We observed the morphological characteristics and identified the species of gnathostome larvae obtained from the imported Chinese loaches. The early third-stage larvae (EL3) were collected from viscera of the loaches and a part of them were infected to mice. The advanced third-stage larvae [AdL3] were recovered from the mice at 4 weeks post-infection, both larval worms were fixed with 10% formalin, cleared in alcohol-glycerin solution, mounted with glycerin-jelly, and observed. A total of 369 EL3 were collected from viscera of 9,493 Chinese loaches. The whole body of EL3 was covered with about 190 transverse rows of minute cuticular spines and 0.624 x 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. AdL3 was 2.660 x 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 Gnathostoma hispidum.
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A scanning electron microscopic study was performed to observe the surface ultrastructures of the third-stage larvae of Gnathostoma hispidum. 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, dryed in critical point dryer, and coated with gold. The specimens were observed with a SEM (DS-130C). 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 micron 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 micron in length, and those in the middle area were about 1.75 micron. 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 Gnathostoma.
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Surface Ultrastructure of the Advanced Third-stage Larvae of Gnathostoma nipponicum E-T. Han, J-H. Lee, S-Y. Choi, J-H. Park, E-H. Shin, J-Y. Chai Journal of Parasitology.2003; 89(6): 1245. CrossRef
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Polymerase chain reaction (PCR) is a powerful technique to detect scanty amount of DNA from living organisms. The present study intended to develop specific primers for PCR diagnosis of pneumocystosis and to evaluate diagnostic efficacy by preparation of template DNAs from invasive BAL fluid and also to screen serum or blood as a non-invasive specimen. Albino rats of Wistar or Fischer strains were experimentally infected by Pneumocystis carinii. Extracted DNAs or cell lysates of their blood, bronchoalveolar lavage fluid, and lung homogenate were used as the template DNA.
Primers were synthetic oligonucleotides among 16s rDNA sequences. All of the primer combinations gave PCR products, but the primer pair of #24 and #27 gave best quality product of 666 bp. The sensitivity of PCR with lysates of BAL fluid was 57.7% but it increased to 84.6% with extracted DNAs.
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Water extract of Coix lacryma seeds (Co-Ex) was separated into several components; dissolved with Tris-Cl buffer and the supernatant (WC1), ammonium sulfate treatment supernatant (WC2) and the pellet (WC3), QAE column chromatography of WC1 and the peak portions; WC4, WC5 and WC6. Murine peritoneal macrophages in DMEM containing 10% heat-inactivated FCS were infected with tachyzoites of Toxoplasma gondii, RH strain, in vitro. By adding modulators such as Co-Ex, WC1,2,3,4,5,6 and LPS or IFN- gamma for 24 hrs, toxoplasmastatic activity of macrophages was examined in relation to nitrite production. Nitrite production of macrophages was enhanced especially in the series of WC2, WC1 and the combination sample (WC1+WC2+WC3) by order, than other components or fractions (WC4, WC5, WC6) tested.
Toxoplasmastatic actions such as percentage of the macrophages infected by T. gondii and fold increase of T.
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Through the results obtained, it is speculated that some components other than the non-proteinous and defatted components in Coix lacryma seeds may contribute to activate macrophages through induction of NO for the biostatic activity.
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