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    28 February 1987, Volume 5 Issue 1
    DETERMINATION OF CIRCULATING ANTIGENS IN HUMAN FILARIASIS BY MEANS OF A MODIFIED SANDWICH ENZYME-LINKED IMMUNOSORBENT ASSAY (MS-ELISA) USING MONOCLONAL ANTIBODIES
    1987, 5(1):  1-5. 
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    A MS-ELISA using rabbit anti-microfilarial serum (RAS)-immunoglob ulins as the first antibody and monoclonal antibodies (McAbs) as the second antibody was developed for the determination of circulating antigens in human lymphatic fil ariasis. When McAbs against W. bancrofti microfilarial excretion-secretion antigens (ES-34) and against B. malayi L-3 antigen (HC-11) were used, a deteitability as little as 10 μg/ml and 50 μg/ml were achieved with positive rates of 94.5% (103/109) and 89.0% (97/109) respectively in microfilaremia patient sera (95 bancroftian, 14 brugia n) collected from India and Guizhou, China. A positive correlationship between antigen liters and circulating microfilaria density was observed with ES-34 MS-ELISA. However, in microfilaria-free cases of late stage with hydrocele, chyluria and elephantiasis complications, only 57.4% (31/54) and 61.1% (33/54) were obtained respectively with these two McAbs. Filarial antigens were also detectable in the urine in a part of the micr ofilaremia patients. 85 sera of healthy people collected from non-endemic areas in USA (36) and Guiyang, China (49) showed low positive rates of 0-4.1% with ES-34 and 2 .8-4.1% with HC-11. Negative reactions were observed in sera of all the 30 non-filaria cases harbouring intestinal helminths. The results strongly suggested that the presence of circulating antigens is correlated to the existence of active infections.
    DOT-ELISA USING MONOCLONAL ANTIBODIES FOR IDENTIFICATION OF LEISHMANIA DONOVAN1
    1987, 5(1):  6-8. 
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    Dot-ELISA for monoclonal antibodies specific to different species of leishmania promastigotes from naturally infected sandflies was described. One of the monoclonal antibodies, L9E12 was selected for further analysis. The highest titer of the ascites fluid With promastigotes of Xinjiang strain determined by the Dot-ELISA was 1:655 360. The promastigotes Zi 10 and Zi 11 tested with hybridoma L9E4 ascites fluid showed a titer of 1:81920 dilution which was 3-20 times higher than by the IFA technic.So the Dot-ELISA described is proved to be very sensitive, specific and fast in the determination of promastigotes species from naturally infected sandflies.
    CELL LINES OF HYBRIDOMA SECRETING MONOCLONAL ANTIBODIES AGAINST SCHISTOSOMA JAPONICUM EGG ANTIGENS
    1987, 5(1):  9-12. 
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    After the fusion of SP 2/O myeloma cells with spleen cells of BALB/C mice immunized with SEA of Schistosoma japonicum eggs, 4 cell lines of hybridoma secreting monoclonal antibodies against S. japonicum eggs have been established. Four hybridoma-derived antibodies were measured by IFA with frozen sections of adult worms and 3 different patterns of immunofluorescence reaction were clearly revealed. Of these, 3 monoclonal antibodies presented a high specificity to SEA in ELISA and also showed positive COP and IHA reaction. Another monoclonal antibody was found to be against the common antigenic determinant of Schistosoma, Paragonimus, Clonorchis in I-ELISA, The immunoglobulin subclass of the 4 monoclonal antibodies were identified as IgM.
    EFFECTS OF VARIOUS ANTI-HOOKWORM DRUGS ON THE CHOLINESTERASE ACTIVITY OF NECATOR AMER1CANUS, ANCYLOSTOMA DVODENALE, ANCYLOSTOMA CANINVM AND NIPPOSTRONGYLUS BRAZILIENSIS
    1987, 5(1):  13-17. 
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    This paper presented the cholinesterase (ChE) activity value of Necator americanus (Na), Ancylostoma duodenale (Ad), Ancylostoma caninum (Ac) and Nippostrongylus braziliensis (Nb). The ChE activity of Na was about 40 times higher than that of Ad, The ChE activity of Ad was close to that of Ac while the ChE activity of Nb was approximately one half of that found in Na. Various anti-hookworm drugs including metrifonate, bephenium hydroxynaphthoate, mebendazole, pyrantel pamoate, tetrachloroe-thylene and amidental were given orally to hamsters, puppies and rats infected either with Na, Ad, Ac or Nb to study their effects on the ChE activity of these worms.Metrifonate administered at a single dose of 150mg/kg inhibited completely the ChE activity of adult Na in hamsters as well as that of adult Ad and Ac in puppies. The inhibition of ChE activity by metrifonate in Na was reversible.Bephenium hydroxynaphthoate had no effect on ChE activity of Na in hamsters at a dosage of 3g/kg. However, it inhibited partially the ChE activity of Ad in puppies 24 hours after administration.Pyrantel pamoate, mebendazole, tetrachloroethylene and amidentel had no effect on ChE activity of Na and Ac.
    ANALYSIS ON EPIDEMIC CHARACTERISTICS OF FALCIPARUM MALARIA IN SOUTH HENAN
    1987, 5(1):  18-21. 
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    The epidemic characteristics of falciparum malaria in Xinyang Prefecture of south Henan Province between 31-33°N were analysed basing upon the epidemiological surveys of malaria carried out during the period 1954-84. This area is endemic for vivax and falciparum malaria, the annual malaria parasite rates in local inhabitants varied between 5% and 10%, falciparum malaria or vivax malaria prevailed or coexisted in the past decades. In the 1950's a falciparum malaria outbreak occurred in the west part, since then only two small falciparum malaria outbreaks were reported in the east part in early 1970's. The parasite rate of malaria rose up to 37.9% (P. falciparum .56.7%) in Huzu District of Gushi County in 1974. Since successful malaria control there were only two carriers of P. falciparum in the whole prefecture. However, the parasite rate rose again and it was 24.8% (P. falciparum 79.0%) in Wangliu District of Gushi County in 1984. Recent investigation demonstrated that Anopheles lesteri antkropophagus is the main transmitting vector of P. falciparum and its distribution parallelled with that of P. Jalciparum.
    DIAGNOSING GIARDIASIS BY DETECTING GIARDIA LAMBLIA ANTIGEN IN FECAL MATTER WITH COUNTERIMMUNOELECTROPHOKESIS
    1987, 5(1):  22-24. 
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    Fecal specimens of 35 parasitologically confirmed cases of giardiasis, 41 acute, gastroenteritis, 23 acute baciUary dysentery, 40 normal persons and 15 jirds experimentally infected with G. lamblia were tested with conterimmunoelectrophoresis (CIE). It was found that 33(94%) out of the 35 giardiasis patients and 14(93%) out of 15 infected jirds showed positive reaction, while fecal specimens of other cases, normal persons, or normal jirds all showed negative reaction. Besides, CIE was performed in 4 giardiasis patients before and after metronidazole treatment. Prior to metronidazole, they were all CIE positive, one day post metronidazole, 3 of them were still CIE positive, and Giardia cysts or trophozoites were also present in their stools. However, from the second day onwards, all of them became CIE negative, while cysts or trophozoites also disappeared from their stools. Apparently, detecting Giardia antigen in fecal specimens with CIE of feces is not only a sensitive tool for detecting current infection, but also a, useful tool for evaluating therapeutic effects in giardiasis.
    OBSERVATION ON THE DIFFERENCE IN SUSCEPTIBILITY OF ONCOMELANIA SNAILS TO DIFFERENT ISOLATES OF SCHISTOSOMA JAPONICUM
    1987, 5(1):  25-28. 
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    Two groups of 25 snails each from Sichuan and Yunnan Provinces were experimen-tally infected with miracidia of Schistosoma japonicum from Guichi, Anhui Province and the infection rates were 32% and 28% respectively by histological method. When the same number of snails from Guichi was infected with local isolate of S. japonicum as control, the infection rate was 84%. The speed of development and morphological change of invading larvae of S. japonicum in the two different batches of snails from Sichuan and Yunnan were slower than those in the control snails, indicating a difference in susceptibility.
    IN VITRO EFFECT OF TINIDAZOLE ON TRICHOMONAS VAGINALIS AND TRICHOMONAS HOMINIS
    1987, 5(1):  29-31. 
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    The trichomonadicidal activity of tinidazole (Fasigyn) was evaluated by the ire vitro cultivation of Trichomonas vaginalis and Trichomonas hominis with different drug concentrations. The results showed that, after 48-hour incubation with tinidazole, the 100% lethal concentration was 20μg/ml. The trichomonadicidal effect of tinidazole was one titre lower than that of metronidazole. With regard to T. hominis, after 48-hour incubation with tinidazole, the 100% lethal concentration was 20μg/ml. In comparison with metronidazole, the trichomonadicidal effect of tinidazole was similar.
    INHIBITION OF TEGUMENTAL y-GLUTAMYLTRANSPEPTIDASE OF SCHISTOSOMA JAPONICVM BY PRAZIQUANTEL
    1987, 5(1):  32-34. 
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    γ-glutamyltranspeptidase(γ-GTP) was found to exist in the tegument of S. japonicum, as revealed by Oaks method. Praziquantel at the concentration of 1 × 10-2 mM could inhibit in vitro the tegumental γ-GTP of male worm by 23%. By using intact worms, it was found that the activity of γ-GTP was higher than that of the tegument. The in vitro inhibitory effect of praziquantel on γ-GTP of female schistosomes was higher than that of the male worms. The authors have only measured the in vivo enzymatic activity of male worms, and the significant inhibitory effect of γ-GTP by praziquantel was found. At the same dosage, praziquantel did not show any inhibitory effect on the γ-GTP of the host kidney.
    MEMBRANE STRUCTURE OF 15 SPECIES OF HELMINTHIC EGGS
    1987, 5(1):  35-38. 
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    An investigation by scanning electron microscopy was made on the eggs of parasitic helminths. The eggs were fractured by DMSO freezing etching method. The parasite species examined were Ascaris lumbricoides, Enterobius vermicularis, Gongylonema pul-chrum, Necator americanus, Ancylostama duodenale, Trichuris suis, Clonorchis sinensis,, Paragonimus westermani, Fasciolopsis buski, Fasciola hepatica, Schistosoma japonicum, Taenia solium, Taeniarhyrichus saginatus, Spirometra mansoni, and Macracanthorhynchus. hirudinaceus.The egg shell, protein layer, vitelline membrane and lipid layer varied with the species among nematodes. There is a layer of etching structure on the outer surface of the egg shell in some trematodes and cestodes. According to the thickness of the egg shell, the eggs can be divided into 3 types, i.e. (1) below 0.1 μm, including T. solium egg; (2)0.1-1μm, including the eggs of N. americanus, A. duodenale, E. vermicularis, C. sinensis and S. japonicum; (3)1.1-2 μm including fertilized eggs of A. lumbricoides, eggs of T. suis, G. pulchrum, F. buski, F. hepatica and Spirometra mansoni.The operculum at one end of the eggs in some trematodes and cestodes is a hatching area, through which the larva hatches out. It was found that in some nematodes a characteristic structure at one or both ends of the egg is also present which may also be regarded as the hatching area.The fine tubular system in the embryophore of T. soZiufn and T. saginatus possess; not only microtubes but also micro caverns. In the egg shell of E. vermicularis and S. japonicum, this system is composed of numerous single tubules. In N. americanus egg,, however, only numerous holes are present.
    MECHANISM OF LEVAMISOLE IN PREVENTING HOOKWORM INFECTION
    1987, 5(1):  39-41. 
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    (1) The hookworm larvae which had been immersed in the extract of the mouse's, skin smeared with 0.5% or 1.5% levamisole (LVM) lost the ability of skin penetration; (2) After 1.5% 3H-LVM had been applied onto the belly skin of the mouse the drug radioactivity in the skin decreased rapidly in the first 3 days, but 3 days later, the speed became distinctly slower. After 4-5 weeks, a certain amount of the drug could still be detected. The drug was centered mostly in the applied part of the skin, increased with increasing dosage and was far much higher by cutaneous administration than by oral route. It was suggested that the continued effect of LVM in preventing hookworm larvae infection was due to the drug remaining in the skin for a relatively long time.
    EXPERIMENTAL STUDY ON THE EFFECT OF PLASMODIA ON ERYTHROID COLONY FORMING UNIT (CFU-E)
    1987, 5(1):  42-44. 
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    The dynamic process of the bone marrow and spleen CFU-E from the mice infected with Plasmodium berghei was observed and the growing curve of CFU-E of infected mice was compared with that of normal mice. Results revealed no significant suppressive effect on CFU-E by P. berghei infection. It is indicated that the maturity and differen-tiation of erythrocytes were the foundation of invasion and parasitism by merozoites.
    ON THE CLASSIFICATION OF SUBGENUS LARROVSIVS NITZULESCU, 1931 and PHLEBOTOMUS WUI STAT.NOV (DIPTERA:PHLEBOTOMIDAE)
    1987, 5(1):  45-48. 
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    The classification of subgenus Larrousius is discussed and the relative importance of their morphological characteristics was compared. The authors stress that the most important taxonomic characteristics is the male aedeagus which varies with the species, while the morphology of the spermatheca segments and the pharyngeal armature are of much importance for female taxonomy. Besides, other morphological characteristics are either useless or of no importance for the classification of subgenus Larrousius. By comparison, P. major Annandale, 1910 and P. major wui Yang et Xiong, 1965 belonging to the subgenus Larrousius, are quite different in their morphological characteristics of pharyngeal armature, spermatheca segments and aedeagus form. Thus it is suggested by the authors that P. major wui Yang et Xiong, 1965 should be raised to species P. wui (Yang et Xiong, 1965) stat. nov. instead of subspecies.
    SCANNING ELECTRON MICROSCOPY ON ANISAKIS TYPE 1 THIRD STAGE LARVA
    1987, 5(1):  49-50. 
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    Surface topography of the Anisakis type I 3rd stage larva (L3), the main pathogen of anisakiasis, was observed by scanning electron microscope under magnifications of 400 to 14000X. The head bears a trapezoid undivided lip mass, with 2 mammillary elevations at the middle of the two lateral sides and a cuticular boring tooth on its ventral side. The mouth leading into the esophagus is situated in the centre of the centrally located cuticular elevation of the lip mass. The excretory pore opens ventrally just behind the boring tooth. The surface of the lip mass shows fine striations, but looks smooth elsewhere. Neither flat papillae nor minute "teeth" or "hairs" described hither to have been observed. The cuticle of the body surface shows shallow and irregular annular grooves and folds and numerous fine longitudinal micro-furrows and cristae. These surface markings appear uniformly from head to tail and to the utmost end of the pagoda like mucrones at the tip of the tail. The ventrally located crescent anus is situated about 90μm from the base of the mucron.
    EXPERIMENTAL INFECTION OF SARCOCYSTIS SUIHOMINIS IN PIGS
    1987, 5(1):  51-52. 
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    Sporocysts of Sarcocystis suihominis collected from feces of infected persons who had ingested raw pork were used to infect swines. On the 56th day post inoculation, cysts were found in the cardiac and skeletal muscles. The cysts are spindle-like, elongated or elliptic. The cyst wall had numerous thick, villi-like projections and the cysts wera divided into many compartments by septa formed from the cyst wall. Each compartment was filled with banana-like bradyzoites, measuring 10.46×4.61 μm. Some of the cysts-had spherical or spheroidal metrocysts.Five rhesus monkeys fed raw pork containing S. suihominis cysts were not infected, while another monkey which had received hydrocortisone injection two days before S. suihominis intubation for 5 days voided sporocysts from feces on the eleventh day post inoculation for a period of 9 days. The sporocysts measured 12.72×10.24 μm.
    A SIMPLE METHOD FOR ISOLATING PLASMODIUM YOELII
    1987, 5(1):  53-54. 
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    20% (W/V) meglumine solution was neutralized to pH7.2 by adding amidotrizoic acid and diluted with saline in a volume ratio of 1:1, 1:1.5, 1:2, 1:3, 1:4 and 1:5 to form different density gradient solutions, then added into a test tube separately.Heparinized infected mouse blood was centrifuged and the blood cell pellet was transferred to the top of density gradient. After centrifugation at 2 500 rpm for 25 minutes, the sample was separated into seven layers, free parasites and the red cells infected with parasites of different stages being in the upper five layers and the uninfected RBC and WBC mainly in the lower two layersThe infected RBC were collected and washed once with saline, then 37 ℃ warmed 0.8% NH4Cl solution was added to lyse RBC membrane. Incubate the suspension at the same temperature for 10 minutes, and then centrifuge down the free parasites at 2 500 rpm. After washing with saline and eliminating the sticky residue in the bottom of the centrifuge tube the purified parasites were harvested, and only one or two white blood cells could be found per 104 parasites with a limited amount of ghost corpuscles.
    PRELIMINARY STUDY ON THE PRODUCTION OF HIGH GAMETOCYTAEMIA OF PLASMODIUM CHABAUDIIN THE RAT
    1987, 5(1):  55-57. 
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    Large numbers of Plasmodium chabaudi gametocytes can be produced in the splene-ctomized Wistar rats 3-4 days after the intravenous inoculation of high parasitaemia mouse blood. On the 8-9th day, the number of gametocytes reduced greatly. The level of gametocytaemia was related to the time of splenectomy and the amount of the infected blood inoculated. The gametocytes in the rat were normal in their ability of exflagellation and their infectivity to anophelines.