• Parasites, Hosts and Diseases
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• v.50 no.2
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• pp.103-111
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• 2012

Intestinal giant-cystic disease (IGCD) of the Israel carp (Cyprinus carpio nudus) has been recognized as one of the most serious diseases afflicting inland farmed fish in the Republic of Korea, and Thelohanellus kitauei has been identified as the causative agent of the disease. Until now, studies concerning IGCD caused by T. kitauei in the Israel carp have been limited to morphological and histopathological examinations. However, these types of diagnostic examinations are relatively time-consuming, and the infection frequently cannot be detected in its early stages. In this study, we cloned the full-length 18S rRNA gene of T. kitauei isolated from diseased Israel carps, and carried out molecular identification by comparing the sequence with those of other myxosporeans. Moreover, conventional PCR and real-time quantitative PCR (qPCR) using oligonucleotide primers for the amplification of 18S rRNA gene fragment were established for further use as methods for rapid diagnosis of IGCD. Our results demonstrated that both the conventional PCR and real-time quantitative PCR systems applied herein are effective for rapid detection of T. kitauei spores in fish tissues and environmental water.

• Parasites, Hosts and Diseases
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• v.28 no.3
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• pp.183-194
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• 1990

In an attempt to develop prophylactic and therapeutic measures of the intestinal giant-cystic disease caused by Thelohanellus kitauei in the Israel carp, Cyprinus carpio nodus, pathological observations were conducted upon the carps which were hatched in May 1988 and raised in a net cage fish farm at the Soyang lake, managed by Horim Fisheries for the period of 21 months with 1~2 months interval. After a gross inspection of the carps, necropsy was carried out periodically in order to clarify the pathological changes in various internal organs and muscular tissues. Also. the prevalence of the disease was checked during the period from 1988 to 1990. Gross inspections revealed that the infected carps showed some degree of fading in body and gill color, back-emaciation symptoms, reddish anus accompanying erosion and relaxation and pot-belly, as well as discharge of yellowish white mucoid material from the anus. However, most carps died eventually of intestinal obstruction. Other significant necropsy fadings included cyst formation of various size in the intestinal mucosa, ascites, anemic condition through internal organs and muscular tissues, hyperemia and dilation of intestines with decreased tension, thinness and fragility, and full contents of semi-fluid or yellowish white mucoid material in the intestinal canals. Based on the morphological characteristics of the spores found in the cysts, parasitic location in the intestines, macro- and microscopic findings of the lesions, the parasites were identised as Thelohanellus kitauei Egusa os Nakajima, 1981. Although monthly changes of water temperature were distinct, the extrusion rates of the polar filaments of the spores stayed constant throughout the year with an exception of a lower rate in July, The lesions initiated from mucosa and submucosa in early July became large swellings and then complete mature (orms following the peracute course. From late August the upper cysts were gradually opened and most of the spores were dispersed from anus into the surrounding water through December but only a few lasted until next April. The cysts were completely recovered until next September. Comparing the incidence and prevalence of the disease by year tremendous infection and death rates were checked in the first prevalent year, 1988, but the rates were significantly decreased in the second year, and showed an almost normal status in the third year, 1990. As the above summarized results showed, the disease entity might come to an end in three years after the first prevalent year, however, the spores must be strictly prevented because they could be infective in the water for one year.

• Journal of fish pathology
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• v.8 no.1
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• pp.13-21
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• 1995

Four species of Actinosporeans, Aurantiactinomyxon sp. A, B, C and Neoctinomyxon sp. D were released from Oligochaete, Branchiura sowerbyi captured from three reserviors, where intestinal giant-cystic disease in carp had occured from June to September, 1994. All part of the intestinal epithelial tissue from the gullet to the anus of B. sowerbyi were infected by Actino-sporeans, and many mature Actinosporean were seen more easily at the posterior parts of the body. Just before releasing, mature Actinosporean sporozoites were divied into each individual from the intestinal epithelial tissue of Oligochaete, while immature ones had 6 spores ($20{\times}25{\mu}m$ in size) per each in the oocyst ($60{\times}65{\mu}m$ in size). A total of 1, 762 of B. sowerbyi were investigated in three reservoirs, 86 individuals (4.88%) of them were infected; 0. 74% (13 ind.) of Aurantiactinomyxon sp. A, 2. 27% (40 ind.) of Aurantiactinomyxon sp. B, 1. 59% (28 ind.) of Aurantiactinomyxon sp. C, and 0. 28% (5 ind.) of Neoactinomyxon sp. D. At the room temperature of 22.6-$30.7^{\circ}C$, number of extrusion dates of Actinosporeans from B. sowerbyi for 32 days are 1 day (23.3% of total, 1 time) or 5 days (11.7%. 5 times), and the majority was finished within 15 days, however, 6.7% of total were released for 32 days.

• Parasites, Hosts and Diseases
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• v.28 no.4
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• pp.241-252
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• 1990

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.

• Parasites, Hosts and Diseases
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• v.31 no.1
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• pp.57-66
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• 1993

The potential of fumagillin dicyclohexylamine salt to treat and prevent intestinal giant-cystic disease in Israel carp, Cyprinus carpio nudus, was monitored in field experimental studies. In experiment 1 (therapeutic), most fish were already naturally infected with more advanced stage of Relohqnellu: kitnuet. Fumagillin was administered to ash (mean body weiht of 830 g for a Penod of one month at a dose of 10.62 mg in the first group and 5.3 mg in the second group per fi sh per day. In experiment 2 (fprophylactlcl), most flesh also were already naturally infected with an early developmental stage of the protozoa and fish (average body weight of 484 g) were administered fumagillln for 45 days at a dose of 3.95 mg per fish per day. In both experiments, the cumulative mortalities of fish and the extrusion rates of the polar filaments of the spores were significantly decreased in a dose-independent fashion. In experiment 2 no dead fish were observed. No adverse side effects of the drug were observed among fish from any dosage group. In experiment 2, an oval or dot-like concave lesion of most cysts developed at the 7th day and the vegetative form was never observed at the 17th day postmedication and the cysts were grossly reduced in size as compared with the control group, beginning at the 24th day until the end of the study. In contrast, it was scarcely effective to the cysts in experiment 1. Taking the seasonal development of the protozoa into consideration, the above results revealed that oral administration of fumagillin at 3.95 mg/500 g body weight/day for a month Is the optimal dose for the treatment and prevention of thelohanellosls caused by T kitnuei among C. carpio nudus.