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PubMed1986

Immunodiagnosis of Entamoeba histolytica and Ascaris lumbricoides infections in Caucasian and aboriginal Australians.

The immunodiagnostic efficiency of an indirect immunofluorescence test (IFAT) and in vitro lymphocyte proliferative responsiveness (cell mediated immunity test, CMIT) used to measure the immunological responses of individuals with known natural Entamoeba histolytica and Ascaris lumbricoides infections, was studied under survey conditions. E. histolytica was common among Aborigines from Cherbourg, Kowanyama and Central Australia, but it was not found in Brisbane Caucasians. The protozoan was selected for the study because it was prevalent and purified antigen was commercially available. Immunodiagnosis for A. lumbricoides was made using an antigen prepared by affinity chromatography. Diagnosis based on frequency distribution of immunological data gave valid assessment of the number of infected individuals in each population studied.

Adolescent↗
PubMed1986

A dot immunobinding assay (DIA) on nitrocellulose membrane for the serological detection of antibodies to Entamoeba histolytica.

A rapid, cheap, simple and specific serological test of adequate sensitivity for detecting IgG antibodies against Entamoeba histolytica antigen is described. Axenically cultured amoebic antigen was used to precoat the nitrocellulose membrane. The strips were incubated with test samples and later with horseradish peroxidase (HRPO) labelled protein-A conjugate. A dark blue spot was obtained by treatment with peroxidase substrate, 4-chloro-1-naphthol, in positive samples. Serum samples from 32 healthy controls, 45 patients with acute amoebic liver abscess and 10 asymptomatic E. histolytica cyst passers were tested. This test was positive in 93% of cases of amoebic liver abscess, 3% of healthy controls and none of the cyst passers; its sensitivity (97%) and specificity (93%) were as good as that of the ELISA test and, because it is simple, quick and cheap it is recommended as a serological test of choice for the diagnosis of invasive amoebiasis.

Antigens, Protozoan↗
PubMed1987

Low susceptibility of trophozoites of virulent Entamoeba histolytica to cellular and antibody-dependent cellular cytotoxicity by guinea-pig effector cells.

Cytotoxic potentialities of lymphocytes and macrophages were determined against trophozoites of virulent and avirulent sublines of Entamoeba histolytica in vitro. Guinea-pigs were immunized with antigens of both sublines to obtain stimulated effector cells and antiamoebic antibodies. Trophozoites of an avirulent/attenuated subline of E. histolytica (NIH200) were significantly more susceptible to killing by lymphocytes and macrophages through cellular and antibody-dependent cellular cytotoxic mechanisms. The resistance of virulent trophozoites to killing was attributed to their higher phagocytic ability and virulence characteristics. Electron microscopic studies of the interactions showed that contact between the effector cells and the trophozoites was essential for the cytolytic event which resulted in disintegration of the trophozoites. Antigens of trophozoites of the virulence subline of E. histolytica (NIH200 V) were effective in inducing cytotoxicity against both virulent and avirulent trophozoites: however, the reverse was not true.

Animals↗
PubMed1987

Entamoeba histolytica in inbred mice: evidence of liver involvement.

Following observations that certain mouse strains appeared to be susceptible to liver invasion by Entamoeba histolytica, mouse strains CBA/HN, CBA/HN, nu/nu, CBA/HN nu/+, CBA/HN Dh/+, CBA/HN +/+, C3H/HeJ Lps/Lps and AB/H, were infected intra-intestinally with strain SAW 408 (zymodeme II) of the amoeba. A number of mice from each group died during the first 14 days with acute lesions. The remainder were killed at 4 and 6 months after infection. CBA/HN, CBA/HN Dh/+, and CBA/HN +/+ mice all had amoeba-infected livers, although only 2 mice had extensive, typical liver lesions.

Animals↗
PubMed1987

The effect of orally administered Entamoeba histolytica on Wistar and athymic (rnu rnu) rats observed during a 12-month period.

Groups of LacP (Wistar) and rnu rnu athymic rats were dosed orally with cultured trophozoites of 2 strains of Entamoeba histolytica, zymodeme II, and observed for up to 12 months. One group of Wistar rats remained infected but with no apparent disease for 6 months; faecal samples from another became negative after one week. All the athymic rats were still infected at 12 months and amoebae could be cultured from their stools. The effect of even long-term infection was usually only mild erosion of the caecal wall and only 2 rats developed typical chronic lesions and ulcers. Amoebae were isolated in culture from the apparently unaffected livers of 2 rats 12 months after infection.

Animals↗
PubMed1987

An enzyme-linked immunosorbent assay for the detection of Entamoeba histolytica antigens in faecal material.

This paper describes a method for the detection of Entamoeba histolytica antigens in stool samples using a multi-layer ELISA. The method is sensitive and specific, showing no interference with other intestinal parasites, e.g. E. coli, E. hartmanni, Endolimax nana, Iodamoeba buetschlii, Hymenolepis nana, Giardia lamblia, Trichomonas and Ascaris. The method provides a rapid and simple screening assay for E. histolytica infections and should assist in diagnosis and epidemiological studies of the disease.

Animals↗
PubMed1988

Biological evidence of genetic exchange in Entamoeba histolytica.

The demonstration of a new zymodeme of Entamoeba histolytica produced in culture from cloned isolates suggested possible genetic exchange in this parasite. We have attempted to substantiate that finding by using rats as biological hosts. Clones were made from 3 separate isolates of E. histolytica, each established in culture from liver pus or faeces. After enzyme characterization these clones, of zymodemes II, XIV and XIX, were paired in each of the 3 possible combinations and the mixtures injected into the caecum of rats. Clones of new or hybrid zymodemes were produced as well as the original parents, with one exception, the mixture of XIV and XIX, from which only one of the parents was recovered. The hybrids produced included zymodeme XX, observed previously, and zymodeme XI, a naturally occurring zymodeme that has in the past been recovered only from subjects with invasive amoebiasis.

Animals↗
PubMed1988

The prevalence of Entamoeba histolytica in lactating women and in their infants in Bangladesh.

Entamoeba histolytica was studied in 33 lactating women and their infants in a periurban village in Bangladesh. Infant-mother pairs were followed for a period of 10-15 months: 67% of mothers excreted E. histolytica during the observation period, the majority for 3 months or more. Only one mother was symptomatic, with a mild, non-dysenteric diarrhoea. 58% of mothers were seropositive, several of them continuously and with a high titre, indicating past invasive infection. 67% had detectable antibodies in breast milk and 36% in saliva. Despite the high prevalence of E. histolytica in these mothers, infants were mostly uninfected: E. histolytica cysts in small numbers were found in only 2 of 1200 samples from infants aged 6 and 10 months. Several of the children were infected with Giardia lamblia. Although lower exposure to E. histolytica than to Giardia may account for the difference in infant infection rates with these parasites, defence mechanisms possibly exist which protect against E. histolytica but are ineffective against Giardia.

Adolescent↗
PubMedDate not supplied

High susceptibility of five axenic Entamoeba histolytica strains to gossypol.

The antiamoebic potency of gossypol was tested against 5 axenic Entamoeba histolytica strains, in logarithmic phase growth in PEHPS medium. All of the strains were moderately susceptible to this polyphenolic drug. The 50% inhibitory concentration (IC50) was of the same order of magnitude in all strains: 0.015 microM (for strain HM-1) to 0.067 microM (for HM-38). The difference between the IC50 of HM-1 and the remaining 4 strains (HM-38, HK-9, HM-2 and HM-3) was significant, although it was greater between HM-1 and HM-38 (P less than 0.005) than between HM-1 and the other 3 strains (P less than 0.05), for which the IC50 was 0.03 microM, 0.049 microM and 0.38 microM respectively. Gossypol is more toxic in vitro for amoebae than other drugs widely used clinically, its pharmacological effects and safe dosage in humans are well known because of its antifertilizing effect, and it is accumulated mainly in the liver and colon. Accordingly, this compound could be a good antiamoebic agent if it is as potent in vivo as it is in vitro.

Animals↗
PubMedDate not supplied

Experimental production of new zymodemes of Entamoeba histolytica supports the hypothesis of genetic exchange.

By mixing in culture zymodemes of Entamoeba histolytica, clones showing a different pattern from the 2 parents have been obtained on 3 occasions. Two new zymodemes (XI alpha- and XXI) have thus been produced, which belong to the third generation of a family of E. histolytica zymodemes apparently obtained by genetic exchange. A hypothesis for the molecular basis of those exchanges is discussed. We report also the observation of probable mutation.

Animals↗
PubMedDate not supplied

Virulence of pathogenic and non-pathogenic zymodemes of Entamoeba histolytica (Indian strains) in guinea-pigs.

Guinea-pigs were inoculated with Entamoeba histolytica strains isolated from cases of amoebic liver abscess, amoebic dysentery and asymptomatic cyst passers and the strains were classified in zymodemes by isoenzyme electrophoresis. It was observed that certain non-pathogenic zymodemes were potentially pathogenic for the guinea-pigs. The relationship of zymodemes and virulence in laboratory animals is discussed.

Amebiasis↗
PubMedDate not supplied

In vitro activity of nalidixic acid and its iron (III) complex on Entamoeba histolytica.

The in vitro activity of the antibacterial agent nalidixic acid (HNal) and its iron (III) complex (FeNal) against Entamoeba histolytica HM1 strain trophozoites in axenic or monoxenic (associated with Clostridium symbiosum) cultures was investigated. Using a dilution test with TYI-S-33 medium, this protozoan was found to be susceptible to both drugs, but FeNal showed amoebicidal activity only at concentrations higher than those used with HNal.

Animals↗
PubMedDate not supplied

Entamoeba histolytica infections in flight personnel of an international airline.

A cohort of 99 subjects, members of the flight staff of Lufthansa German airlines, all infected with Entamoeba histolytica, were followed over a period of 2.5 years to monitor their clinical and parasitological status. All but 4 of the subjects were infected with non-pathogenic zymodemes of E. histolytica and had negative amoebic serology, and most were clinically well. One male heterosexual carrying a pathogenic zymodeme was found, despite massive treatment regimes, to carry the infection persistently for 12 months. Most non-pathogenic infections were left untreated.

Aircraft↗
PubMedDate not supplied

Zymodeme conversion of isolates of Entamoeba histolytica.

A non-cloned and a cloned isolate of non-pathogenic (zymodeme I) Entamoeba histolytica derived from homosexual males were incubated in Diamonds' TP-S-1 medium in the presence of Crithidia for 52 d. A single addition of Escherichia coli irradiated at 1000 Gy rendered the culture axenic. Analysis of zymodeme patterns established that both isolates had transformed to zymodeme II. The re-establishment of cultures in the presence of microflora from the original cultures resulted in a return to zymodeme I, whereas incubation with microflora from an amoebic isolate exhibiting a zymodeme II pattern maintained the pathogenic pattern.

Animals↗
PubMedDate not supplied

A modification of Diamond's medium for the axenic culture of Entamoeba histolytica.

The use of casein hydrolysate in Diamond's axenic culture medium TPS-1 in replacement of trypticase allowed good growth of the trophozoites of Entamoeba histolytica. This modified medium also supported growth of trophozoites preserved for 16 months in liquid nitrogen. Considerable labour and cost of serum can be saved by using 5% instead of 10% bovine serum in combination with this modified medium.

Animals↗
PubMedDate not supplied

High cell surface hydrophobicity of virulent Entamoeba histolytica isolates.

Thirty isolates of Entamoeba histolytica were examined for their cell surface hydrophobicity. It was observed that increased hydrophobicity of the trophozoites was associated with lesion-forming ability in hamster liver, a higher rate of erythrophagocytosis and greater resistance to complement. The relationship of cell surface hydrophobicity to other virulence markers in E. histolytica is discussed.

Animals↗
PubMedDate not supplied

Enzyme-linked immunosorbent assay based on monoclonal and polyclonal antibodies for the detection of Entamoeba histolytica antigens in faecal specimens.

A murine monoclonal antibody (MAb) (Eh208C2-2) raised against crude lysate of the pathogenic HM-1:IMSS strain of Entamoeba histolytica was used in an enzyme-linked immunosorbent assay for the detection of E. histolytica antigens in faecal specimens. The detection limit of the assay was 110 and 280 amoebae/ml of the HM-1:IMSS and HK-9 strains in phosphate-buffered saline, respectively. The assay was applied to single stool samples from 3 groups of individuals comprising 40 patients whose stools were positive for E. histolytica trophozoites and/or cysts (group I), 48 patients whose stools were negative for E. histolytica but positive for other parasites (group II), and 36 parasitologically-negative healthy controls (group III). Positivity rates of 77.5%, 2.1% and 2.7% were found in samples from groups I, II and III respectively. Specificity, positive and negative predictive values, and efficiency of the assay were 97.6%, 93.9%, 90.1% and 91.1% respectively. When group I samples were further divided into a trophozoite-positive subgroup IA (13 samples) and a cyst-positive subgroup IB (27 samples), the positive rates were 100% and 66.7%, respectively (P < 0.025).

Animals↗
PubMedDate not supplied

Entamoeba histolytica: erythrophagocytosis, collagenolysis, and liver abscess production as virulence markers.

High rates of erythrophagocytosis and collagenolysis in vitro have been regarded as indicative of virulence in vivo of Entamoeba histolytica trophozoites. In the present study, the erythrophagocytic index and the collagenolytic activity of 3 axenic lines of E. histolytica, strain HM1:IMSS, were measured. The 3 lines shared the same pathogenic zymodeme but showed clear-cut differences in the extent of liver damage induced in hamsters. A direct correlation between collagenolysis in vitro and the size of liver abscesses produced by each line of E. histolytica trophozoites was found. In contrast, the line with the highest erythrophagocytic index produced small amoebic abscesses in hamsters, whereas the line with a relatively low erythrophagocytic index produced the largest liver lesions. It is concluded that the extent of collagenolytic activity is a better marker of virulence of E. histolytica cultured under axenic conditions than is erythrophagocytosis.

Animals↗