10, 11, 12, 14, 21, 28, 35, 41, 49 and 56 p. i. Simultaneously with each infested group, eight control mice were examined.
Blood samples were collected by exsanguination of mice.Samples from very small animals were pooled by 2-3, to obtain the needed amount of serum.The samples were treated with Bio-La-test Lachema in the way described by the producer, but, the amounts of components had to be doubled in order to obtain a voluine sufficient to fill the measuring cell.Determinations of CPK activity were performed on the spectrophotometer UV VIS at 400 nm wavelength, with automatical recording of extinction.
Blood samples from guinea pigs were collected by heart puncture, immediately centrifuged and processed in the same way as samples from mice.The punctures were repeated twice or three times in each guinea pig, first at day 42, and than at day 156 and 197 p.i.
Following determination of CPK activity, the experimental animals were killed.Samples of 1 g muscle tissue -mm.masseteri from guinea pigs and mm.quadricipies and mm.longissimi dorsi from mice -were microscopically examined for the number of larvae of T. spiralis.
Limits of confidence were calculated for statistical evaluation of results.

Results
The activity of serum creatine-phosphokinase in control mice and at different intervals p. i. with T. spiralis is illustrated in Figure 1.
In Table 1, the levels of CPK activity and the number oflarvae/g muscle tissue are compared.
Table 2 demonstrates the same correlation in individual mice, examined 6 weeks p. i.
Proportion between CPK activity in guinea pigs, examined repeatedly at different intervals p. i., and number of larvaejg of their muscle tissue are summarized in Table 3 Not even in this group there was a correlation between CPK activity and intensity of infestation by larvae of T. spiralis.Table 1 shows CPK activity and number of larvae/g muscle tissue group by group, Table 2 the same values separately for each animal in the group examined 42 days p. i.It is obvious from both tables than neither a direct nor an indirect correlation could be established between CPK activity and the number of parasitic larvae.
The results with infested guinea pigs are summarized in Table 3.Here again is to be seen that there is no significant difference between CPK activity in infested and control animals, and, that there is no correlation between CPK activity and intensity of infestation in the course of trichinellosis.
The conclusion resulting from our trial is that the expected effect of larvae of Trichinella spiralis on the activity of creatine-phosphokinase in blood serum of the host could not be observed.Infestation by a larger number of larvae would Day p.i.
a significantly elevated CPK-activity.Determination of CPK activity is thus not a convenient subsidiary method for detecting trichinellosis intravitally.
Nevertheless, the limits of confidence demonstrated in Fig.1make it obvious that no such interpretation can be justified.Only the 10 animals examined 42 days p. i. formed a group differring from values in normal mice just on the border of significance (P = 0.05).The CPK activity in blood serum of infested mice was 160.98 ± 41.29, while the value in control mice was 82.67 ± 31.13 lUlL.

Table 3
Correlation between CPK activity and number of T. spiralls larvae/g muscle tissue in mice 6 weeks p. 1.CPK activity and number of T. spiralis larvae in guinea pigs at different age