Lee Chan Wuk
Department of Parasitology and Institute of Endemic Diseases, College of' Medicine, Seoul National University, Korea.
Kisaengchunghak Chapchi. 1964 Jun;2(1):63-80. doi: 10.3347/kjp.1964.2.1.63.
Capillaria hepatica is an extremely common parasite of rats. Several human cases have also been reported from various parts of the world and recently these aroused the clinical interests. The present study was undertaken to investigate the biological observations of C. hepatica and the changes occurring in blood picture and serum protein in the experimentally infected hosts. The source of C. hepatica obtained from the deposit of non-embryonated eggs encapsulated in the liver of house rats(Rattus norvegicus) in Seoul. The eggs isolated from the infected liver tissues by the freshly prepared artificial gastric juice at 37 C and embryonated in the incubator 27 degrees to 30 degrees C for four to five weeks. For the observation of migratory pathway to the liver, ten mice were infected orally with 1,000 embryonated eggs of C. hepatica, and another ten mice were infected intraperitoneally. No larvae were found in the washings of peritoneal cavity after oral infection, but after the third day of infection, the larvae were isolated from liver tissues. These indicated that the majority of larvae are transported to the liver by the hepatic portal system. On the other hand, 1,000 embryonated eggs of C. hepatica were inoculated into the peritoneal cavity of mice by mantoux syringe containing antibiotics. One third of inoculated eggs hatched out in the peritoneum during two days after inoculation, hatched in the peritoneal cavity invade directly to the surface of liver. Twenty white rats were infected orally with 1,000 to 2,000 embryonated eggs for the study of the development of C. hepatica in the liver and histopathological changes of the infected liver in the course of infection. C. hepatica in the liver of white rats developed rather slowly at the first tenth day after infection, but at the 13th day developed rapidly in its size. The worms were sexually differentiated at the l7th day after infection. At the 20th fully formed eggs appeared in the white or yellowish lesions on the surface of rat liver and they are also found in uterine tubule of the female worm. After the 33rd day, male worm disappeared and only female worms packed with eggs were detected in the liver tissues. However the long hair-like tightly coiled worms were also usually found in the hepatic cysts, and the degenerated or dead worms were observed in the small cysts on the surface of the liver at the 59 th day after infection. Microscopical examination on the first week after infection revealed inflammatory reactions with the dilatation of central vein, Kupffer cell mobilization, focal necrosis and perivascular infiltration. After two weeks of infection granulomatous inflammation were observed around or adjacent to the worms in the lobules. The worms are surrounded by macrophages, multinucleated giant cells, a dense infiltration of lymphocytes, monocytes, neutrophils and, especially, eosinophils. After the third and fourth week, the microscopical findings of infected rat livers have shown proliferation of connective tissues and regeneration of liver cells. During the fifth to sixth week after infection, rat liver showed marked proliferation of fibrous connective tissues encapsulated the worms and massive deposition of the eggs. At the later time the liver reveals many pseudolobules which are caused by postnecrotic cirrhosis. These are irregularly subdivided into lobule by a fibrous septum. The worms were fragmented by the phagocytes and encapsulated by connective tissues. And then finally they appeared to be replaced by the calcium-like material. The liver shows typical cirrhosis after the eighth week after infection. In order to investigate the changes of blood picture and serum protein components of rabbits infected with C. hepatica, twenty rabbits were divided into four groups by the doses of eggs. Group A was given doses of 1,000 embryonated eggs, group B 5,000 eggs, group C 10,000 eggs and group D 30,000 eggs. The pictures of blood especially leukocyte and eosinophil counts and of serum protein were checked every week for ten weeks in the course of infections. The marked elevation of the leukcocyte, eosinophil counts and percentage of eosinophils was observed at the sixth to the seventh week in the course of infection in all groups of rabbits. At the tenth week after infection a decrease was shown in their counts. However in the heavily infected groups (Group C & D) these values persisted relatively in high levels even thereafter. In the white rats given doses of 1,000 to 2,000 eggs, eosinophil counts increased to the peak at the fourth week and decreased at the seventh week after infection. The changes in serum protein components of infected rabbits were investigated by paper electrophoresis. Blood collections were done by the cardiac puncture in the early morning. Serum total protein was determined by Biurets method, serum protein fractionating and A/G ratio by paper electrophoresis using Whatman No.l filter paper and barbital buffer (pH 8.6, ionic strength 0.06). Total protein increased at the sixth and seventh week after infection and the albumin and A/G ratio had decreased significantly in the heavily infected groups at the fifth and sixth week. The alpha-globulin and beta-globulin were not significant in the lightly infected groups(Group A & B), but they decreased after seventh week in the heavily infected groups. The gamma-globulin and &ggr;/A ratio of the heavily infected groups were significantly increased at fifth to seventh week. Statistically the calculation of entropy was applied to the data obtained in all groups. In the lightly infected groups, the entropy was included almost in the normal ranges, however in the heavily infected groups it was excluded from the normal range during the first to eighth week after infection.
肝毛细线虫是大鼠极为常见的一种寄生虫。世界各地也报告了数例人类感染病例,最近这些病例引起了临床关注。本研究旨在调查肝毛细线虫的生物学特性以及实验感染宿主的血象和血清蛋白变化。肝毛细线虫取自首尔家鼠(褐家鼠)肝脏中包裹的未受精卵沉积物。通过在37℃用新制备的人工胃液从感染的肝脏组织中分离出卵,并在27℃至30℃的培养箱中孵育四至五周使其胚胎化。为观察其向肝脏的迁移途径,给10只小鼠口服1000枚肝毛细线虫胚胎化卵,另10只小鼠腹腔注射。口服感染后在腹腔冲洗液中未发现幼虫,但感染后第三天,从肝脏组织中分离出幼虫。这表明大多数幼虫是通过肝门静脉系统转运到肝脏的。另一方面,用含抗生素的曼氏注射器将1000枚肝毛细线虫胚胎化卵接种到小鼠腹腔。接种后两天内,三分之一的接种卵在腹膜内孵化,在腹腔内孵化的幼虫直接侵入肝脏表面。为研究肝毛细线虫在肝脏中的发育以及感染过程中感染肝脏的组织病理学变化,给20只白鼠口服1000至2000枚胚胎化卵。感染后第10天,白鼠肝脏中的肝毛细线虫发育相当缓慢,但在第13天其大小迅速增长。感染后第17天性别分化。感染后第20天,在大鼠肝脏表面的白色或淡黄色病变中出现完全成熟的卵,在雌虫的子宫小管中也可发现。感染后第33天,雄虫消失,肝脏组织中仅检测到充满卵的雌虫。然而,在肝囊肿中通常也可发现长而毛发状紧密盘绕的虫体,感染后第59天在肝脏表面的小囊肿中观察到退化或死亡的虫体。感染后第一周的显微镜检查显示有炎症反应,中央静脉扩张、库普弗细胞动员、局灶性坏死和血管周围浸润。感染两周后,在小叶中虫体周围或附近观察到肉芽肿性炎症。虫体被巨噬细胞、多核巨细胞、淋巴细胞、单核细胞、中性粒细胞尤其是嗜酸性粒细胞的密集浸润所包围。感染后第三周和第四周,感染大鼠肝脏的显微镜检查结果显示结缔组织增生和肝细胞再生。感染后第五至六周,大鼠肝脏显示出包裹虫体的纤维结缔组织显著增生和卵的大量沉积。后期肝脏出现许多假小叶,这是由坏死后肝硬化引起的。这些假小叶被纤维间隔不规则地细分为小叶。虫体被吞噬细胞破碎并被结缔组织包裹。然后最终它们似乎被类钙物质取代。感染后第八周肝脏呈现典型的肝硬化。为研究感染肝毛细线虫的家兔血象和血清蛋白成分的变化,将20只家兔按卵剂量分为四组。A组给予1000枚胚胎化卵,B组给予5000枚,C组给予10000枚,D组给予30000枚。在感染过程中,每周检查10周的血象,尤其是白细胞和嗜酸性粒细胞计数以及血清蛋白情况。在所有家兔组感染过程中的第六至七周,观察到白细胞、嗜酸性粒细胞计数及嗜酸性粒细胞百分比显著升高。感染后第10周其计数下降。然而,在重度感染组(C组和D组),即使此后这些值仍相对保持在较高水平。给白鼠口服1000至2000枚卵后,嗜酸性粒细胞计数在感染后第四周升至峰值,第七周下降。通过纸电泳研究感染家兔血清蛋白成分的变化。清晨通过心脏穿刺采血。血清总蛋白采用双缩脲法测定,血清蛋白分级及A/G比值采用Whatman No.1滤纸和巴比妥缓冲液(pH 8.6,离子强度0.06)进行纸电泳测定。感染后第六和七周总蛋白增加,重度感染组在第五和六周白蛋白和A/G比值显著下降。轻度感染组(A组和B组)中α球蛋白和β球蛋白无显著变化,但重度感染组在第七周后下降。重度感染组的γ球蛋白和γ/A比值在第五至七周显著增加。对所有组获得的数据进行统计学熵计算。在轻度感染组,熵几乎在正常范围内,然而在重度感染组,感染后第一至八周熵超出正常范围。