Localization and characteristics of lactate and malate dehydrogenase in the sparganum and adult worm of <i>Spirometra erinacei</i>

, Kwak, K H; , Cheon, E W; , Kim, C H

doi:1996.34.1.59

Korean J Parasito > Volume 34(1):1996 > Article

Original Article


Korean J Parasitol. 1996 Mar;34(1):59-68. English. Published online Mar 20, 1996. http://dx.doi.org/10.3347/kjp.1996.34.1.59
Copyright © 1996 by The Korean Society for Parasitology


Localization and characteristics of lactate and malate dehydrogenase in the sparganum and adult worm of Spirometra erinacei
Kee-Hoon Kwak,Eun-Woo Cheon and Chang-Hwan Kim^*
Department of Biology, College of Natural Sciences Gyeongsang National University, Chinju 660-701, Korea.

Received November 27, 1995; Accepted December 22, 1996.
Abstract
This study investigated the enzyme histochemical localization and characteristics of lactate (LDH) and malate dehydrogenase (MDH) related with the oxidation-reduction metabolism in the sparganum and adult of S. erinacei. By enzyme histochemical assay, activity of LDH was strong in the tegument and subtegumental muscle layers of the adult and sparganum. Activity of MDH was strong in the tegument of the sparganum and subtegumental muscle layers of the adult. However it was weak in the tegument of the adult. By electrophoresis, 45 kDa band was major and common in LDH of adults and spargana. The 150 kDa molecule was the major and common band in MDH of adults and r-spargana (from experimentally infected rats). By isoelectrofocusing, isoelectric points (PI) of 4 MDH isozyme from adult worm were 6.0, 6.5, 6.7 and 7.1, respectively. PI 6.0 was the major band. The active range of pH for MDH was about pH 6 approximately 8 and the optimum pH was pH 7. The effective temperature on the MDH was about 30℃ approximately 50℃ and the optimum temperature was about 40℃ in spargana and adult worm. In the stability against heat, when MDH was heated at 85℃ for 10 seconds, the activity was denatured perfectly. Maximum activity of MDH was 19.4 unit in the s-sparganum (from snakes), 24.5 unit in the r-sparganum (from rats) and 108.0 unit in the adult worm. The maximum activity was higher in adults than in spargana. The present result showed us that the nutrients absorbed through the tegument were transferred into inner tissues and were utilized as the source of metabolism. According to the habitat of the parasite, the isozymes of LDH and MDH are activated differently, and by this different activation the sparganum and adult can adapt themselves to parasitic circumstances.

Figures

Fig. 1
Histochemical localization of lactate dehydrogenase on the sparganum and adult worm of S. erinacei. a. Snake sparganum (80 ×), b. Snake sparganum (400 ×), c. Rat sparganum (80 ×), d. Rat sparganum (400 ×), e. Immature proglottid (80 ×), f. Immature proglottid (400 ×), g. Mature proglottid (80 ×), h. Mature proglottid (400 ×), i. Gravid proglottid (80 ×), j. Gravid proglottid (400 ×). T, tegument; SM, subtegumental musculature; PM, parenchymal musculature; CC, calcareous corpuscles; TS, testis; V, vitelline gland; EG, egg; N, nerve trunk; LM, epidermal longitudinal musculature; GP, genital pore.

Fig. 2
Histochemical localization of malate dehydrogenase on the sparganum and adult worm of S. erinacei. a. Snake sparganum (80 ×), b. Snake sparganum (400 ×), c. Rat sparganum (80 ×), d. Rat sparganum (400 ×), e. Immature proglottid (80 ×), f. Immature proglottid (400 ×), g. Mature proglottid (80 ×), h. Mature proglottid (400 ×), i. Gravid proglottid (80 ×), j. Gravid proglottid (400 ×). T, tegument; SM, subtegumental musculature; PM, parenchymal musculature; CC, calcareous corpuscles; TS, testis; N, nerve trunk.


	Fig. 3 Isozyme patterns of lactate dehydrogenase on the sparganum and adult worm of S. erinacei. SS, snake sparganum; RS, rat sparganum; AD, adult; TD, tracking dye; kDa, kilo-dalton; M, marker protein.


	Fig. 4 Isozyme patterns of malate dehydrogenase on the sparganum and adult worm of S. erinacei. SS, snake sparganum; RS, rat sparganum; AD, adult; TD, tracking dye; kDa, kilo-dalton; M, marker protein.


	Fig. 5 Isozyme patterns of malate dehydrogenase by IEF no the sparganum and adult worm of S. erinacei. TP, Coomasie blue staining of total protein(adult); PI, isoelectric point; M, marker protein; AD, adult.


	Fig. 6 The stability against heat treatment on the isozmy of malate dehydrogenase in adult worm, heating at 85℃ for 10, 20, 30, 40s seconds, respectively. 0, control; 10, 10 sec.; 20, 20 sec.; 30, 30 sec.; 40, 40 sec.


	Fig. 7 The effect of pH on the activity of malate dehydrogenase in r-sparganum and adult worm. ●—●, adult; ○—○, sparganum


	Fig. 8 The effect of temperature on the activity of malate dehydrogenase in r-sparganum and adult worm. ●—●, adult; ○—○, sparganum

Tables


	Table 1 Relative amount of lactate dehydrogenase activity on the sparganum adult worm of S. erinacei


	Table 2 Relative amount of malate dehydrogenase activity on the sparganum adult worm of S. erinacei


	Table 3 Enzyme activities of malate dehydrogenase on the sparganum and adult worm of S. erinacei (mean ± SE)

References


1.	Barrett J. The anaerobic end-products of helminths. Parasitology 1984;88(Pt 1):179–198.

2.	Barry DH, Mawdesley-Thomas LE. Enzyme histochemistry of the adult liver fluke, Fasciola hepatica. Exp Parasitol 1968;23(3):355–360.

3.	Bennet EM, Heath PA, Bryant C. The effects of changes in the definitive host environment on the metabolism of Hymenolepis diminuta during growth and maturation. Int J Parasitol 1993;23(1):57–68.

4.	Bryant C. The regulation of respiratory metabolism in parasitic helminths. Adv Parasitol 1978;16:311–331.

5.	Fukase T, et al. Jpn J Parasitol 1984;33:283–290.

6.	Fukase T, et al. Jpn J Parasitol 1985;34:351–395.

7.	Hotta H, et al. Dept of Medical Zoology Niigata Univ of Med 1978;4:357–368.

9.	Kwak GH, et al. Korean J Zool 1992;35:362–371.

*10.*	Logan J, et al. Comp Biochem Physiol 1977;75B:51–53.

*12.*	Moon TW, et al. Comp Biochem Physiol 1977;56B:249–254.

*13.*	Pappas PW. Membrane transport in helminth parasites: a review. Exp Parasitol 1975;37(3):469–530.

*14.*	Park YC, et al. Korean J Zool 1980;23:263–272.

*15.*	Pappas PW, Schroeder LL. Hymenolepis microstoma: lactate and malate dehydrogenases of the adult worm. Exp Parasitol 1979;47(2):134–139.

*16.*	Rhaman R, et al. Int Biochem J 1973;4:153–162.

*18.*	Walkey M, Fairbairn D. L(+)-lactate dehydrogenases from Hymenolepis diminuta (Cestoda). J Exp Zool 1973;183(3):365–374.

*19.*	Watts SD, Fairbairn D. Anaerobic excretion of fermentation acids by Hymenolepis diminuta during development in the definitive host. J Parasitol 1974;60(4):621–625.