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Studies on the transmissibility of pathogenic-organisms to liver by migrating larvae of liver fluke and hookworm
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Korean J Parasito > Volume 6(1):1968 > Article

Original Article
Korean J Parasitol. 1968 Jun;6(1):35-44. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1968.6.1.35
Copyright © 1968 by The Korean Society for Parasitology
Studies on the transmissibility of pathogenic-organisms to liver by migrating larvae of liver fluke and hookworm
Hong Chong Lim
Department of Parasitology, Yonsei University Medical College, Korea.
Abstract

In order to confirm whether the migrating larvae of parasites could carry pathogenic organisms into liver and cause hepatitis, a series of experiments has been carried out.

A. Clonorchis sinensis
Recovery rate of larvae in the abdominal cavity of rabbits

One to seven days after the administration adolescariae were recovered from the abdominal cavity in less than l percent of the total number of metacercariae given. Generally, 1~6 larvae were found from each animal which was given 900~1,000 metacercariae, though many larvae were already found in the common bile ducts or remained still in intestine.

Fate of Clonorchis sinensis in abdominal cavity

The young or mature worms which were introduced directly into the abdominal cavity were examined l5, 32, 40 and 42 days after the inoculation. Several larvae were found on the surface of liver in four animals. All the worms on the surface of the liver were dead and the biopsied liver tissues on the area where the worms were attached showed no pathological changes. Two of them were between bile duct and liver tissue but pus cell infiltration surrounding them was observed. In every case, pus cell infiltration was found in the peripheral portion of the liver and pus nodules on the surface of intestine and mesentery. The nodule in the intestinal wall contained the eggs of Clonorchis sinensis. Two worms in the abdominal cavity were still alive. From the above results it is suggested that the larvae of Clonorchis sinensis were capable of penetrating the intestinal wall and reaching the organs in the abdominal cavity and surviving for l5-42 days, but they were unable to penetrate the organs. No bacterial flora appeared from the lesion by culture method.

Fate of Clonorchis sinensis which was inoculated into the peripheral region of liver

Small abscess was observed at the same area. Microscopically, the area became edematous and the vessels in the peripheral region were dilated. The parasites became necrotic and amorphous. Pathologically the lesions appeared as eosinophilic masses and neutrophile leukocytes were infiltrated surrounding the masses. In some cases, the dead worms were found apart from the original place of inoculation but no leukocyte infiltration was found. There was linear infiltration between the original site and the portion where the dead worm was found. The distance from the capsule varied from 0 to 4 mm. Sometimes, the eggs of Clonorchis sinensis were also found. In all cases, there were no living worms in liver tissues and hepatic ducts. In all case, the bacteriological examination was negative.

Do Clonorchis sinensis transfer the microorganism?

Five adult worms of Clonorchis sinensis were incubated in the saline solution containing Staphylococcus aureus. The intestinal contents of these worms were cultured in the Nutient-agar plate and examined by Methylene Blue and Gram's stain. The area of liver tissue where the Clonorchis sinensis were inoculated showed no inflammatory changes after the 3 days of inoculation but no living Staphylococcus aureus was found in the culture media with which the pieces of liver tissues were smeared.

Hookworm

Cutaneous infection: Four to eight days after the cutaneous infection of Ancylostoma caninum, the mice were sacrificed. Grossly, there was no abnormal finding in liver. The pieces of liver tissues were smeared on the Nutrient-agar plate, and cocci were found in four out of six examined. The microorganism were confirmed as the same species of Diplococcus pneumoniae which were grown in the hookworm culture media.

Oral infection: 1,000 filariform larvae of Ancylostoma caninum were given orally. 24 hours later, the mice were sacrificed and the pieces of liver tissue were smeared on the Nutrient-agar plate. After 50 hours at 36 C, the bacterial colonies were examined bacteriologically. Staphylococcus albus was found from two out of four samples. Grossly there was no abnormality on the surface of liver, but microscopically there were spots like microabscesses which were infiltrated by leukocytes. The larvae were also found from other portions of liver tissues and they were surrounded by yellow colored material.

In another experiment, a combination of Ancylostoma duodenale and Staphylococcus aureus was fed to mice. The mice sacrificed five days after the oral administration of Ancylostoma duodenale cultivated in the media containing Staphylococcus aureus.

The liver pieces were examined routinely. The larvae cultivated in normal tap water which contained no Staphylococcus aureus was used as control. In the experimental mouse, the cocci appeared in the liver. Pathologically, microabscesses infiltrated with neutrophile leukocytes were found, but there was no manifestation of inflammatory change due to Staphylococcus aureus. There was only mechanical trauma due to the larvae penetration. Haemorrhage appeared only where the larvae were found.

Figures


Fig. 1
Purulent nodule attached to mesentry of rabbit abdominal cavity, in which egg of Clonorchis sinensis was found.


Fig. 2
Amorphous eosinophilic masses (EM) formed by necrosis of Clonorchis sinensis worm n rabbit liver. H & E stain, × 40.


Fig. 3
Linear cell infiltration caused by migration of Clonorchis sinensis in rabbit liver. H & E stain, × 40.


Fig. 4
Clonorchis sinensis in rabbit liver capsule. H & E stain, × 40.


Fig. 5
Mouse liver lesion caused by infection with Toxocara canis cultured in the medial contaminated with Staphylococcus albus. Larva was surrounded by the materials stained as yellow color. H & E stain, × 40.


Fig. 6
Mouse liver lesion caused by infection with Ancylostoma duodenale cultured in the medial contaminated with Staphylococcus albus. Bleeding by perforation by larva was seen. H & E stain, × 40.

Tables


Table 1
Recovery rate of Clonorchis sinensis larvae from rabbit abodominal cavity, orally infected with metacercariae per os.

× Not detected

+ Number of larvae 21-40

⧺ Number of larvae more than 40



Table 2
Fate of Clonorchis sinensis metacercariae inoculated into abodminal cavity of rabbit.

*From No. 3 rabbit, one small nodule including ova was recovered from the intestinal wall of rabbit.



Table 3
Bacteriological test of intestinal contents of Clonorchis sinensis culutred in the media contaminated with Staphylococcus aureus.


Table 4
Bacteriological test of mice liver infected with Ancylostoma caninum cultured in the media contaminated with Diplococcus pneumoniae.


Table 5
Bacteriological test of mice liver infected with Ancylostoma caninum cultured in the media contaminated with Staphylococcus aureus.


Table 6
Bacteriological test of mice liver infected with Ancylostoma duodenale cultured in the media contaminated with Staphylococcus aureus.

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