1.4-Bis-trichlormethyl-benzol이 肝디스토마의 形態的 變化에 미치는 影響
Study on the morphological changes of clonorchis sinensis treated by 1,4 Bis-trichlormethyl-benzol
There have been several reports concerning the morphological changes in Clonorchis sinensis due to drug administration. Baba(1944) reported that the mitochondria in various organs of the parasite showed degenerative changes after the administration of antimonial compounds. Takagi(1962) and Sch et al.(1965) also
examined th morphological changes of Clonorchis sinensis using dithiazanine iodide, but they did not examine the mitochondrial changes.
Recently Yokogawa et al.(196a) and Cho et al.(1966) reported that 1,4 Bis0trichlormethy1-benzol was effective in eliminating Clonorchi sinensis in he bile ducts and caused degenerative changes in the worms, primarily in the sexual organs. However, there were no reports concerning the mechanism of the lethal
effect. The present study was designed to discover the morphological changes in the adult worm and in the cellular mitochondria by light mycroscopy as well as by electron microscopy.
Materials and Methods
Metacercaria of Clonorchis sinensis were collected from Pseudorasbora parva brought from the Kim Hae area known as one of the endemic regions of clonorchiasis in Korea.
Rabbits were orally infested with 500 to 1,000 metacercaria of C. sinensis.
1,4-Bis-trichlormethy1-benzol (Hetol) was supplied by Hoechst Co., Frankfurt/Main, Germany.
For the purpose of the experimental test the Hetol was orally administered once during 60 to 80 days after the infestation, and the dosage of administration to seven groups of rabbits were 25, 50, 100, 200, 300, 400, and 500mg/kg of body weight respectively.
C.Sinensis from the bile ducts were collected 24 to 72 hours after the administration of Hetol. All the worms were put into Ringer's Solution and microscopically examined.
For further study of morphological changes with whole mount preparations, worms were fixed in 10 per cent formalin or Schaudin's solution for 12 hours and stained with aceto-carmine, and observed under the light microscope and also 5 micron thick
paraffin sections, were mounted on slides and stained with hematoxylin and eosin, or Heidenhain's-iron hematoxylin.
For the study of electron microscopy, each group was dissected 72 hours after the Hetol administration.
Small pieces of organs were fixed in Palade's solution(Palade phosphate buffer sol. pH. 7.2 added one per cent osmic acid) for 2 hours at 4。C., dehydrated, embedded in metacrylated at 50。C over night and sectioned 20-30mμ with ultramicrotom(Eitachi UM-6).
Carbon-coated thin sections were prepared in vaccum evaporator and examined under the electronmicroscope(Hitachi Hs-6).
The morphological changes of Clonorchis sinensis after the Hetol administration were observed under light microscope and electron microscope.
A. Light microscopic observation:
In the group of 25mg/kg, the size of the worms reduced 1/2 t 1/3 at 24 hours after the administration compared with those of the control group, but the morphological changes were mild in testis and ovary.
In group of 50mg/kg, partial necrosis of testis was observed about 30 per cent 48 hours after the drug administration.
In group of 200mg/kg, the administration, the degenerative changes were remarkable in testis, ovary and excretory bladder, after 72 hours of the drug administration. Genital organs had completely disappeared in 10 per cent of the worms.
In the group of 500mg/kg, most of the bile ducts contained no worms at 72 hours after the administration.
The egg out-put increased in the group of 100mg/kg after 24 hours of the drug administration and the uterine wall also showed morphological change.
Swelling of excretory bladder was observed in the 72 hours group given 200mg/kg of Hetol and the 48 hours group given 200mg/kg of Hetol.
B. Electronmicroscopic observation:
The cristae of the mitochondira in testis, ovary and uterus showed degenerative changes.
The mitochondria itself swelled slightly with the dose of 100mg/kg but atrophied in the group 200mg/kg, and disappeared with the dose of 300mg/kg.
The nucleus began to degenerate with the dose of 25mg/kg, and its changes became severe with the dose of 200mg/kg-300mg/kg.
Vacuole-like degeneration of the nucleus was observed with the dose of 300mg/kg
The severity of the changes of organs were in decreasing of testis, ovary, uterus and cuticle.
The morphological changes of mitochondria in intestine, reticular tissues, suckers and vitelline glands of the experimental group were almost the same as in the control group.
The morphological changes of Clonorchis sinesis by the administration of 1,4-Bis-Trichlormethy1-benzol(Hetol) to infested rabbits in various doses are as follows.
The morphological changes of Clonorchis sinensis was paralle to the dose of Hetol and the period after the drug administration.
Changes appeared mainly in sexual organs. The swelling of the cristae of the mitochondria was seen following the dose of 50-100mg/kg.
Mitochondria atrophied after the dose of 200mg/kg, and completely disappeared following 300mg/jg or more.
Consequently it was concluded that the 1,4 Bis-trichlormethy1-benzol effected mainly sexual organs of Clonorchis sinensis, and led to degeneration.