This study was performed to observe the chronological changes in the tegumental structure of M. yokogawai using scanning electron microscope. The subjected worms were encysted metacercariae obtained from the sweetish, and 2-day, 1-week and 4-week old worms experimentally reared in albino rats. The results are as follows: 1. The tegument of encysted metacercariae showed many transverse shallow rugae, which were more remarkable in posterior half body, i.e., posterior to ventral sucker. The whole surface was armed with many scale-like spines; 7~8 pointed ones on anterior body and 2~3 pointed on posterior body. The ciliated knob-like papillae (Type I) were abundant around oral and ventral suckers, which grouped 2, 3 or 4 in number in most cases. A few round swellings of tegument(Type lI were observed only on oral sucker. 2. The tegumental surface of 2-day old worms showed deeper rugae, and the anterior half covered with knob-like processes of distal cytoplasm and the posterior half with cobblestone-like ones. Interspinous space became more wide and 9 pointed spines appeared on anterior dorsal surface. The sensory papillae enlarged but not changed in their distribution. 3. The tegument of 1-week old worms revealed knob-like cytoplasmic processes in posterior half body and velvety ones around oral sucker. The scale-like spines of anterior half body changed remarkably to the slender ones of posterior body at the level of ventral sucker. In dorsal surface, the arrangement of the Type I papillae were bilaterally symmetrical. 4. The tegument of 4-week old worms were finely differentiated and the posterior tegument covered with velvety cytoplasmic processes. The spines had remarkably grown in length and width but the density remained nearly unchanged. The papillae also became larger but their morphology and distribution were not different from younger worms. However, the round elevation of cytoplasmic ridges (Type III papilla) appeared bilaterally on inner wall of oral sucker, approximately 8 in number. From the above results, it is considered that the tegument of juvenile iH. yokogawai continued to differentiate until 4 weeks after infection.
The in situ posture of anterior body of Metagonimus yokogawai was observed in experimental metagonimiasis of dog. The metacercariae were collected from naturally infected sweetfish by peptic digestion; a total of 8 dogs was orally infected with 10,000 metacercariae respectively. Two dogs were killed on 3 days, 9 days, 4 weeks and 10 weeks after the infection. The postures of worms in histological section of small intestine and of whole worms collected from the fixed intestinal mucosa were examined by light and scanning electron microscope. The results were summarized as follows: 1. The recovery rates of worms were 42.6% on 3 days, 55.0% on 9 days, 33.2% on 4 weeks and 9. 8% on 10 weeks after the infection respectively. 2. In histological sections of small intestine, most of worms were found at intervillous spaces as ovoid sections of posterior body. However, many worms, especially in 3 day-old worms, revealed protruded anterior body in glandular lumens of crypt. Some sections of anterior body were bifurcated or sacculated. 3. The worms collected from fixed intestinal mucosa under dissecting microscope exhibited a variety of postures. Many worms showed flat shapes with a concavity or curvatures. However, in many worms, the anterior body made a single or multiple protrusions. 4. By SEM observation of protruded anterior body, a longitudinal groove was found. 5. The frequency of worms with protruded anterior body decreased in 9 days, 4 weeks and 10 weeks than in 3 day-old worms. The above findings indicated that the anterior body of juvenile and adult M. yokogawai protruded to lumens of glandular crypt by folding their lateral portions to make a reversible tubelike structure. Frequent multiple protrusions were considered to be made to adapt the microniche of glandular crypts of dog intestine.
This study was performed to observe histopathological changes and serological reactions in chronic anisakiasis of rabbits. Each rabbit was infected per os with 30 larvae of Anisakis type I. Their sera were collected chronologically and the rabbits were killed for histopathological examination, 3, 13, 20, 30, 60, 90 and 150 days after the infection. The results were summarized as below. 1. Most of the larvae were recovered from the stomach, but a few from the omentum, intestine, mesentery and abdominal wall. The recovery rates and distribution of worms by organ were not differed by duration of infection. 2. Histologically the lesion was abscess type on 13 days, i.e., the dead worms were surrounded by fibrinous exudate, histiocytes and thick zone of numerous inflammatory cells. After 30 days, histiocytes were found to invade the worms and the lesion was changing into abscessgranulomatous type. Also a calcified worm was found on the 30th day. After then the worms were observed to be dissolved slowly until 90 days. On 150 day, only one calcified worm was observed. 3. The levels of serum IgG antibody by ELISA reached their maximum 30 days after the infection. After then, it decreased slowly until 150 days after the infection. Above serological and histopathological findings indicated that antigenic stimulation from degenerating Anisakis larvae was the greatest during the first 30 days after infection. This period was corresponding with the beginning of worm resolution or calcification. Serologic test by ELISA would be a valuable tool for confirming chronic anisakiasis.
The growth and development of the metacercariae of F. seoulensis cultivated in vitro or on the chick chorioallantois were assessed by comparison with the optimum process of maturation in albino rats and new born chickens. The process of maturation was divided for convenience into six stages: Stage 1 ; cell multiplication, Stage 2; body shaping, Stage 3; separation of genital anlagen, Stage :1 organogeny, Stage 5; gametogony, and Stage 6: oviposition. In Hank's and Tyrode's .solutions, the metacercariae were alive up to 200 days or more at $4^{\circ}C$ without any development. The in vivo maturation process in rats or chicks was as follows: stage 1 from 6 hours; stage 2 from 24 hours; stage 3 from 48 to 72 hours; stage 4 from 3 to 4 days; stage 5 from 4 to 5 days; and stage 6 from 5 to 8 days. Despite unsuccessful infection of the metacercariae to 12 day old chicks, fully mature worms of stage 5 or 6 were recovered from new born chicks (1 to 2 days old), The metacercariae of F. seoulensis grown in vitro were up to stage 3 and no further maturation was observed. Of various media employed, the medium NCTC 109 (Gibco) or NCTC 135(Gibco) supplemented with 20% egg yolk or 20% whole egg macerate or 0.5% yeast was basically required for the earlier development of the fluke. It took 16.1 days(in average) to reach the stage 3 after cultivation. The metacercariae cultivated on the chorioallantoic membranes of 6∼13 day old chick embryo at 37∼38℃ showed their full development up to stage 5 or 6. However, the worms were in general remarkably retarded, compared with those grown in rats or chickens. In the experiments of worm transplant, although the transfer was failed from in vitro culture to in vivo of rats(Per os), the transplants from in vitro culture to the chorioallantois and from the choriollantois to in vivo of rat host were successful with or without development of the transferred worms. In the present study, it was observed that the metacercariae of F, seoulensis can be maintained in vitro media with poor development as well as fully matured in 1 to 2 day-old chicks or on the chorioallantois at a very low rate.
In a basic attempt to develop the prophylactic and therapeutic measures on intestinal giantcystic disease of the Israel carp, C), prinks carpio nudum, the effects of physical and chemical factors on viability or survival of the spores of Thelchcnellus kiteuei were checked in vitro by means of extrusion test on the polar filament. When the fresh spores suspended with 0.45% and 0.9% scdium chloride solution and distilled water were laid at $5^{\circ}C$ and $28^{\circ}C$ for short terms, the extrusion rates increased until the 3rd day, meanwhile when son;e of them were suspended with Tyrode's solution at $-70^{\circ}C$ the rates increased gradually until the 8th day. Viabilities of the spores suspended with 0.9% saline and added antibiotics to the suspension at $5^{\circ}C$ for long terms lasted for 997 days and 1, 256 days (presumed values) at maximum, respectively. The spores suspended with distilled water at $28^{\circ}C$ for long terms survived 152.4 days, but the spores suspended with Tyrode's solution at $-70^{\circ}C$ for long terms showed almost the same viable pattern as early freezing stages up to 780 days. The spores suspended with Tyrode's solution, frozen at $-70^{\circ}C$ and thawed at $5^{\circ}C$, showed the highest rate of extrusion of the polar filament. In the case of frozen spores, the extrusion rates during heating tend to become higher in accordance with the increase of frozen period, and the critical points of 180 day-frozen spores to be killed were generally 78.5 hr. at $60^{\circ}C$, 23.4 hr. at $70^{\circ}C$, 189.1 min. at $80^{\circ}C$ or 10.5 min. at $90^{\circ}C$. The longer the spores were frozen, the more time was needed for the death of spores after thawing; 20 days-17.4 days, 100 days-33.2 days, and 400 days-37.8 days. The longer the spores were frozen, the more time was needed for the death of spores at a conventional when they were dried air drying condition, 540 days-23.5 days, 160 days-21.0 days, and 20 days-14.4 days. On the other hand, the longer the spores were frozen, the more spores were dead rapidly when they were irradiated with 10W UV-ray; 100 days-26.0 hr, 300 days-21.9 hr, and 540 days-13.9 hr. The time needed for killing 200 days-frozen spores by various disinfectants at 1, 000 ppd was 5.2 min. by calcium oxide, 10.4 min. by potassium permanganate, 27.8 min. by malachite green and 14.3 hr. by formalin. Transient inhibitory effects of the extrusion of the polar filament were observed by various antiprotozoal and antifungal agents in the descending order of ketoconazole. metronidasole and dapsone. The above results presume that full drying, followed by spraying CaO and maintaining sunny condition for a few days on the concrete bottoms of knish farm may be an effective method for the prevention of intestinal giant.cystic disease.
Intestinal histopathological changes due to infection with Echinostcma hortense (Trematoda) were studied in rats after experimental infection with the metacercariae. The metacercariae were obtained from the tadpoles of Rana nigrcmaculata, a second intermediate host infected in the laboratory. Total 18 albino rats(Sprague-Dawley) were given 200 matacercariae each and sacrificed on the day 1, 3, 7, 11, 22 or 44 post-infection(PI) Segments of- the small intestine at 1, 3, 5, 8 and 30 cm posterior to the pylorus(PTP) were rejected and studied histopathologically. 1. The flukes were seen to have intruded into the intervillous space in the upper small intestine at early stages(1∼3 days PI), however, they were located mainly in the intestinal lumen at later stages(7∼44 days PI) . The flukes were sucking and destroying the epithelial layers of villi with their oral and ventral suckers. 2. Histopathological changes of the intestine were recognizable in as early as 1∼3 days after infection, and the changes became severer as the infection progressed. 3. The intestinal mucosa was histopathologically characterized by villous atrophy and crypt hyperplasia throughout the infection period. Major villous changes were blunting, fusion, severe destruction and loss of epithelial layers of villi. Villous/crypt(V/C) height ratio was remarkably reduced from 3 : 1 in controls to 1 : 1 in severely infected animals. In the stroma of villi, inaamma- tory cell infiltrations, vascular congestion, edema, and/or fibrosis were recognized. The goblet cells were increased in number after 11 days PI. It was revealed in the present study that the pathological changes in the intestine of rats infected with E. hortense were chieay confined to the mucosal layer of the upper small intestine, however, the changes were very severe accompanying remarkable destruction of villi and loss of mucosal integrity, and persistent until 44 days PI.
PraziquantEI ($Distocide^{\circledR}$), the KcrEan product, was tEstEd for its safety and Efficacy in treatmEnt of Clonorchis sinensiJ infccticn during the period from April to SeptembEr, 1983 in Korea. A total of 55 egg positive cases were selected and treated with the regimen of 25 mg/kg t.i.d. for 1 day (total 75 mg/kg). The follow-up stool examination was done in 47 cases by cellophane thir;k smear and Stoll's egg counting techniques. The 8 uncured cases were treated again with the same regimen. The laboratory tests for blood picture and liver function were done in 27 cases and compared before and after the treatment. The results obtainEd are as follows: 1. After single course treatment, the cure and egg reduction rates were 83.0 and 99.1% respectively. With the second treatment, excellent results of 100% in both rates were obtained. 2. Several kinds of side effects such as dizziness, headache, etc. were complained by 29 cases (61.7 %), however, those were so mild and transient that no special treatment was necessary. 3. No significant change in laboratory findings was recognizable before and after the treatment. From the above results, it is concluded that $Distocide^{\circledR}$ is as effective and safe as $Biltricide^{\circledR}$ and highly recommendable in treatment of C. sinensis infection.
A pathogenic free-living amoeba, Naegleria fowleri, is a causative protozoan parasite of primary amebic meningoencephalitis in human and experimental animals. It is known that humoral and cellular immunity contribute as the defence mechanism of host against this organism. Recently splenectomy has been argued on its effect on host defence mechanisms. The present study was aimed to observe the enact of immunization in splenectomized mice. For immunization, $5~10{\times}10^5$ trophozoites of Naegleria fewleri o 359 were intraperitoneally inoculated once a week for two weeks to BALB/c mice, and $5~10{\times}10^4$ of ameba trophozoites were intranasally inoculated for infection after splenectomy and/or immunization. ELISA technique was applied for the detection of seum IgG antibody levels. Experimental animals were divided into 4 groups; I. splenectomized and immuniEed; ll. splenectomized only; III. immunized only; IV. not splenectomized nor immunized. The results obtained were as follows: 1. Mortality rates of splenectomized and immunized mice in group I (38.1%) and immunized only in group III (25.0%) were lower than those of not immunized mice in group II (50%) and control group, IV (46.4%). 2. Survival times of mice in group I, II, III and IV were $20.1{\pm}3.6$, $17.3{\pm}4.5$, $20.4{\pm}7.0$ and $19.6{\pm}7.6$ days respectively, and there were no significant differences between them. 3. ELISA values (absorbance at 492nm) of group I (1, $10{\pm}0.29$) and group III ($1.31{\pm}0.28$) were significantly higher than that of group IV($0.24{\pm}0.37$) at day 31 of infection (p<0.05). Conclusively, it is presumed that humoral immunity against N. fowleri may operate as ever, after immunization, even though the mouse was splenectomized.
In order to observe the growth and development of Fibricola seoulensis metacercariae, the tadpoles of Rana nigromaculata were experimentally infected with the cercariae. The meta cercariae of various developmental stages were recovered from the tadpoles after 2 to 65 days of infection. They were prepared for morphological observation, and were given orally to mice to observe their infectivity. The following results were obtained. 1. All of the tadpoles exposed to the cercariae were observed to harbour the larvae in their abdominal cavity. 2. The young metacercariae of 2 days after infection were $121.1{\mu}m$ long and $63.3{\mu}m$ wide. They grew linearly for the first 14 days to be $262.0{\mu}m$ long and $166.4{\mu}m$ wide. Thereafter, no more growth recognized until 65 days. 3. The larvae of 2 days old were similar with cercarial body and had 2 suckers, a pharynx, 2 ceca and a primordium of germ cells but no tribocytic organ. On the 8th day, they had tribocytic organ, and their morphology resembled that of mature metacercariae. 4. The metacercariae younger than 10 days could not infect the mice. Only the metacercariae older than 14 days had infectivity. The recovery rates increased by the age of metacercariae from 19.0% in 14 days old to 70.0% in 40 days old. Above findings indicate that the tadpole is indispensable for metacercarial development and it needs at least 2 weeks for maturation. The tadpole is a pivotal host in the life cycle of F. seoulensis for connection between the snail and the frog.
According to the content of sugar and starch of each positional leaf sheath and internode at heading and 4 weeks after it using IR667-Suwon 214 (high yielding var. having tropical Indica parantage) and Jinheung (local leading var. temperate Japonica) rice grown in various cultivation seasons the suitability of grouping into the high sugar type (sugar>starch), sugar, tendency (increasing tendency in sugar content), high starh type (starch>sugar) and starch tendency (increasing tendency in starch) in carbohydrate metabolism was reexamined as follows. 1. Sugar tendency appeared strongly in IR667 than Jinheung, internode than leaf sheath, late season cultivation than early season, 4 weeks after than heading and high temperature than low temperature. Thus at heading, leaf sheath and internode of Jinheung in early and late season cultivation were high starch type, and lower internode in early season cultiattion and leaf sheath and internode in late season for IR667 were high sugar type. In very late season all internodes of both varieties except 1st internode of Jinheung at heading were high starch type. At four week after heading all leaf sheaths except 1st and 4th one of Jinheung and all internodes were high sugar type. High sugar type was intensified 4 weeks after heading in leaf sheaths than in internodes of IR667 in early season and of both varieties in late season. 2. The upper three leaf sheaths and internodes seem to work in the same way for carbohydrate translocation. Among them upper ones showed sugar tendency at heading and starch tendency 4 weeks after heading and it was clear in Jinheung. 3. The later the cultivation season, the higher the carbohydrate content (sugar+starch), and such tendency was clear 4 weeks after heading and in IR667, suggesting teanslocation inhibition by low temperature. 4. Grain filling rate (weight increase per day) was more rapid in early season cultivation and IR667 took shorter days to reach maximum rate. 5. The later the cultivation season, the greater the percent contribution of carbohydrate before heading to yield and it was always greater in IR667, a leaf sheath type. 6. Sugar and starch ratio appears to be determined principally by metabolic characteristics of variety according to growth process and secondly but considerably by environmental factors.
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