• Title/Summary/Keyword: third-stage larva

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In vitro cultivation for development of Ascaris suum from the decoated and embryonated eggs(second-stage larva) (시험관내에서 돼지회충(Ascaris suum) 함자충란(L2)의 인공배양)

  • Jee, Cha-ho;Park, Seung-jun
    • 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.

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Identification of larval Gnathostomn obtained from imported Chinese loaches (중국산 수입 미꾸리로부터 획득한 악구충 제3기 유충의 종 동정)

  • 손운목;이순형
    • 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.

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Culture Condition of Entomopathogenic Nematodes Using Galleria mellonella Larva (Galleria mellonella 유충을 이용한 곤충병원성 선충의 배양 조건)

  • 김도완;박선호
    • KSBB Journal
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    • v.13 no.1
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    • pp.31-37
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    • 1998
  • A simple method for the in vivo production of third-stage infective juveniles(IJs) of Steinernema glaseri was developed. Using Galleria mellonella larvae, only IJs can be rapidly generated inadequate quantities for field application. The nematode inoculation concentration and incubation temperature were critically important. The most effective temperature for infectivity of Steinernema glaseri IJs to Galleria mellonella larvae was 33$^\circ C$. However, the total number of menatodes harvested at 25$^\circ C$ about 66,000 IJs per larva was significantly greater than those at other temperatures. The optimal inoculation number of nematodes was 60 to 80 nematodes per host larva. The higher nematode inoculation concentration of 100 IJs per larva caused a rapid decrease in the total number of IJs harvested. As the inoculation medium pH increased, the number of IJs harvested increased and reached about 110,000 IJs per larva at pH 9.0. The pathogenicity of IJs decreased y increasing the salt concentration in the medium.

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Isolation and comparison of somatic and excretory-secretory antigens for serological diagnosis in Trichuris suis (혈청학적 진단을 위한 돼지 편충의 체항원, 배설/분비항원의 분리 및 비교)

  • Jee, Cha-ho;Lee, Chul-soon;Park, Seung-jun
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.159-168
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    • 1999
  • Swine whipworm(Trichuris suis) is cosmopolitan nematode which can cause serious pathology in immature stage(larva2~larva5) of infected pigs, such as anorexia, diarrhea, anemia, and death in heavy infections. In this larval stages, it is very difficult to diagnose the infection of whipworm and to differentiate from other common swine gastrointestinal disorders such as 21 day scours which are associated with TGE virus, rota virus, coccidium, and the stress of weaning. In this experiment, the isolated antigens of Trichuris spp. were carried out to examine the structure and specificity of antigens and to select the reasonable antigens which would be used in serological diagnosis by electrophoresis, Western blotting, ELISA. The results of this experiment were as follows; 1. The common fractions of each Trichuris suis antigen were identified 28,32,45, 80kDa by SDS-PAGE with silver stain and four major fractions could be detected in positive swine sera by Western blot analysis. 2. The OD(optical density) values of somatic and excretory-secretory antigens which were reacted against positive(negative) sera from pigs infected with Trichuris suis by ELISA reader were; 1) OD values($mean{\pm}SD$) of adult somatic antigen against positive(negative) sera were $0.30{\pm}0.12(0.09{\pm}0.006)$ and third-stage larva of somatic antigen were $0.28{\pm}0.038(0.10{\pm}0.005)$. And OD values of excretory-secretory antigens of adult and third-stage larva were $0.24{\pm}0.031(0.11{\pm}0.005)$ and $0.08{\pm}0.013(0.10{\pm}0.003)$, respectively. 2) OD values of adult somatic, larval somatic antigen and adult excretory-secretory antigen response to positive sera were significantly (p<0.01) associated with negative swine sera. And the Cut-off OD values(minimum positive value) were determined to be mean negative value plus 3 SD that would minimized the risk of false positives. 3. The OD values of somatic antigens of T suis and T vulpis against swine positive(negative) sera were $0.30{\pm}0.120(0.09{\pm}0.006)$ and $0.25{\pm}0.141(0.09{\pm}0.003)$. These data mean that the somatic antigens of T suis and T vulpis were able to diagnose T vulpis infection in dogs as well as T suis infection in pigs. These results suggest that somatic antigen of third-stage larva and excretory-secretory antigen of adult T suis could be used the diagnostic antigen by serological test(ELISA) in immature Trichuris spp. infection.

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Bionomical characteristic of Protaetia brevitarsis (흰점박이꽃무지의 생육특성)

  • Kim Ha-Gon;Kang Kyung-Hong
    • Korean journal of applied entomology
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    • v.44 no.2
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    • pp.139-144
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    • 2005
  • This study was performed to investigate the bionomical characteristics of Protaetia brevitarsis in Korea. The imaginal stage was from in early July and the advent of imagoes was the most frequent in early August. Also few imagoes was entered into the hibernation. The larvae inhibited in humus and the period of larva was all completed in late November. At the stage of the third larva, the larvae entered into the hibernation. The average number of laid eggs was 152, and all periods of each stage - eggs, first, second, third instar larvae and pupa - were shortened in $30^{\circ}C$ than in $25^{\circ}C$. As for the longevity of imagoes, female, reared in individual and in low temperature lived longer than male, in group and in high temperature, respectively.

Bionomical characteristic of Allomyrina dichotoma (장수풍뎅이의 생육특성에 관한 연구)

  • Kim, Ha-Gon;Kang, Kyung-Hong
    • Korean journal of applied entomology
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    • v.44 no.3 s.140
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    • pp.207-212
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    • 2005
  • This study was performed to investigate the bionomical characteristics of Allomyrina dichotoma in Korea. The imaginal stage was from late June to late August, and the advent of imagoes was the most frequent in mid-July. The larvae inhibited in humus and the period of larva was all completed in late October. At the stage of the third larva, the larvae entered into the hibernation. The average number of laid eggs was 24, and all periods of each stage - eggs, first, second, third instar larvae and pupa - were shortened in $30^{\circ}C$ than in $25^{\circ}C$. As for the longevity of imagoes, female, reared in individual and in low temperature lived longer than male, in group and in high temperature, respectively.

A case of fatal hyperinfective strongyloidiasis with discovery of autoinfective filariform larvae in sputum

  • KIM Jin;JOO Hyun-Soo;KO Hyang-Mi;NA Min-Sik;HWANG Sun-Ho;IM Jong-Cheol
    • Parasites, Hosts and Diseases
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    • v.43 no.2 s.134
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    • pp.51-55
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    • 2005
  • The autoinfective filariform larva of Strongyloides stercoralis causes hyperinfection in immunosuppressed hosts. Here we report on the case of a male patient who was admitted to the emergency room at Gwangju Veterans Hospital with a complaint of dyspnea, and who was receiving corticosteroid therapy for asthma. Many slender larvae of S. stercoralis with a notched tail were detected in Papanicolaou stained sputum. They measured $269\pm21.2{\mu}m$ in length and $11\pm0.6{\mu}m$ in width. The esophagus extended nearly half of the body length. The larvae were identified putatively as autoinfective third-stage filariform larvae, and their presence was fatal. The autoinfective filariform larva of S. stercoralis has not been previously reported in Korea.

Development of the Band-Winged Grasshoppers, Oedaleus infernalis infernalis at the Constant Temperature (팥중이 ( Oedaleus infernalis infernalis ) 의 恒溫에서의 發育)

  • Joo, Eun Young;Sang Ock Park
    • 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.

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The screening test on the efficacy of anthelmintics by using third-stage larvae and adult of cultivation in vitro (시험관내에서 인공배양한 제 3기 자충 및 성충을 이용한 구충효능 선발시험)

  • Jee, Cha-ho;Park, Seung-jun
    • 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.

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A Case of Anisakidosis Caused by Pseudoterranova decipiens Larva

  • Na, Hak Kyun;Seo, Min;Chai, Jong-Yil;Lee, Eun Kyoung;Jeon, Soung Min
    • Parasites, Hosts and Diseases
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    • v.51 no.1
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    • pp.115-117
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    • 2013
  • Pseudoterranova decipiens larva is a rare cause of anisakiasis. Indeed, prior to the present study, there had been only 12 reports of larval P. decipiens infection in the Republic of Korea. In June 2011, an anisakid larva, 32.1 mm in length and 0.88 mm in width, and finally identified as the third stage larva of P. decipiens owing to the presence of an intestinal cecum but lacking ventricular appendage, was discovered in a 61-year-old woman during the course of endoscopy executed as a part of routine physical examinations. The patient had eaten raw a rockfish 13 hr prior to the endoscopy, but showed no symptoms of anisakiasis. This paper is the 13th report of P. decipiens infection in Korea.