• Korean Journal of Veterinary Research
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• v.38 no.1
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• pp.107-117
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• 1998

The cultivation for development of Ascaris suum from the second-stage larvae($L_2$) embryonated egg and the third-stage of rat-derived larvae($L_3$) recovered from lung of rats were performed to use the screening test of anthelmintics in vitro. The preparations of larvae for cultivation were that the artificially-hatched $L_2$ incubated the embryonated eggs of Ascaris suum in 0.1% formalin solution at $25^{\circ}C$ for 28 days and the rat-derived larvae($L_3$) recovered from the lung of rat infected with the embryonated eggs of Ascaris suum on 7 days after infection(DAI). The cultivation for development of Ascaris suum from the embryonated eggs($L_2$) and the rat-derived larvae($L_3$) for 14 days in RPMI medium 1640(with 5% bovine calf serum) were as follows : 1. The sizes of the liberated larvae($L_2$) which were artificially hatched from embryonated eggs with glass beads(diameter 5mm) were $190{\sim}250{\mu}m$ on 1 days in culture(DIC). The second-stage larvae were molted into third-stage larvae(early $L_3$; $250{\sim}300{\mu}m$) and the features of these larvae were first observed such as cephalic cuticle, esophageal lumen and anus etc. on 5 DIC and the sizes of late third-stage larvae were $250{\sim}450{\mu}m$ on 10 DIC. The sizes of early fourth-stage larvae($L_4$) were $500{\sim}700{\mu}m$ and the features of these larvae were more pronounced in internal organs on 15 DIC. 2. The sizes of third-stage larvae($L_3$) recovered from the lung of rats were $1,340{\sim}1,370{\mu}m$ and the feartures of cephalic cuticle, esophageal lumen, intestine, rectum, anus were visualized by inverted microscope on 1 DIC. The fourth-stage larva($L_4$) completed by third ecdysis were recognizable and sizes of early fourth-stage larvae were developed as $1,400{\sim}2,200{\mu}m$ on 5 DIC. The sizes of middle fourth-stage of larva were $1,900{\sim}2,300{\mu}m$ and the thickened epithelial rectum was observed on 10 DIC. The rectum and anus of late fourth-stage larva($L_4$ $2,500{\sim}3,200{\mu}m$) had developed completely in RPMI medium 1640 on 15 DIC.

• Parasites, Hosts and Diseases
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• v.34 no.3
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• pp.161-168
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• 1996

We observed the morphological characteristics and identified the species of gnathostome larvae obtained from the imported Chinese loaches. The early third-stage larvae ($EL_3$) were collected from viscera of the loaches and a part of them were infected to mice. The advanced third-stage larvae ($AdL_3$) were recovered from the mice at 4 weeks post-infection. both larval worms were fixed loth 10% formalin, cleared in alcohol-glycerin solution, mounted with glycerin-jelly, and observed. A total of 369 $EL_3$ were collected from viscera of 9,493 Chinese loaches. The whole body of $EL_3$ was covered with about 190 transverse rows of minute cuticular spines and $0.624{\;}{\times}{\;}0.101{\;}mm$ in average size. A pair of lips were protruded at the anterior end, and the muscular esophagus and brownish intestine were followed. The characteristic head bulb was provided with 4 rows of hooklets. The average number of hooklets in the respective row was 36.7, 39.5, 41.6 and 44.3 posteriorly $AdL_3$ was $2.660{\;}{\times}{\;}0.346{\;}mm$ in average size, and retained the esophagus (about 0.755 mm length) and cervical sac (about 0.355 mm length). The average number of hooklets in the respective row on the head bulb was 39.0, 41.9, 43.9 and 45.6, posteriorly. On the basis of the morphological characteristics, they were identified as the third-stage larvae of Gnathostomc hispinun.

• Korean Journal of Veterinary Research
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• v.38 no.3
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• pp.589-594
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• 1998

The in vitro screening tests against the in vitro cultivated $L_3$ of Ascaris suum (in vitro $L_3$), which were cultivated from the embryonated egg to third-stage larva on 7 days in culture(DIC) and the in vivo rat's lung-derived $L_3$ of Ascaris suum (in vivo $L_3$), which were recovered from the lungs of rat on 7 days after infection, carried out in order to compare the anthelmintic efficacy of in vitro $L_3$ and that of in vivo $L_3$ in RPMI medium 1640 with 5% bovine calf serum. And also a screening test of efficacy against adult worms of Trichuris suis performed. The efficacies of screening tests were as follows : 1. The screening efficacies of abamectin and ivermectin against the in vitro $L_3$ were all 100% at the 10ppm concentration in RPMI medium 1640 on 5 DIC. 2. The screening efficacies of abamectin and ivermectin against the in vivo $L_3$ were all 100% at the 20ppm on 5 DIC or at 40ppm on 3 DIC. 3. The screening efficacies of abamectin and ivermectin against the adult worms of Trichuris suis were all 100% at 20ppm on 4 DIC. And therefore, the in vitro cultivated $L_3$ of Ascaris suum were used in the screening test as well as the in vivo rat's lung-derived $L_3$ of Ascaris suum. And also the adult worms such as Trichuris suis and filaroids which is small size and difficult to cultivate to vitro, were used in the screening test in vitro.

• Parasites, Hosts and Diseases
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• v.49 no.1
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• pp.39-44
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• 2011

The infection status of marine fish and cephalopods with Anisakis simplex third stage larva (L3) was studied over a period of 1 year. A total of 2,537 specimens, which consisted of 40 species of fish and 3 species of cephalopods, were purchased from the Cooperative Fish Market in Busan, Korea, from August 2006 to July 2007. They were examined for A. simplex L3 from the whole body cavity, viscera, and muscles. A. simplex L3 were confirmed by light microscopy. The overall infection rate reached 34.3%, and average 17.1 larvae were parasitized per infected fish. Fish that recorded the highest infection rate was Lophiomus setigerus (100%), followed by Liparis tessellates (90%), Pleurogrammus azonus (90%), and Scomber japonicus (88.7%). The intensity of infection was the highest in Gadus macrocephalus (117.7 larvae per fish), followed by S. japonicus (103.9 larvae) and L. setigerus (54.2 larvae). Although abundance of A. simplex L3 was not seasonal in most of the fish species, 10 of the 16 selected species showed the highest abundance in February and April. A positive correlation between the intensity of L3 infection and the fish length was obvious in S. japonicus and G. macrocephalus. It was likely that A. simplex L3 are more frequently infected during the spring season in some species of fish. Our study revealed that eating raw or undercooked fish or cephalopods could still be a source of human infection with A. simplex L3 in Korea.

• Parasites, Hosts and Diseases
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• v.34 no.3
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• pp.169-176
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• 1996

A scanning electron microscopic study was performed to observe the surface ultrastructures of the third-stage larvae of Gnathostoma hispidun. The early third-stage larvae (EL3) were collected from the viscera of Chinese loaches by the artificial digestion method . The advanced third-stage larvae (AdL3) were recovered from mice experimentally infected with EL3. Both larval worms were fixed with 2.5% glutaraldehyde, dehydrated in graded alcohol. dryad in critical point dryer, and coated with gold. The specimens were observed with a SEM (DS- l30C). On the head bulb of both larval stage, the mouth had a pair of lateral lips of equal size and of half moon shape. Each lip had a couple of labial papillae and a small amphid located between the two papillae. The hooklets on the head bulb had single-pointed tips and curved posteriorly. The cuticular spines of EL3 were larger and more densely distributed in the anterior area (about 1.8 Mm in length) and gradually decreased in size and number posteriorly. The cuticular spines in the anterior area of AdL3 were sharp-pointed and about 4.5 Mm in length, and those in the middle area were about 1.75 Mm. The velvety cuticular folds and dot-like cuticular spines were distributed in the posterior area. A cervical papilla was located between the 7th and 8th transverse striations. A dome-like body papilla was located at the posterior 1/4 of body. An ellipsoidal excretory pore was located between the 17th and 18th striations. From the above results, it is suggested that the characteristic SEM findings obtained from this study may be helpful on the species identification of larval Gncthostomn.

• The Korean Journal of Ecology
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• v.8 no.4
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• pp.201-204
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• 1985

The author reared the bandwinged grasshoppers, Oedaleus infernalis infernalis in the growth cabinet controlled as temperature of 25。C and 30。C under the conditiion of photoperiod 16L:8D, light intensity 510$\pm$240lux, relative humidity 65$\pm$3%. The results are summarized as follows: The female has six instars and male five instars. The larva reared in the temperature of 25。C died at the second larva stage, and the development to the third instar was impossible. The developmental periods of the egg stage of both sexes in the 30。C are 24.06 days. In the larval development periods, the fourth statge (19.73 days) was longer than that of any stage, and the second stage has the shortest period (9.9 days) in female. In male, the developmental period (9.59 days). The total developmental periods of female and male from the egg to the fifth instar were approximately 95.02 and 95.04 days, respectively. The total developmental period of female was as long as the period (15.75 days) of sixth instar which is not in the male. The survivorship curve in the temperature of 30。C shows concave type.

• Parasites, Hosts and Diseases
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• v.49 no.1
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• pp.51-57
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• 2011

Four cases of gastric or intestinal myiasis are reported. The cases contain 2 males (1 child 10 years old, and 1 adult 40 years old) and 2 females (1 girl 18 years old, and 1 adult 50 years old) from Minia Governorate, Southern Egypt. Three of them, including cases no. 1, 3, and 4, were gastric myiasis, and complained of offensive hematemesis of bright red blood. Minute moving worms, larvae of the fly, were found in the vomitus. On the other hand, case no. 2 had intestinal myiasis, and complained of abdominal distention, nausea, vomiting, and diarrhea. The stool of case 2 was mixed with blood, and minute moving worms were observed in the stool. Endoscopy was performed to explore any pathological changes in the stomach of the patients. The larvae were collected and studied macroscopically, microscopically, and using a scanning electron microscope (SEM) to identify their species. Three different types of larvae were identified. The larvae isolated from case 1 were diagnosed as the second stage larvae of Sarcophaga species, and the larvae isolated from case 2 were the third stage larvae of Sarcophaga species. On the other hand, the larvae isolated from cases 3 and 4 were diagnosed as the third stage larvae of Oestrus species.

• Parasites, Hosts and Diseases
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• v.58 no.6
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• pp.695-699
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• 2020

Present study was performed to know the infection status of Gnathostoma sp. larvae in swamp eels from Cambodia. We purchased total 30 Asian swamp eels, Monopterus albus, from local markets in Pursat and Takeo Provinces and Phnom Penh on May and November 2017 and May 2018. All collected eels were transferred to our laboratory with ice and each of them was examined by artificial digestion method. A total of 15 larval gnathostomes (1-5 larvae) were detected from 55.6% (5/9) swamp eels in Pursat Province. No larval gnathostomes were found in 21 swamp eels in Takeo Province and Phnom Penh. The advanced third-stage larvae (AdL3) detected were 2.575-3.825 (3.250) mm in length and 0.375-0.425 (0.386) mm in width. They had the characteristic head bulb (av. 0.104×0.218 mm) with 4 rows of hooklets, long muscular esophagus (1.048 mm), and 2 pairs of cervical sacs (0.615 mm). The number of hooklets in 4 rows on the head bulb was 41, 44, 47, and 50. In scanning electron microscopy, characteristic features were 4 rows of hooklets on the head bulb, cervical papillae, tegumental spines regularly arranged in transverse striations, and anus. The larval gnathostomes were identified as AdL3 of Gnathostoma spinigerum based on the morphological characters. By the present study, it has been confirmed that G. spinigerum larvae are infected in Asian swamp eels, M. albus, in Pursat Province, Cambodia.

• Parasites, Hosts and Diseases
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• v.53 no.5
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• pp.619-625
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• 2015

The present study was performed to determine the infection status of swamp eels with Gnathostoma sp. larvae in Myanmar. We purchased total 37 Asian swamp eels, Monopterus albus, from a local market in Yangon in June and December 2013 and 2014. All collected eels were transferred with ice to our laboratory and each of them was examined by the artificial digestion technique. A total of 401 larval gnathostomes (1-96 larvae/eel) were detected in 33 (89.2%) swamp eels. Most of the larvae (n=383; 95.5%) were found in the muscle. The remaining 18 larvae were detected in the viscera. The advanced third-stage larvae ($AdL_3$) were 2.3-4.4 mm long and 0.25-0.425 mm wide. The characteristic head bulb ($0.093{\times}0.221mm$ in average size) with 4 rows of hooklets, muscular long esophagus (1.025 mm), and 2 pairs of cervical sacs (0.574 mm) were observed by light microscopy. The average number of hooklets in the 1st, 2nd, 3rd, and 4th rows was 41, 45, 48, and 51, respectively. As scanning electron microscopic findings, the characteristic 4-5 rows of hooklets on the head bulb, a cervical papilla, tegumental spines regularly arranged in the transverse striations, and an anus were well observed. Based on these morphological characters, they were identified as the AdL3 of Gnathostoma spinigerum. By the present study, it has been confirmed for the first time that Asian swamp eels, M. albus, from Yangon, Myanmar are heavily infected with G. spinigerum larvae.

• Parasites, Hosts and Diseases
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• v.36 no.1
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• pp.15-22
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• 1998

An in uitro culture technique was established for harvesting Strongwloides venezuelensis free-living infective larvae using a nutrient broth medium as a substitute for rat-feces in polyvinyl culture bags ($10{\;}{\times}{\;}12{\;}cm$). The egg hatch rate V) in sterile saline at different incubation temperatures (X) was expressed as the quadratic function, Y = $-0.192X^2$ + 8.673x - 19.550 (r = 0.901). The highest (100%) egghatch rate was observed at $25^{\circ}C$. A significant difference (p<0.05) in development rate W) of free-living infective larvae was observed between different concentrations of nutrient broth (X) which was highest (20.6%1 in 0.12% nutrient broth concentrations, incubated at $20^{\circ}C$ for 5 days [Y = $-864.032X^2$ + 245.995X- 0.560 (r = 0.875)]. Yields (Y) of infective larvae were observed relatively high when the culture medium was incubated at higher temperatures (X) which peaked at $25^{\circ}C$ (20.0%) than at lower temperatures. $15^{\circ}C$M (10.9%) and $20^{\circ}C$ (18.1%) [Y = $-0.189X^2$ + 8.387x- 72.795 (r = 0.981)]. The period W) required for the development of infective larvae decreased with higher incubation temperatures (X) [Y = $0.035X^2$ - 2.025X + 32.375 (r = 0.995)] The highest yield (19.2%) of infective larvae was obtained from culture bag inoculated with 15.000 eggs than with below and over 15,000 eggs in 0.12% nutrient broth and incubated at $25^{\circ}C$ for 4 days. The newly adapted culture method (from egg to third-stage larva) may be useful as a bio-bar/bioassay system for screening new chemical products, anthelmintics and pesticides, as well as for parasito immunological studies with Strongwloides species.