Table of Content

31 August 1984, Volume 2 Issue 3

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COMPARISON OF DIFFERENT METHODS OF CIRCUMOVAL PRECIPITIN TESTS (COPT) FOR SCHISTOSOMIASIS JAPONICA

1984, 2(3):  129-132. 

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A research group composing of 7 units to assess the diagnostic value of conventional COPT and its relevant improved methods under the same condition, was organized by the diagnosis Group of the National Schistosomiasis Research Committee, in Changshu City, Jiangsu, in 1983. Indirect hemagglutinatiori(IHA) was used as a control means. Sera of 82 cases of Schistosomiasis japonica having positive stool examination, 51 cases of Schistosomiasis japonica after being cured and having negative stool examination, 51 healthy individuals, 5 cases of clonorchiasis and 4 cases of paragonimiasis, were tested, by double-blindness method. The conventional and improved COPT made on parasitologicajly confirmed cases presented a positive rate of 65.9-85.2 %. The diagnostic effect of enzyme-linked COPT and immunofluorescence COPT appeared to be no higher than that of conventional COPT using lyophilized ova of Schistosoma japonicum. The false positive rate tested for the healthy persons was 2.0-33.3%, that of conventional COPT with lyophilized ova was 2.0-7.8%. The results obtained from the conventional COPT after 72 hours incubation (69.5-84.2%) Were higher than those for 48 hours (65.9-73.2%). In contrast, positive and false positive rates of IHA were 91.5% and 3.9%, respectively, the cross reaction of the patients with paragonimiasis being higher than that of COPT.

EFFECT OF CONTROL MEASURES OF SCHISTOSOMIASIS OBSERVED BY THE ANNUAL INCIDENCE AT A RISK POPULATION IN THE PHILIPPINES

1984, 2(3):  133-135. 

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The annual incidence is a ratio of positives having been converted from negatives during one year. Children at 9 primary schools at Dagami Area, Leyte in the Philip-.pines were followed up from 1974 to 1983 and the incidence rates of schistosomiasis japonica were calculated by the results of fecal examination and COP reactions amngo 1,800 children enrolled in these schools. The survey data were stored, processed and calculated by computer; The observed annual incidence rates were 22.2, 24.2, 26.9, 9.6, 28.4, 8.4, not examined and 6.8% yearly from 1975/76 school year (SY). on. As to control measure, environmental modifications with drainage by. channelling ditches In ;the rice field at the most depressed area of 50 ha in the middle of project area at 5 km radius had been conducted from Dec. 1974 to Mar. 1979. After Feb. 1979, mass chemotherapy with praziguantel has been given to egg positives of children and adults in this area. The cause of abrupt decrease of incidence to 9.6% in SY 1978/79 was thought to be due to less rain fall in the rainy season from Nov. 1977 to Jan. 1978. In the period of environmental modification, the incidence was affected little and was greatly lowered after mass chemotherapy. It was concluded that the small scale environmental modification affected little on reducing danger of infection even to the close area, although it was effective in many places in the world when it was designed and conducted at a large scale.

FIVE YEAR SEROEPIDEMIOLOGICAL SURVEY IN AN AREA WITH SCHISTOSOMIASIS BASICALLY ERADICATED

1984, 2(3):  136-139. 

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In an area with schistosomiasis basically eradicated, circumoval precipitin test (COPT) and enzyme-linked antigen counterimmunoelectrophoresis (ELACIEP) were used as means of seroepidemiological survey to measure the changes of the specific schis-tosome antibody levels for 5 consecutive years in 219-453 subjects having negative ova or miracidia for stool examinations. 60 cases with a history of previous treatment and 53 persons without therapeutic history were also observed by using these two serological methods. The results showed that both COPT and ELACIEP could reveal kinetic changes of specific schistosome antibody levels in seroepidemiological survey, and be used for evaluation of the efficacy of control. Both COPT and ELACIEP were of value in increasing the detecting rates of patients.

PRELIMINARY OBSERVATION ON PRAZIQUANTEL IN THE TREATMENT OF PARAGONIMIASIS SICHUANENSIS

1984, 2(3):  140-143. 

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18 cases of paragonimiasis Sichuanensis were treated by praziquantel from April 1981 to November 1982, and the therapeutic results were evaluated 6 months after treatment. All patients receiving treatment had the history of eating raw crabs or stream water in the endemic area, presented with typical clinical manifestation, and had positive skin test with Paragonimus adult worm antigen. There were 5 cases of cerebro-meningeal type, 1 case of ophthalmic type, 3 cases of pleuro-pulmonary type, and 9 cases of subcutaneous mass type. The dosage schedule of praziquantel was 25 mg/kg three times a day for 2 days for one course. 13 cases received one course, 3 cases of cerebral type and 1 case of ophthalmic type received 2 courses, and one cerebral case received 3 courses of treatment.Since paragonimiasis sichuanensis can not be diagnosed by the finding of eggs in sputum, feces, or body fluids, the only way to evaluate the therapeutic result is by observing the improvement of symptoms and signs. 6 months after treatment, 17 cases had complete absence of symptoms and signs of paragonimiasis; only 1 case had epileptic attacks of markedly reduced frequency and intensity. The side effects of praziquantel were mild and transient. The authors believe that since paragonimiasis sichuanensis is a form of viseral larva migrans, a larger total dosage of praziquantel should be used to effect a cure.

DETECTION OF SCHISTOSOMA JAPONICUM ANTIGENS AND ANTIBODIES BY K-ELISA

1984, 2(3):  144-146. 

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K-ELISA technique was employed to compare the potencies of various schistosome antigens and their corresponding antibodies.Pooled serum was prepared from 3 documented cases of chronic schistosomiasis and served as standard serum. The antigenic potencies of JAMA, SEA, JEU, JEC1, JEC2, and JEC3 were found to be 2.0, 8.0, 4.0, 4.0, 5.0 and 10.0 (AA460/min × 10-2) respectively, JEC3 being apparently the most potent one.The mean serum antibody potencies (AA 460/min× 10-2) of schistosomiasis patients before and after pyquiton treatment were 8.110.3 and 3.010.2 (n = 12) for anti-JAMA; 12.8±0.3 and 9.0 ±2.0 (n = 12) for anti-SEA; 6.2±0.6 and 3.810.4 (n = 30) for anti-JEU; 6.8±0.6 and 4.8±0.4 (n = 30) for anti-JEC2 11.7±0.8 and 2.1±0.4 (n = 30) for anti-JEC3. Those for controls (n = 16) were 0.7±0.7, 1.5±0.5, 2.8±0.6, 1.8±0.2 and 2.0±0.4, respectively.It is suggested that JEC3 might be served as a satisfactory diagnostic antigen and K-ELISA might be a useful laboratory method in the study of schistosome antigens and antibodies.

STUDIES ON THE MICROSOMAL ANTIGENS OF SCHISTOSOMA JAPONICUM Ⅱ. EXTRACTION, FRACTIONATION AND CHEMICAL AND IMMUNOLOGICAL PROPERTIES OF MICROSOMAL ANTIGENS OF ADULT WORM

1984, 2(3):  147-149. 

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Microsome antigens were extracted from homogenate of adult Schistosoma japonicum by differential centrifugation and fractionated by Sepharose CL-2B chromatography. The crude microsome antigen (JAMA-C), the urea soluble fraction (JMC) and the purified microsome antigen (JAMA) were tested with ELISA for their sensitivity and specificity in detecting schistosomal antibody. In kinetic dependent assay (K-ELISA), JAMA showed activity Of 8.8 units with homologous serum. With sera from ascariasis (3), clonorchiasis (3), paragonimiasis (3) and trichinosis (1), all the results were below 1 unit. In microtitre plate ELISA, all the three microsome antigens showed high sensitivity and specificity. The results indicated that the specific activity increased with the process of purification. In comparison with microsome antigen, the cytosol and urea soluble nucleus antigen showed lower activity. When analysed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme linked immunoelectro-transfer blot of SDS-PAGE, JAMA was shown to be a heterogenous material composed of proteins of molecular weight from 14,400 to more than 200,000 daltons and the main antigenic components were beyond 43,000 daltons.

ENDOCRINE CHANGES IN 28 YOUNG PATIENTS WITH SCHISTOSOMIASIS DWARFISM

1984, 2(3):  150-153. 

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Endocrine changes were determined in 28 young patients with schistosomiasis dwar-fism, including 15 girls and 13 boys, with an average age of 16.4 years. Testosterone and estradiol concentrations of male patients were significantly lower than the control values. Testosterone concentrations of female patients were significantly higher than the control values. Estradiol concentrations were lower than normal levels, without statistical significance. LH, F, T, GH concentrations in all patients were within normal limits. LH responses to LRH agonist analogs were abnormal in 2/6 of female patients and 5/7 of male patients. Insufficient synthesis and secretion of somatomedin is probabaly responsible for disturbances of bone and sex ual development. The endocrine function in schistosomiasis dwarfism is different to a certain extent from that in pituitary dwar-fism.

PRELIMINARY STUDY ON MONOCLONAL ANTIBODIES AGAINST SCHISTOSOMA JAPONICVM

1984, 2(3):  154-157. 

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The fusion between myeloma cells from BALB/c mice and spleen cells of mice infected with Schistosoma japonicum has been shown to be a successful method for deriving hybridoma antibodies. Indirect immunofluorescence assay on sections of adult worm or schistosomula has successfully been used as an initial method for screening specific antibodies. Three different patterns of immunofluorescence reaction were found in the present study. Two hybrid clones have been obtained and their immunological specificities were determined. One showed positive reaction to certain body cells of the adult worm and schistosomula. Its immunoglobulin subclass was determined to be IgG1. The other showed positive reaction to the digestive tract of the adult worm but negative to schistosomula. Its immunoglobulin subclass was IgG3.

CIRCUMOVAL PRECIPITIN TEST IN PATIENTS INCONTROLLED AND UNCONTROLLED REGIONS OF SCHISTOSOMIASIS JAPONICA

1984, 2(3):  158-160. 

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COP and fecal egg count were carried out on 324 proven cases in three endemic regions of schistosomiasis japonica of different status of snail infestation: 1. uncontrolled region, where Oncomelania is still present; 2. region of minimal transmission, where the snail has not been found for only one year; 3. controlled region, where snail has been eradicated for more than six years. The results revealed high sensitivity of COP for schistosomiasis japonica even in controlled region (97.8% positive among 46 patients) as well as in uncontrolled one (99.0% among 104) and in region of minimal transmission (96.2% among 174). But the intensity of the positive reactions was very different in the three regions, with 84.8% of the sera showing slightly reactive(+) in the controlled region, and 70% of the sera showing moderate or strong reactions (++ and +++). The intensity of infection of the patients was directly correlated with the percentage of COP egg reactivity in the three regions. It is interesting to note that the antibody level (represented by % COP egg reactivity) may decline slowly in the chronic patient even without treatment. Therefore in the controlled region, the COP egg reactivity of more than 1% might have diagnostic value for schistosomiasis.

PRELIMINARY OBSERVATION ON THE ACTIVITY OF T LYMPHOCYTE ACID α-NAPHTHYL ACETATE ESTERASE IN BLOOD OF PATIENTS WITH SCHISTOSOMISIS JAPONICA

1984, 2(3):  161-163. 

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31 blood samples from proven patients with chronic schistosomiasis, 35 from the late stage patients, and 37 from healthy persons were tested by both T lymphocyte esterase label test and lymphocyte transfoamion test (LTT). Both ANAE-label test positivity rate and lymphocyte transformation rate were significantly lower in the schistosomiasis patients than in the healthy persons. The results measured by both methods were closely related.It is preliminarily considered that the determination of blood ANAE activity may be taken as a reference index for the evaluation of cell immunity function of the patients with schistosomiasis.

EXPERIMENTAL THERAPY OF HEPATIC FIBROSIS IN SCHISTOSOMIASIS JAPONICA II. EFFECT OF COLCHICINE ON HEPATIC FIBROSIS

1984, 2(3):  164-166. 

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28 rabbits, each infected percutaneously with 80 cercariae of Schistosoma japonicum developed pipe-stem fibrosis resembling late human schistosomiasis 4 months later. After a single oral dose of pyquiton 100mg/kg, colchicine was then given to 9 rabbits orally to treat schistosomal hepatic fibrosis in a dosage of 1 mg daily for 3 months. The remaining 19 rabbits were used in different groups of controls, namely, infected control with pyquiton 13, infected control without pyquiton 6, uninfected control 6. Liver biopsies were made before and after colchicine treatment for collagen quantitative determination and pathological observation. The results showed that collagen content was reduced from 43.07±4.25 mg/g wet weight to 15.79±2.66 mg/g wet weight, the thick fibrous bands around intrahepatic portal zone were markedly reduced, and degenerated liver cells mostly recovered. In comparison with amygdalin, hydrocortisone and other Chinese traditional medicines in our previous work, colchicine appears to be a more effective antifibrotic drug.

EFFECT OF ARTEMETHER ON TEGUMENTAL SURFACE ANTIGEN OF SCHISTOSOMA JAPONICUM

1984, 2(3):  167-169. 

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This paper deals with the alteration of tegumental surface of Schistosoma japonicum in the mice after oral administration of artemether suspension. Some of the female and male worms showed positive reaction in the indirect fluorescence antibody assay 1 and 5 days respectively after a single dose of 300mg/kg. Different degrees of intensity of fluorescent reaction were demonstrated in all mature female and about one-half of male worms on day 5 and day 9 respectively, and a part of bisexual worms still showed some positive reaction on day 12. The time of exposure of tegumental surface antigen, the number of worms so affected and the intensity of fluorescent reaction of worms were related to the dosage of drug administered. On day 5 after medication, none of the immature female worms showed positive reaction.

CULTIVATION OF GIARDIA LAMBLIA AND CERTAIN OBSERVATIONS ON ITS BINARY FISSION PROCESS

1984, 2(3):  170-172. 

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Certain characteristics concerning the growth and multiplication of Giardia lamblia were observed in the course of monoxenic and axenic cultivation of the parasite. The organism used for monoxenic culture originated from a local giardiasis patient. Cysts from patient fecal specimen were isolated by sucrose gradient centrifugation and then used to infect rabbits. Trophozoites recovered from intestinal mucosa of the infected animal were cultivated at 37℃ together with a small amount of Saccharomyces cerevisiae. They grew luxuriantly and formed a monolayer upon the wall of the culture tube at about 24 hours. Thereafter, yeast cells gradually overgrew the parasite whose number becomes less and less, until very few remains at the fifth day.Strain of G. lamblia and TPS-1 medium used in the axenic culture was supplied by Dr. Visvesvara (CDC, Atlanta). Certain characteristics of the multiplication of the parasite, particularly those concerning the binary fission process, were observed under scanning electron microscope and optic microscope during the course of cultivation which lasted 5 months. The morphological changes at various stages of binary fission of the organism were described.

IMMUNOFLUOHESCENCE CIRCUMOVAL PRECIPITIN TEST IN THE DIAGNOSIS OF SCHISTOSOMIASIS JAPONICA

1984, 2(3):  173-175. 

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A comparative study on the diagnostic value of immunofluorescence technic(IFCOP) and the conventional COP test using heat-treated and ultrasonicated dry eggs of Schisto soma japonicum as antigens for the diagnosis of schistosomiasis japonica was carried out in areas of different endemicity of schistosomiasis japonica. Results showed no significant difference in sensitivity between two methods in the area of high prevalence. However, in the area of low prevalence IFCOP technic was significantly more sensitive than COP test. Results of tests on sera from 63 patients of schistosomiasis showed that IFCOP and COP were positive in 88.9% and 66.7% respectively. The results indicate that the sensitivity of the COP test is much enhanced by the fluorescent antibody technique in that the small circumoval precipitates can be detected more easily.

IMMUNOGENICITY OF SECRETION MATERIALS OF ACETABULAR GLANDS OF SCHISTOSOMA MANSONI CERCARIAE

1984, 2(3):  176-178. 

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As cercarial secretion material (CSM) from the acetabular glands is the first subs-tance to contact the host during infection, the question is raised whether the CSM is immunogenic. We have compared the immunizing abilities of newly-transformed schistosomula-like organisms with intact CSM and with schistosomula in which the CSM has been depleted by prolonged cultivation. A Puerto Rican strain of Schistosoma mansoni was used. Female CF1 mice were immunized by the injection of schistosomula which had been transformed from X-irradiated cercariae at 36kR. CSM-intact schistosomula-like organisms were used for immunization immediately after their transformation by the syringe technique. CSM-depleted schistosomula were prepared by incubating them in Earlc's saline with 0.65% lactabumin hydrolysate in a CO2 incubator at 37℃ for 14 hrs. Purpurin and PAS staining showed that both the pre- and post-ace tabular glands were depleted of CSM in the cultured, but not in the uncultured schistosomula. For each immunization, 500 schistosomula-like organisms or schistosomula were injected intramuscularly into the right hind limb. Mice were immunized 3 times at 4 week intervals, then challenge-infected with 100 cercariae 30 days after the last immunization. Age-matched unimmunized control mice were similarly exposed to 100 challenge cercariae. Worms were recovered by perfusion 41-44 days after challenge.Fifty-three mice were divided into three groups. GroupⅠ consisted of 18 unimmunized controls; Group Ⅱ, 20 mice immunized with CSM-intact schistosomula-like organisms; and Group Ⅲ, 15 mice immunized with CSM-depleted schistosomula. Mean worm recoveries from the challenge infections were 50.5 (±13.8), 21.9 (±11.0), and 28.9 (±8.7) for Groups Ⅰ,Ⅱand Ⅲ, respectively. Analysis of variance on arcsin transformed data indicated significant differences among the three group means (F= 32.82, P0.001). Duncan's test showed significant differences among all three means at the 5% level. Worm reductions were 56.6 and 42.8% in Groups Ⅱ and Ⅲ, respectively. The results of the present study show that CSM is a protective immunogenic substance in this case, but it should be considered only as an auxiliary immunogen, because CSM-depleted schistosomula were also protective, though to a less extent.

PRELIMINARY APPLICATION OF ELISA IN LIVER SECTIONS OF INFECTED MICE FOR THE DIAGNOSIS OF SCHISTOSOMIASIS JAPONICA

1984, 2(3):  182-184. 

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This article presents the application of ELISA to liver sections of infected mice containing schistosomal egg granuloma as antigen to detect the specific antibody in sera of schistosomiasis patients. 56 sera from patients with acute (35) and chronic(21) schistosomiasis japonica were all positive, while 3 of 6 patients with advanced schistosomiasis showed positive. No false positive reaction was found in 50 controls. The technique appears quite sensitive and specific in the diagnosis of schistosomiasis japonica.

ASSAY OF SUPEROXIDE DISMUTASE ACTIVITY IN THE PERIPHERAL BLOOD CELLS OF PATIENTS WITH SCHSTOSOMIASIS

1984, 2(3):  185-187. 

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SOD activity levels in the peripheral blood cells of 19 cases of early schistosomia- sis, 43 cases of late schistosomiasis, 9 cases of late schistosomiasis who have been essentially cored and 30 cases of healthy persons wfere determined.The results showed that SOD activity is 98 3U in patients with early schisto-miasis, 486U in patients of late schistosomiasis, 617U in patients of late schistosomiasis who have been essentially cured, and 1,411U in healthy persons, respectively.There exists a significant difference between the SOD activity in normal healthy persons and in patients of late schistosomiasis (P0.001).

OBSERVATION ON FECAL EGG COUNT IN 10 SCHISTOSOMIASIS JAPONICA CASES FOR 16 MONTHS

1984, 2(3):  188-189. 

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An observation on fecal egg count with thick-sediment smear method was carried out in 10 schistosomiasis japonica cases in Lumu Commune Wuxian County neatfSuzhou City. 3 counts monthly for 16 successive months were done in each patient. The data were)transferred to logarithmic values and treated with analysis of variance method. Result showed there was no significant difference between the geometric mean values of the fecal egg counts detected in different months, but significant difference was found; between the geometric mean values of the fecal egg counts in individual patients. It is presumed that the amount of schistosome eggs expelled in feces was more or less constant in a certain period of time. Fecal egg count seemed, therefore, practical in the study of epidemiology and surveillance of control effect of schistosomiasis japonica, and may be used freely in any month.

PATHOLOGICAL CHANGES AND CLINICAL TYPES IN ADVANCED SCHISTOSOMIASIS

1984, 2(3):  190-192. 

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This paper reports on the hepatic pathological changes in 106 clinically diagnosed advanced schistosomiasis japonica cases from 1977-1981 in our hospital. The specimens were obtained from liver biopsy during surgical operation. After pathological examination, 67 cases (63.2%) were classified as cirrhosis of simple schistosomiasis and 39 cases (36.8%) as cirrhosis of scfeistosomiasis complicated with viral hepatitis. The authors described in detail about the pathological morphology of the above two types of cirrhosis and suggested to have liver biopsy during splenectomy of the patients in order to make accurate diagnosis and give corresponding effective therapy.