Risk of acquiring Ascaris lumbricoides infection in an endemically infected rural community in Venezuela.

Soil-transmitted helminths, such as Ascaris lumbricoides, are the most prevalent parasites globally. Optimal anthelmintic treatment for A. lumbricoides in endemically infected communities is challenged by several host-related and environmental factors influencing infection acquisition. We assessed the risk of A. lumbricoides (re)infection after treatment in a Venezuelan rural community. Individual merthiolate-iodine-formaldehyde-fixed faecal samples were collected from 224 persons before a single-dose pyrantel treatment and at 1, 3, 6, 9 and 15 months after treatment. Effects of age, sex and socioeconomic status (SES) on A. lumbricoides prevalence, eggs/gram faeces (EPG) and infection (re)acquisition were assessed using both generalised linear mixed-effects models and survival analysis. Pre-treatment A. lumbricoides prevalence was 39.7%. Higher prevalence was associated with younger age and lower SES. Higher EPG values were observed among young children. Median time to A. lumbricoides infection was six months after treatment: at 1, 3, 6, 9 and 15 months post-treatment, cumulative incidence was 6.7%, 18.9%, 34.6%, 42.2%, and 52.6%, respectively. Younger age, lower SES, and pre-treatment A. lumbricoides infection status showed significantly elevated hazard ratios. Mass drug administration protocols would benefit from considering these factors in selective treatment strategies and possibly more than just annual or biannual treatments in the target population.

Hotspots and correlates of soil-transmitted helminth infections in a Venezuelan rural community: Which are the "wormy" houses?

OBJECTIVES: To determine spatial clustering and risk factors for occurrence and intensity of infection for soil-transmitted helminthiasis (STH), namely Ascaris lumbricoides, Trichuris trichiura, Strongyloides stercoralis and hookworms in a Venezuelan rural community. METHODS: MIF-fixed faecal samples were individually collected for STH testing. The Getis-Ord statistic was used to determine significant STH clustering within 25/50/100 m radiuses around houses. Individual- and house-level factors associated with STH occurrence and intensity of infection were determined using generalized estimating equations. RESULTS: Significant clusters of "wormy" houses for one or multiple parasites were found at distances of 25-50 m around 13 houses. Risk factors differed between occurrence and intensity of infection. Overcrowding in the house increased occurrence of S. stercoralis, T. trichiura and hookworm infections, while poor housing conditions increased A. lumbricoides infection risk. Overcrowding, poor faecal disposal system, economic dependency and lack of basic services differentially influenced the STHs. The "wormy" houses were mainly those built with waste materials, under economic dependency and lacking indoor water supply. CONCLUSIONS: STH distribution in a community is clustered, with significant hotspots of STH occurrence and intensity of infection and different associated risk factors. Targeting the "wormy" houses is expected to affect STH morbidity more efficiently.

Diagnostic performance of parasitological, immunological and molecular tests for the diagnosis of Schistosoma mansoni infection in a community of low transmission in Venezuela.

In Venezuela, areas endemic for schistosomiasis are of low transmission, with low parasite loads. Immunological tests often lack specificity and cannot differentiate past from present infections. Molecular tests are an alternative, although validation studies in endemic areas are needed. The aim of this study was to determine the performance of parasitological, immunological and molecular tests for the diagnosis of Schistosoma mansoni infection in low-transmission settings. A cross-sectional study was carried out in a rural community located in a schistosomiasis-endemic area of Venezuela to determine the prevalence and diagnostic performance of the Kato-Katz (KK) technique, Circumoval Precipitin Test (COPT), ELISA based on soluble egg antigen (ELISA-SEA) with and without treatment with sodium metaperiodate (ELISA-SEA-SMP), and PCR for amplification of the 121 bp highly repeated sequence of Schistosoma mansoni in faeces, urine and serum samples. The highest prevalence rates were obtained with ELISA-SEA (38.7%), COPT (33.3%), ELISA-SEA-SMP (31.5%), PCR on faeces (21.6%), and KK (17.1%), whereas PCR-based prevalence in urine was 6.2% and no positivity was detected in serum samples. Results showed that ELISA-SEA is the best method for the diagnosis of both current and former infections and that PCR on faeces is the best method for detecting recent transmission. The use of different tests that complement one another also allowed for a better diagnosis of Schistosoma mansoni infection, revealing a relatively high prevalence (33.8%) of schistosomiasis in a community of low transmission.

Case Report: Ocular Gnathostomiasis in Venezuela Most Likely Acquired in Texas.

A 69-year-old male dentist in Caracas, Venezuela, was referred to our Cornea Clinic with a history of pain, photophobia, and blurred vision on his left eye. Routine biomicroscopic examination with a slit lamp showed a worm in the corneal stroma of his left eye. The worm was surgically removed and was identified morphologically as Gnathostoma binucleatum.

Diagnosis of intestinal parasites in a rural community of Venezuela: Advantages and disadvantages of using microscopy or RT-PCR.

A cross-sectional study was carried out to determine the prevalence and diagnostic performance of microscopy and real time PCR (RT-PCR) for 14 intestinal parasites in a Venezuelan rural community with a long history of persistent intestinal parasitic infections despite the implementation of regular anthelminthic treatments. A total of 228 participants were included in this study. A multiplex RT-PCR was used for the detection of Dientamoeba fragilis, Giardia intestinalis, Cryptosporidium sp. and a monoplex RT-PCR for Entamoeba histolytica. Furthermore, a multiplex PCR was performed for detection of Ascaris lumbricoides, Strongyloides stercoralis, Necator americanus and Ancylostoma duodenale. Combined microscopy-PCR revealed prevalences of 49.3% for A. lumbricoides, 10.1% for N. americanus (no A. duodenale was detected), 2.0% for S. stercoralis, 40.4% for D. fragilis, 35.1% for G. intestinalis, and 7.9% for E. histolytica/dispar. Significant increases in prevalence at PCR vs. microscopy were found for A. lumbricoides, G. intestinalis and D. fragilis. Other parasites detected by microscopy alone were Trichuris trichiura (25.7%), Enterobius vermicularis (3.4%), Blastocystis sp. (65.8%), and the non-pathogenic Entamoeba coli (28.9%), Entamoeba hartmanni (12.3%), Endolimax nana (19.7%) and Iodamoeba bütschlii (7.5%). Age- but no gender-related differences in prevalences were found for A. lumbricoides, T. trichiura, G. intestinalis, and E. histolytica/dispar. The persistently high prevalences of intestinal helminths are probably related to the high faecal pollution as also evidenced by the high prevalences of non-pathogenic intestinal protozoans. These results highlight the importance of using sensitive diagnostic techniques in combination with microscopy to better estimate the prevalence of intestinal parasites, especially in the case of D. fragilis trophozoites, which deteriorate very rapidly and would be missed by microscopy. In addition, the differentiation between the pathogenic E. histolytica and the non-pathogenic E. dispar can be attained. However, microscopy remains an important diagnostic tool since it can detect other intestinal parasites for which no PCR is available.

Differential distribution and biochemical characteristics of hydrolases among developmental stages of Schistosoma mansoni may offer new anti-parasite targets.

Schistosoma mansoni enzymes play important roles in host-parasite interactions and are potential targets for immunological and/or pharmacological attack. The aim of this study was to comparatively assess the presence of hydrolytic activities (phosphatases, glycosidases, aminopeptidases) in soluble (SF) and membrane (MF) fractions from different S. mansoni developmental stages (schistosomula 0 and 3h, juveniles, and adult worms of 28 and 45days-old, respectively), by using simple enzyme-substrate microassays. Our results show and confirm the prominent presence of alkaline phosphatase (AlP) activity in the MF of all the above parasite stages, highlighting also the relevant presence of MF-associated α-mannosidase (α-MAN) activity in juveniles. A soluble AlP activity, together with ß-N-D-acetylglucosaminidase (ß-NAG), and α-MAN activities, was detected in SF of schistosomulum 0h. Soluble ß-NAG, α-MAN, acid phosphatase (AcP), leucin (LAP) and alanine (AAP) aminopeptidase activities were also seen in the SF of the other different developmental stages. This work shows different soluble and membrane-associated hydrolytic capacities in each S. mansoni developmental stage from schistosomula to adults that might be exploitable as potential new targets for immune and/or chemoprophylactic strategies.

Survival, induction and resuscitation of Vibrio cholerae from the viable but non-culturable state in the Southern Caribbean Sea.

The causative agent of cholera, Vibrio cholerae, can enter into a viable but non-culturable (VBNC) state in response to unfavorable conditions. The aim of this study was to evaluate the in situ survival of V. cholerae in an aquatic environment of the Southern Caribbean Sea, and its induction and resuscitation from the VBNC state. V. cholerae non-O1, non-O139 was inoculated into diffusion chambers placed at the Cuare Wildlife Refuge, Venezuela, and monitored for plate, total and viable cells counts. At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state. Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells. Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season. Resuscitation studies showed significant recovery of cell cultivability with spent media addition (p < 0.05). These results suggest that V. cholerae can persist in the VBNC state in this Caribbean environment and revert to a cultivable form under favorable conditions. The VBNC state might represent a critical step in cholera transmission in susceptible areas.

Survival, induction and resuscitation of vibrio cholerae from the viable but non-culturable state in the Southern Caribbean sea / Supervivencia, inducción y resucitación de Vibrio cholerae del estado viable no cultivable en el sur del Mar Caribe

The causative agent of cholera, Vibrio cholerae, can enter into a viable but non-culturable (VBNC) state in response to unfavorable conditions. The aim of this study was to evaluate the in situ survival of V. cholerae in an aquatic environment of the Southern Caribbean Sea, and its induction and resuscitation from the VBNC state. V. cholerae non-O1, non-O139 was inoculated into diffusion chambers placed at the Cuare Wildlife Refuge, Venezuela, and monitored for plate, total and viable cells counts. At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state. Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells. Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season. Resuscitation studies showed significant recovery of cell cultivability with spent media addition (p < 0.05). These results suggest that V. cholerae can persist in the VBNC state in this Caribbean environment and revert to a cultivable form under favorable conditions. The VBNC state might represent a critical step in cholera transmission in susceptible areas.

El agente causal del cólera, Vibrio cholerae, puede entrar a un estado viable no cultivable (VNC) en respuesta a condiciones desfavorables. El objetivo de este estudio fue evaluar la supervivencia in situ de V. cholerae en un ambiente acuático al sur del Mar Caribe y su inducción y resucitación del estado VBNC. V. cholerae no-O1, no-O139 fue inoculado en cámaras de difusión ubicadas en el Refugio de Fauna Cuare, Venezuela, y monitoreado para contaje de colonias, células totales y viables. En 119 días de exposición al ambiente, el contaje de colonias fue < 10 UFC/mL y una fracción de la población bacteriana entró al estado VBNC. Adicionalmente, la viabilidad disminuyó dos órdenes de magnitud y ocurrieron cambios morfológicos de células bacilares a cocoides. Entre las variables del ambiente acuático, la salinidad presentó correlación negativa con el contaje de colonias. Los estudios de resucitación mostraron recuperación significativa de la cultivabilidad celular con adición de sobrenadantes de cultivos en crecimiento activo (p < 0.05). Estos resultados sugieren que V. cholerae puede persistir en estado VBNC en este ambiente de Caribe y revertir a una forma cultivable bajo condiciones favorables. El estado VBNC podría representar un paso crítico en la transmisión del cólera en áreas susceptibles.

Are intestinal parasites fuelling the rise in dual burden households in Venezuela?

BACKGROUND: In developing countries undergoing rapid economic development, the number of dual burden households (i.e. co-existing overweight/obesity and stunting) is increasing. While intestinal parasites are prevalent in these countries, their contribution to dual burden households has so far been neglected. We studied the association between intestinal parasite infection and belonging to a dual burden household in a rural community of Venezuela. METHODS: We examined 225 individuals. A dual burden household was defined as a household with at least one overweight/obese adult (BMI > 25) and at least one stunted child (height -for-age z score <-2). Intestinal parasite (Giardia lamblia and geohelminth) infection was determined by faecal smears. RESULTS: In this community, 47.3% of the individuals were infected with intestinal parasites. Among adults, 65.2% were overweight/obese and 13.8% of the children were stunted. More than one in four households (26.8%) were dual burden households. Being infected with G. lamblia & geohelminths was significantly associated with being in a dual burden household (OR = 4.75, 95% CI: 1.01-22.20, n = 188), indicating a triple burden of disease in this community in Venezuela. CONCLUSION: While the relationship between intestinal parasite infection and stunting has been well established, these results indicate a need to further explore the association of intestinal parasite infection with dual burden households.

Household social determinants of ascariasis and trichuriasis in North Central Venezuela.

Ascariasis and trichuriasis are highly prevalent parasitoses in tropical countries caused by Ascaris lumbricoides and Trichuris trichiura, respectively. Their prevalence is associated with poor sanitation and hygiene, among other biological and social factors. In this study, the associations of household social determinants with these soil-transmitted helminthiases in North Central Venezuela was assessed in the context of a National Study on Human Growth and Development (SENACREDH). This was a probabilistic, stratified, cross-sectional survey with a sample of 3388 individuals that weighted represents 4 675 433 inhabitants from the region. The prevalence of ascariasis and trichuriasis was 3.73% and 1.13%, respectively. In the univariate and multivariate analyses, these prevalences were significantly higher in those living in vulnerable houses [adjusted OR (AOR) = 1.479, 95% CI 1.428-1.532; and AOR = 10.519, 95% CI 9.971-11.097, respectively], houses located in rural areas (AOR = 2.067, 95% CI 2.035-2.101; and AOR = 1.918, 95% CI 1.868-1.970, respectively) and houses with a soil floor (AOR = 5.027, 95% CI 4.895-5.162; and AOR = 5.190, 95% CI 4.944-5.448, respectively), among other factors. People living in rural households with inappropriate building materials, insufficient basic public services such as water and waste disposal, and surrounding disease-prone environments are at higher risk of acquiring A. lumbricoides and T. trichiura infections.

Hyperinfection by Strongyloides stercoralis probably associated with Rituximab in a patient with mantle cell lymphoma and hyper eosinophilia / Hiperinfección por Strongyloides stercoralis probablemente asociada con Rituximab en una paciente con linfoma e hipereosinofilia

The first report to our knowledge, of hyperinfection by Strongyloides stercoralis (HS) and hypereosinophilia, associated to immune suppression by Rituximab (the only drug received for the last one year and 10 months), in a patient with mantle-cell lymphoma (MCL), is presented. The patient has a 3-year history of MCL, and developed two accesses of HS during 2008, including meningitis, pneumonia and presence of larvae of S. stercoralis in the lungs. We had a unique chance to look at cytotoxicity of filariform larvae in the expectoration after Ivermectin treatment, showing immobilization and death of larvae, associated with eosinophils attached to the cuticle of the parasite.

Se presenta el primer reporte, hasta donde tengamos información, de hiperinfección por Strongyloides stercoralis (HS) e hipereosinofilia asociados a inmunosupresión por Rituximab (el único medicamento recibido durante 1 año y 10 meses), en un paciente con linfoma de células del manto (LCM). La paciente tuvo una historia de 3 años con LCM, y desarrolló 2 accesos de HS durante el 2008, incluyendo meningitis, neumonía y presencia de larvas de S. stercoralis en los pulmones. Se tuvo la oportunidad única de observar la citotoxicidad contra las larvas filariformes en la expectoración, luego del tratamiento con Ivermectina, mostrando la inmovilización y muerte de las larvas, asociada a la presencia de eosinófilos adheridos a la cutícula del parásito.

Staphylococcus warneri meningitis in a patient with Strongyloides stercoralis hyperinfection and lymphoma: first report of a case.

A case of meningitis due to Staphylococcus warneri in a patient with a hyperinfection with Strongyloides stercoralis possibly associated with rituximab treatment for mantel cell lymphoma is reported for the first time in the literature. The patient was a 59-year-old woman, with a 3-year history of an apparently well controlled lymphoma after treatment with chemotherapy-immunotherapy and then immunotherapy alone, and diagnosis of strongyloidiasis. Meningitis was diagnosed by cerebrospinal fluid culture and tested with an automated plate system. The patient was successfully treated with vancomycin; although fever and productive cough persisted. Severe gastrointestinal symptoms and pneumonia developed three weeks later. Hyperinfection syndrome by S. stercoralis was diagnosed, with abundant larvae in feces and expectoration.

Staphylococcus warneri meningitis in a patient with Strongyloides stercoralis hyperinfection and lymphoma: first report of a case / Staphylococcus warneri meningite em paciente com hiperinfecção com Strongyloides stercoralis e linfoma: primeiro relato de caso

A case of meningitis due to Staphylococcus warneri in a patient with a hyperinfection with Strongyloides stercoralis possibly associated with rituximab treatment for mantel cell lymphoma is reported for the first time in the literature. The patient was a 59-year-old woman, with a 3-year history of an apparently well controlled lymphoma after treatment with chemotherapy-immunotherapy and then immunotherapy alone, and diagnosis of strongyloidiasis. Meningitis was diagnosed by cerebrospinal fluid culture and tested with an automated plate system. The patient was successfully treated with vancomycin; although fever and productive cough persisted. Severe gastrointestinal symptoms and pneumonia developed three weeks later. Hyperinfection syndrome by S. stercoralis was diagnosed, with abundant larvae in feces and expectoration.

Caso de meningite por Staphylococcus warneri em paciente com hiperinfecção com Strongyloides stercoralis, possivelmente associada com tratamento de rituximab para linfoma de células do manto é relatada pela primeira vez na literatura. A paciente, mulher de 59 anos com história de três anos de linfoma aparentemente bem controlado com tratamento com quimioterapia-imunoterapia e, em seguida, somente imunoterapia e diagnóstico de estrongiloidíase. Meningite foi diagnosticada por cultura do líquido cefalorraquidiano e testada com sistema automatizado de placa. A paciente foi tratada com sucesso com vancomicina, embora a febre e a tosse produtiva não tenham desaparecido. Após graves sintomas gastrointestinais a paciente desenvolveu pneumonia três semanas mais tarde. Síndrome de hiperinfecção por S. stercoralis foi diagnosticada, com larvas abundantes nas fezes e expectoração.

Hyperinfection by Strongyloides stercoralis probably associated with Rituximab in a patient with mantle cell lymphoma and hyper eosinophilia.

The first report to our knowledge, of hyperinfection by Strongyloides stercoralis (HS) and hypereosinophilia, associated to immune suppression by Rituximab (the only drug received for the last one year and 10 months), in a patient with mantle-cell lymphoma (MCL), is presented. The patient has a 3-year history of MCL, and developed two accesses of HS during 2008, including meningitis, pneumonia and presence of larvae of S. stercoralis in the lungs. We had a unique chance to look at cytotoxicity of filariform larvae in the expectoration after Ivermectin treatment, showing immobilization and death of larvae, associated with eosinophils attached to the cuticle of the parasite.

Human infection by Angiostrongylus costaricensis in Venezuela: first report of a confirmed case.

A proven case of human infection caused by Angiostrongylus costaricensis is reported for the first time in Venezuela. The patient was a 57-year-old female surgically operated because of signs of peritonitis with a palpable mass at the lower right quadrant of the abdomen. WBC count reported 16,600 cells/mm3, with 46% eosinophils. The tumoral aspect of ileocolic area and peritoneal lymph nodes prompted the resection of a large area of the terminal ileum, cecum, part of the ascending colon and a small part of the jejunum, where a small lesion was found. The pathology showed thickened areas of the intestinal wall with areas of hemorrhage and a perforation of the cecum. Histology showed intense eosinophil infiltration of the whole intestinal wall, granulomas with giant cells and eosinophils. Some of the granuloma surrounded round or oval eggs with content characterized by a large empty area, cells or embryo in the center, and sometimes nematode larvae. A cross section of an adult nematode worm was observed inside a branch of mesenteric artery. The intestinal affected area, the characteristics of the lesions, the presence of eggs in the submucosa with nematode larvae inside, and the observation of a nematode inside a mesenteric artery, makes sufficient criteria for the diagnosis of an infection by Angiostrongylus costaricensis.

Human infection by Angiostrongylus costaricensis in Venezuela: first report of a confirmed case

A proven case of human infection caused by Angiostrongylus costaricensis is reported for the first time in Venezuela. The patient was a 57-year-old female surgically operated because of signs of peritonitis with a palpable mass at the lower right quadrant of the abdomen. WBC count reported 16,600 cells/mm³, with 46 percent eosinophils. The tumoral aspect of ileocolic area and peritoneal lymph nodes prompted the resection of a large area of the terminal ileum, cecum, part of the ascending colon and a small part of the jejunum, where a small lesion was found. The pathology showed thickened areas of the intestinal wall with areas of hemorrhage and a perforation of the cecum. Histology showed intense eosinophil infiltration of the whole intestinal wall, granulomas with giant cells and eosinophils. Some of the granuloma surrounded round or oval eggs with content characterized by a large empty area, cells or embryo in the center, and sometimes nematode larvae. A cross section of an adult nematode worm was observed inside a branch of mesenteric artery. The intestinal affected area, the characteristics of the lesions, the presence of eggs in the submucosa with nematode larvae inside, and the observation of a nematode inside a mesenteric artery, makes sufficient criteria for the diagnosis of an infection by Angiostrongylus costaricensis.

Un caso comprobado de infección humana por Angiostrongylus costaricensis es reportado para la literatura por vez primera en Venezuela. El paciente fue una mujer de 57 años de edad intervenida quirúrgicamente debido a signos de peritonitis, con una masa palpable en la fosa ilíaca derecha del abdomen. El recuento de leucocitos reportó 16.600 células/mm³ con 46 por ciento eosinófilos. El aspecto tumoral del área íleocólica y las adenopatías peritoneales encontradas motivó la resección de un área grande del ileon terminal, ciego, parte del colon ascendente y una pequeña parte del yeyuno donde se encontró una pequeña lesión interpretada como metastásica. La patología mostró áreas engrosadas de la pared intestinal con zonas de hemorragia y perforación del ciego. El estudio histológico mostró una intensa infiltración eosinofílica de toda la pared intestinal, granulomas con células gigantes y eosinófilos. Algunos de los granulomas rodeaban huevos ovales o redondos con un contenido caracterizado por una gran área vacía, células o embriones en el centro, y en algunos casos una larva de nematodo. Un corte transversal de un verme nematodo adulto se observó dentro de una arteria mesentérica. El área intestinal afectada, las características de las lesiones, la presencia de huevos en la submucosa con larvas de nematodo dentro, y la observación de un nematodo dentro de una arteria mesentérica, aportan suficientes criterios para el diagnóstico de una infección por Angiostrongylus costaricensis.

Preliminary antigenic characterisation of an adult worm vomit preparation of Fasciola hepatica by infected human sera.

Fascioliasis is an emerging/re-emerging vector-borne disease with the widest known distribution. Approximately 17 million people are infected around the world, being the Andean region the most affected area. There is an important necessity to develop sensitive and specific diagnostic tools to treat patients early and to avoid complications. In this paper we evaluated the immune response of infected humans against two antigenic preparations: the total soluble extract (FhTSE) and the adult worm vomit (FhAWV) in order to identify antigenic fractions specific for Fasciola hepatica. Both preparations were processed by SDS-PAGE and Western blot with human sera with fascioliasis (F), other parasitosis and healthy individuals. In the immunoblot of FhTSE, sera F recognised 16 bands with MW between eight and 110 kDa, from which those of 8, 9, 10, 38, 45 and 57 kDa were specific. In the preparation FhAWV, sera F recognised nine bands with MW from eight to 85 kDa, from which those of 8, 12, 15 and 24 kDa were specific. Some bands of cross-reaction were evident with sera from patients with other parasitoses, more frequent with the FhTSE. Bands within the MW mentioned, particularly that of eight kDa, have been shown to be specific by others, and deserve additional characterisation for their potential use in immunodiagnosis.

Preliminary characterization of an adult worm "vomit" preparation of Schistosoma mansoni and its potential use as antigen for diagnosis.

Schistosoma mansoni is a parasitic trematode of the portal-mesenteric veins with a closed-end intestine. Adult worms regurgitate their intestinal content after digestion, together with constituents of the lining gut. Some of these molecules circulate in the blood and are antigenic. We obtain a "vomit" preparation and preliminary evaluate its biochemical composition and antigenic capacity. The "vomit" preparation was obtained after changes in temperature and solutions of incubation of adult worms between 4 and 37 degrees C. Supernatant was assayed for protein, carbohydrate concentration and enzymatic activities associated to the intestine and to the worm tegument. The antigenicity of the product was evaluated using Western blot (WB) analysis against sera of experimentally infected mice, before and after drug cure, sera from people infected with S. mansoni and from individuals infected with other parasitoses. More carbohydrate than protein was detected in the preparations. Cysteine proteinase (CP), N-acetyl-beta-D: -glucosaminidase and alkaline phosphatase activities were detected. The latter enzyme activity is a marker of the tegument, suggesting that in spite of careful conditions used to avoid the presence of tegumental material, manipulation of the worms always resulted in the release of tegumental molecules. Cationic exchange chromatography was useful to separate various components of this "vomit" preparation, particularly enzymes responsible for CP activity. Two highly immunogenic and specific duplets were observed in the WB analysis, 31/32- and 38/40-kDa components, the former probably referring to the intestinal CPs Sm31/Sm32. None of the two duplets disappeared after successful chemotherapy during the time of evaluation in mice or humans.

Preliminary antigenic characterisation of an adult worm vomit preparation of Fasciola hepatica by infected human sera

Fascioliasis is an emerging/re-emerging vector-borne disease with the widest known distribution. Approximately 17 million people are infected around the world, being the Andean region the most affected area. There is an important necessity to develop sensitive and specific diagnostic tools to treat patients early and to avoid complications. In this paper we evaluated the immune response of infected humans against two antigenic preparations: the total soluble extract (FhTSE) and the adult worm vomit (FhAWV) in order to identify antigenic fractions specific for Fasciola hepatica. Both preparations were processed by SDS-PAGE and Western blot with human sera with fascioliasis (F), other parasitosis and healthy individuals. In the immunoblot of FhTSE, sera F recognised 16 bands with MW between eight and 110 kDa, from which those of 8, 9, 10, 38, 45 and 57 kDa were specific. In the preparation FhAWV, sera F recognised nine bands with MW from eight to 85 kDa, from which those of 8, 12, 15 and 24 kDa were specific. Some bands of cross-reaction were evident with sera from patients with other parasitoses, more frequent with the FhTSE. Bands within the MW mentioned, particularly that of eight kDa, have been shown to be specific by others, and deserve additional characterisation for their potential use in immunodiagnosis.

Fasciolíase é uma doença emergente/re-emergente transmitida por vetores com a distribuição sabidamente mais ampla. Existem aproximadamente 17 milhões de pessoas infectadas em todo mundo, sendo a região andina a área mais afetada. Há uma necessidade importante para desenvolver ferramentas diagnósticas sensíveis e específicas para tratar cedo os pacientes e para evitar complicações. Neste trabalho avaliamos a resposta imune de seres humanos infectados comparando a duas preparações antigênicas: o extrato solúvel total (FhTSE) e o vômito (FhAWV) do verme adulto a fim de identificar as frações antigênicas específicas para Fasciola hepatica. Ambas as preparações foram processadas por SDS-PAGE e Western blot com os soros humanos de portadores de fasciolíase (F), outras parasitoses e indivíduos saudáveis. No immunoblot de FhTSE, os soros F reconheceram 16 faixas com PM entre 8 e 110 kDa, das quais as de 8, 9, 10, 38, 45 e 57 kDa foram específicas. Na preparação de FhAWV, os soros F reconheceram 9 faixas com PM entre 8 e 85 kDa, das quais as de 8, 12, 15 e 24 kDa foram específicas. Algumas faixas com reação cruzada foram evidentes com os soros dos pacientes com outras parasitoses, mais freqüentes com o FhTSE. As faixas dentro do PM mencionado, particularmente aquela de 8 kDa, mostraram ser específicas por outros autores, e merecem a caracterização adicional para seu uso potencial no diagnóstico imunológico.