Clustering of Ascaris lumbricoides and Trichuris trichiura infections within households

A survey of 428 households in a shanty town in Coatzacoalcos, Mexico, revealed high prevalences of Ascaris lumbricoides and Trichuris trichiura. The data were analysed separately for A. lumbricoides and T. trichiura in order to investigate the spatial distribution of heavy infections through the town. Within each age class, those individuals with egg counts in the upper 20% of the range for that age were classified as "heavily infected". When the data were stratified by household size, it was found that the distribution of "heavily-infected" individuals was not random in the community. "Heavily-infected" individuals were found together in households; fewer household units had a single heavily-infected individual than would be expected by chance. Such a pattern could result either from genetic similarities among family members influencing their ability to mount an effective immunological response to infection, or focal transmission in the vicinity of the home, or both. This result may have important implications for the development of community control programmes.     

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.     

Risk factors for infection by the Entamoeba histolytica/E. dispar complex: an epidemiological study conducted in outpatient clinics in the city of Manaus, Amazon Region, Brazil

An epidemiological study was conducted on a population attending outpatient clinics in Manaus, Amazon, Brazil to determine the prevalences of infection by the Entamoeba histolytica/E. dispar complex and by E. histolytica alone, as well as to identify the risk factors involved in transmission. The study was conducted in two phases: survey and case-control. Face-to-face interviews were carried out and faecal samples collected from 1578 individuals. Faeces were examined by optical microscopy and tested for the pathogenic E. histolytica specific antigen. Positivity to E. histolytica/E. dispar was 21.5% (340 cases). Cases were compared with 340 control samples, negative for the E. histolytica/E. dispar complex based on examination by optical microscopy. The analysis was conducted by logistic regression. The risk factors identified were: place of residence, age, ingestion of raw vegetables, quality of water consumed, number of rooms and bedrooms per house, and having other protozoan infections. Specific antigen detection tests identified 22 participants infected by E. histolytica (6.8%) among those positive for E. histolytica/E. dispar. There was a higher proportion of males among participants infected by E. histolytica than among those with E. dispar infections. The study population was asymptomatic or presented non-specific symptoms that could be attributed to amoebiasis.     

Prevalence and risk factors for protozoan and nematode infections among children in an Ecuadorian highland community

In order to create a parasite prevention program for Santa Ana, Ecuador, we worked with local physicians to identify the prevalence and risk factors for intestinal parasitic infection among children. Two hundred households were randomly selected and asked to submit three stool samples from a selected child. Information from direct observation and questionnaires was used to determine environmental and behavioral risk factors for parasitic infection. Prevalence of any parasitic infection was 65.6% of the 189 children who submitted fecal samples. Parasitic infections included Entamoeba histolytica/E. dispar (46.6%), Giardia lamblia (18.5%) and Ascaris lumbricoides (20.1%). In a multivariate logistic model, risk factors (P<0.05) for E. histolytica/E. dispar included >1 month since antiparasitic treatment, not adding chlorine to drinking water, children not washing their hands before eating, and having a fewer number of electrical appliances in the home. The risk factor identified for G. lamblia was children not washing their hands before eating. Risk factors for A. lumbricoides included >6 months since antiparasitic treatment and having a fewer number of electrical appliances in the home. This study may have implications for other communities throughout the developing world.     

Prospective evaluation of lactose malabsorption by lactose hydrogen breath test in individuals infected with Entamoeba histolytica and passing cysts

The aim of the present prospective study was to detect lactose malabsorption in subjects in northern India infected with Entamoeba histolytica and passing cysts. The study group included forty-one patients with E. histolytica cysts in at least one of three consecutive faecal samples. Lactose malabsorption was detected by a lactose H2 breath test. The results were compared with those of forty controls subjects. Thirty-two of forty-one (78.0 %) subjects passing E. histolytica cysts had lactose malabsorption compared with seventeen of forty (42.5 %) control subjects (P<0.01). In conclusion, the present study shows that lactose malabsorption is significantly more common in individuals infected with E. histolytica and passing cysts compared with control subjects.     

Use of enzyme linked immunosorbent assay in intestinal and extra-intestinal amoebiasis (amoebic liver abscess)

The sera of patients with amoebic liver abscess, intestinal amoebiasis and non-specific hepatomegaly were examined for the presence of antibodies to Entamoeba histolytica antigen by enzyme linked immunosorbent assay (ELISA). All the amoebic liver abscess sera were strongly positive. The test was also positive in all the proved cases of intestinal amoebiasis but the ELISA values were relatively slightly lower. Nine of 10 cases of non-specific hepatomegaly with stools negative for E. histolytica gave positive results with ELISA. The IHA titre in all but one of the non-specific hepatomegaly cases was within normal value.     

Enzyme linked protein-A: an ELISA for detection of amoebic antibody

Enzyme linked protein-A was used to develop an enzyme linked immunosorbent assay (ELISA) system for the detection of circulating antibodies to amoebic antigen. The specificity of protein-A to bind IgG only through Fc receptors, makes the test more specific for the detection of IgG antibodies to amoebic antigen. The ELISA system was used to detect amoebic antibody in control subjects (56), patients with amoebic liver abscess (79) and Entamoeba histolytica cyst-passers (10) and the results compared with those of indirect haemagglutination assay (IHA). The ELISA was more sensitive and detected 74.7% of cases with amoebic antibody in amoebic liver abscess compared with 66.7% detected by IHA. The test was more specific, sensitive and easy to perform and is recommended as a test of choice for the serological diagnosis of amoebic liver abscess.     

Intestinal parasitism and Vibrio cholerae infection among diarrhoeal patients in Kolkata, India

In this study, we have analysed the epidemiological significance of the concurrent infections caused by Vibrio cholerae and intestinal parasites among different age groups of hospitalized diarrhoeal patients in Kolkata. A total of 3556 stool samples collected during 1996-2004 were screened for vibrios and parasites. The seasonality of V. cholerae and parasitic infections were studied in detail. The detection rates for Ascaris lumbricoides and Giardia lamblia infection were more than for the hookworm, Trichuris trichiura and Entamoeba histolytica. V. cholerae O1 was identified as the predominant serogroup among diarrhoeal patients. The highest rates for V. cholerae infection were in the 2-5 years age group and combined infection of V. cholerae and parasites was recorded among children aged between 2 and 10 years.     

Antibody and cytokine responses in Dracunculus medinensis patients at distinct states of infection

Dracunculiasis is a promising candidate for eradication, but transmission of Dracunculus medinensis and recrudescence of the disease have been observed repeatedly. In the present investigation, the D. medinensis-specific cellular cytokine response profiles and the parasite-specific antibody subclass reactivity were evaluated in dracunculiasis patients at distinct states of infection. Analysis of the cellular cytokine response in dracunculiasis patients disclosed a D. medinensis antigen-specific depression of IFN-gamma production with patent D. medinensis infection, while the T helper type 2 cytokine IL-10 was similar in patent, post-patent and control individuals, and IL-5 production was always the highest in controls. In parallel, diminished IFN-gamma and IL-12 responses to antigens from Ascaris lumbricoides, Entamoeba histolytica and mycobacteria were observed in patent and post-patent dracunculiasis cases. The parasite-specific IgG(1) and IgG(4) subclass reactivity profiles corresponded with the D. medinensis infection state, and a clear distinction between patent and post-patent patients and controls was found. Overall a depressed cytokine release was observed with patent D. medinensis, which extended beyond the parasite-specific immune responsiveness. The detection of D. medinensis-specific IgG(1) and IgG(4) isotypes may help to distinguish newly exposed, patent and post-patent D. medinensis infections.     

Species-specific field testing of Entamoeba spp. in an area of high endemicity

Entamoeba histolytica has been separated in recent years into 2 morphologically identical species: the apathogenic E. dispar and the pathogenic E. histolytica, only the latter being pathogenic. Although various laboratory techniques allow discrimination between the 2 species there is a lack of field data about the suitability of available diagnostic tests for use in epidemiological studies and few epidemiological studies using species-specific diagnosis have been performed at community level in endemic areas, especially in sub-Saharan Africa. We conducted a repeated cross-sectional study of 967 schoolchildren in central C?te d'Ivoire to compare and evaluate light microscopy, 2 different antigen detection assays, and one polymerase chain reaction (PCR) assay. Microscopy and a non-specific antigen capture Entamoeba enzyme-linked immunosorbent assay (ELISA) were used for the primary screening of all children (time t0). The prevalence of the E. histolytica/E. dispar species complex at t0 was 18.8% by single microscopical examination and 31.4% using the non-specific ELISA. Approximately 2 months after the initial screening, fresh stool specimens were collected on 2 consecutive days (t1 and t2) from (i) all the children who were positive by microscopy at t0 (n = 182) and (ii) 155 randomly selected children who were negative at the primary screening. These samples were tested with a second antigen detection ELISA specific for E. histolytica (n = 238) and with a species-specific PCR assay (n = 193). The second and third examinations (t1 and t2) revealed an additional 43 infections with the species complex E. histolytica/E. dispar, so that the cumulative microscopical prevalence for t1 and t2 was 27.7%. The overall prevalence of E. histolytica by species-specific ELISA antigen detection was low (0.83%), while the prevalence of E. dispar was 15%. When analysing only microscopically positive samples by PCR (n = 129), the ratio E. histolytica: E. dispar was very low (1:46), suggesting that the vast majority of Entamoeba infections in this area were apathogenic. Both species-specific tests performed well but the ELISA was easier to use for large-scale field screening.     

Immune responses to an inactive vaccine against American cutaneous leishmaniasis together with granulocyte-macrophage colony-stimulating factor

The immunological response in healthy subjects to a crude leishmania antigen vaccine (Leishvacin) plus rhGM-CSF without prior Montenegro (DTH) skin testing was evaluated. Fifty-six healthy volunteers received vaccine plus either placebo or rhGM-CSF at day 0, followed by either a vaccine booster or placebo at day 21. IFN-gamma and IL-5 levels were significantly enhanced by day 21. The adjuvant group had a higher percentage of individuals with a significant response to vaccination than the corresponding placebo group. Eighty-six percent of all volunteers were DTH-positive by day 42. Leishvacin is capable of sensitizing lymphocytes from individuals not previously exposed to leishmania antigen. Use of rhGM-CSF enhanced the immune response, indicating that it may improve immunological response to the vaccine.     

The risk of enteric infections associated with wastewater reuse: the effect of season and degree of storage of wastewater

The effect of season and wastewater storage on the risk of Ascaris lumbricoides infection and diarrhoeal disease associated with wastewater reuse was studied in Mexico in 1991. Data were collected from 10,489 individuals during a dry-season survey. Exposure was to untreated wastewater, or effluent from 1 reservoir (< or = 1 nematode egg/L), or no wastewater irrigation (control group). The results were compared with a previous rainy-season survey which included effluent from 2 reservoirs in series. Direct exposure to untreated wastewater was associated with an excess risk of A. lumbricoides infection in children aged < 5 years (OR = 18.0) and persons aged > 5 years (OR = 13.5) and an increased risk of diarrhoea, particularly to children aged < 5 years (OR = 1.75); effects were stronger in the dry than in the rainy season. There was also an excess risk associated with the 1-reservoir group for A. lumbricoides infection (OR = 21.2 and 9.4) and for diarrhoeal disease (OR = 1.1 and 1.5) but little excess associated with the 2-reservoirs group. Therefore, wastewater retention in 1 reservoir (quality 10(5) faecal coliforms/100 mL, < or = 1 egg/L) does not significantly reduce risks of Ascaris infection and diarrhoeal disease whereas retention in 2 reservoirs in series (quality 10(3) faecal coliforms/100 mL, no detectable eggs/L) does.     

Health impact evaluation of wastewater use in Mexico.

Wastewater from Mexico city is used to irrigate over 85 000 hectares, mainly of fodder and cereal crops in the Mezquital Valley. A cross-sectional study method is being used to test the impact of exposure to raw wastewater and wastewater from storage reservoirs on diarrhoeal disease and parasitic infections in farmworkers and their families. The study population in the rainy season survey included 1 900 households: 680 households where the farmworker is exposed to untreated wastewater (exposed group), 520 households exposed to reservoir water (semi exposed group), and 700 households where the farmworker practices rain-fed agriculture (control group). Preliminary analysis of the data from the rainy season study (dry season study in progress) has been carried out. Current information indicates that the risk of Ascaris lumbricoides infection is much higher in the exposed group than in the semi-exposed and the control groups. According to the procedures employed Entamoeba histolytica infection was more frequent in individuals aged 5 to 14 years exposed to wastewater, than in those in the control group. When diarrhoeal disease rates were analyzed, children under age 15 years from exposed households had a significantly higher prevalence than controls. The final results of this study are expected to aid decisions on low cost treatment options and other health protection measures within the National Wastewater Reuse Programme in Mexico. The implications for assessment of the WHO guidelines for the use of wastewater in agriculture are discussed.     

Preliminary observations using a multi-layer ELISA method for the detection of Entamoeba histolytica trophozoite antigens in stool samples

A method for detecting Entamoeba histolytica trophozoite antigens in aqueous solution is described. This involves coating plastic microtitre plates with a 'catching antibody', specific rabbit anti-E. histolytica antibody (SRAE). After adding the test material the presence of antigen is determined using two additional heterologous antibody layers, one of 'developing antibody', in this case human anti-E. histolytica immunoglobulin (HAE), which is followed by a final layer of peroxidase conjugated sheep anti-human immunoglobulin antibody (SH-HRP). The specificity and sensitivity of the assay was investigated both in the model system and using stool samples from infected patients. In the model system, the test had a sensitivity equivalent to detection of approximately one amoeba per microscope coverslip (18 mm X 18 mm). Little cross reaction was observed with other intestinal parasites common to the area of Bangladesh from which the stool samples were taken. The possible use of this method in large scale screening of stool samples and in the detection of circulating antigens is discussed.     

Public health importance of non-biting cyclorrhaphan flies

This study was carried out to determine the role of non-biting cyclorrhaphan flies as carriers of human intestinal parasites at Woreta, northwestern Ethiopia. In total, 6530 flies were collected from four breeding sites and then examined for human intestinal parasites, mainly using the formol-ether concentration method. Fly species identified were Musca domestica (32.9%), Chrysomya rufifacies (32.6%), Musca sorbens (23%), Lucina cuprina (4.7%), Calliphora vicina (2.8%), Chrysomya bezziana (2.3%) and Wohlfahrtia magnifica (1.7%). Intestinal parasites such as Ascaris lumbricoides (36.9%), Trichuris trichiura (38.8%), hookworm (13.0%), Hymenolepis nana (0.6%), Taenia spp. (8.4%), Strongyloides stercoralis (1.7%), Entamoeba histolytica/dispar (48.1%), Entamoeba coli (24.7%), Cryptosporidium spp. (16.7%) and Giardia lamblia (10.4%) were isolated from both external and gut contents of the flies. Trichuris trichiura and A. lumbricoides among the helminths and E. histolytica/dispar and E. coli among the protozoans were the dominant parasites identified. It was observed that more parasites were isolated from gut contents than the external surfaces of the flies examined (P<0.001). Chrysomya rufifacies were found to carry more helminths than M. sorbens and M. domestica. Musca sorbens were the highest carriers of protozoan parasites followed by M. domestica and C. rufifacies. The significance of filth flies as carriers of human intestinal parasites has been highlighted.     

Prevalence of intestinal parasites in young Quichua children in the highlands of rural Ecuador

The prevalence of intestinal parasites in young Quichua children was assessed in 20 rural communities in the highlands of Ecuador in August 2005. The caregivers of 293 children aged 12-60 months were interviewed about the status of child health, household socioeconomic and environmental factors, and water-use practices and were requested to collect a faecal sample from the study child. Two hundred three (69.3%) of the 293 children provided faecal samples that were tested for parasites. The overall prevalences of infection for specific agents were Entamoeba histolytica or dispar 57.1%, Ascaris lumbricoides 35.5%, Entamoeba coli 34.0%, Giardia intestinalis (lamblia) 21.1%, Hymenolepis nana 11.3%, Cryptosporidium parvum 8.9%, Chilomastix mesnili 1.7%, Hymenolepis diminuta 1.0%, Strongyloides stercoralis 0.7%, and Trichuris trichiura 0.5%. The prevalence of parasites increased with age. Water storage, water treatment, consistent latrine-use, and participation in a community-based clean water project were not strongly associated with the prevalence of intestinal parasites, although having dirt floors was a risk factor for infection with E. histolytica or dispar and G. intestinalis.     

Intestinal parasitic infections and urbanization.

About a third of the population in the cities of developing countries live in slums and shanty towns. By the year 2000 it is estimated that this number will grow to 2200 million, and by 2025 about 57% of the population in developing countries will be in urban areas. The prevalence of infections caused by Entamoeba histolytica and Giardia intestinalis and the prevalence and intensity of Ascaris lumbricoides and Trichuris trichiura infections may increase among the rural populations who are migrating to these urban and suburban settings owing to the favourable conditions for transmission. Urgent consideration should therefore be given to improving sanitation in deprived urban areas and to treating periodically these populations to reduce the worm burden, especially in school-age children.     

Evaluation of uptake and generation of immune response by murine dendritic cells pulsed with hepatitis B surface antigen-loaded elastic liposomes

Hepatitis B surface antigen (HBsAg)-loaded elastic liposomes were studied for qualitative and quantitative uptake in vitro by murine dendritic cells (DCs) generated from bone marrow mononuclear cells. Internalization of the vesicles by the DCs was documented using fluorescence microscopy. Kinetics of uptake of antigen-loaded elastic vesicles by the DCs studied through flow cytometry showed a peak uptake at 6h. The ability of the antigen pulsed DCs to stimulate autologous peripheral blood lymphocytes was demonstrated by BrdU assay. Further evaluation by multiplex cytometric bead array analysis demonstrated a predominantly TH1 type of immune response. Our results suggest that HBsAg-loaded elastic vesicles as antigen delivery module and DCs as antigen presenting cells are able to generate a protective immune response. The property of elastic liposomes to traverse and target the immunological milieu of the skin makes it an attractive vehicle for development of a transcutaneous vaccine against hepatitis B virus.     

The epidemiology of infection with Entamoeba histolytica in the Gambia, West Africa.

A country-wide stool survey of The Gambia showed the prevalence of Entamoeba histolytica cysts to range from 13.7% up-country in the dry season to 52.3% near the coast. A longitudinal survey showed a near 100% infection rate over one year and a sharp rise in prevalence as the rains commence with an equally sharp fall as the rains progress. Specific antibody levels are elevated and reasonably constant through the year. Carriers generally show no specific lymphocyte reactivity to amoebic antigen but consistently parasite-negative individuals tended to show elevated lymphocyte reactivity. Attempts to discover the presence of cysts in the environment of villages by cultivation of specimens of water, soil, food, flies and washing from clothes and hands generally failed though E. histolytica was recovered once from a well.     

A comparison of skin-test responses using antigen from Leishmania donovani and a lizard trypanosome

261 individuals were skin tested with leishmanin from Leishmania donovani and with an antigen from a lizard trypanosome isolated in an area endemic for kala-azar in Ethiopia. 83 of the 261 responded positively, with indurations of 5 mm or greater, to one antigen but negatively to the other, and 42 reacted positively to both.Among the Afar people of the Awash valley in north-eastern Ethiopia and the Nyangatom people of south-western Ethiopia, positive response to both antigens was high, though many individuals responded to one antigen but not the other. Both these groups live as cattle pasturalists, a predominantly male occupation, in areas where known vectors of kala-azar as well as lizard-feeding sandflies exist. Among the Anuak and Suri people, who live in areas of south-western Ethiopia infested with tsetse flies, the lizard trypanosome antigen gave a markedly higher skin test positivity than did the leishmanin antigen. This suggests that the lizard trypanosome may give artificially high results in other immunological tests of exposure to trypanosomiasis in areas where lizard trypanosomes and man-biting sandflies co-exist with mammalian trypanosomes transmitted by Glossina.