Toxocariasis, Intestinal Parasitoses and Eosinophilia in Schoolchildren from Argentina.

The purpose was to analyze the relationship between toxocariasis, intestinal parasitosis, eosinophilia, pet ownership and signs and symptoms present in 370 schoolchildren (3-12 years old) of Argentina. The positive serology for toxocariasis was 19.5%, associated with eosinophilia and possession of cats as pets. The stool testing revealed 70.0% of parasitized children. Its frequency was 44.9% helminthes and 53.5% protozoan resulting single significant presence of protozoa in children aged 10-12 years. The species were: Blastocystis hominis (45.7%), Enterobius vermicularis (43.2%), Giardia intestinalis (16.2%), Hymenolepis nana (2.4%), Ascaris lumbridoides (1.6%) and Trichuris trichiura (0.3%). The survey showed 13.0% of children with sporadic abdominal pain and 13.5% with pruritus ani. The results indicate relationship between positive serology and eosinophilia compatible with a clinical aspect of covert toxocariasis self-limiting, associated with cats ownership; in school children from our region who also have high frequency of intestinal parasitoses.

Distribution of intestinal parasitoses in relation to environmental and sociocultural parameters in La Plata, Argentina.

Environmental parameters influencing the distribution of parasite species in three neighbourhoods of differing socioeconomic conditions in La Plata, Argentina were analysed. Coproparasitological screenings were performed in children up to 14 years old from a marginal zone (100), a suburban neighbourhood (101), and an urban area (91) in 1999--2000. The presence of parasite species in environmental samples (water and soil) and the degree of association among parasite communities was documented and evaluated. The prevalence of infection in each population was 73.0%, 54.4% and 35.2%, respectively. The frequencies of helminths and pathogenic protozoa were both higher in the marginal zone, where sanitary and environmental conditions were significantly inferior compared with the other zones. The high prevalence of intestinal parasites in this infantile population was related to parasitic contamination of the soil and water sources in addition to deficient sanitary and sociocultural conditions. Calculation of an equitability index revealed that the specific richness was less equitable once socioeconomic conditions and hygienic practices were improved. This study demonstrates the need to implement management practices for the control of intestinal parasitoses in accordance with the environmental and sociocultural characteristics of a given ecosystem.

Resistance to antibiotics in injured coliforms isolated from drinking water.

We studied the antibiotic sensitivity of injured coliforms isolated from drinking water of La Plata, Argentina. The antibiotic sensitivity test by the agar diffusion method were proved in: Klebsiella oxytoca (14 strains), Enterobacter aerogenes (4 strains) and Enterobacter cloacae genomic group 3 (14 strains). We found that while these impaired total coliforms were sensitive to piperacillin-tazobactam (TAZ), netilmicin (NTL), ofloxacin (OFLX), and norfloxacin (NFLX) (100%), they had resistant to aminopenicillin-sulbactam (AMS) and nitrofurantoin (NIT) (100%). The resistance to antibiotics demonstrated in these strains would point to the need to promote a rational and judicious use of antimicrobial agents while at the same time implementing a program of active vigilance aimed at ensuring the highest quality of drinking water throughout the system.

[Isolation and characterization of injured coliforms from the drinking water distribution network of La Plata, Argentina]. / Aislamiento y caracterización de coliformes injuriados provenientes de la red de distribución de agua de bebida de La Plata, Argentina.

We screened the La Plata drinking water distribution network for fecal and total coliform bacterial indicator by purification procedures, cultivating 66 membrane-filtered samples from the two networks on m-T7 agar. Subterranean and river-derived water yielded 13 and 18 confirmed gram-negative bacillus isolates, with 54% and 72% representing total coliforms, respectively. Those from the former source were Klebsiella oxytoca, Enterobacter agglomerans, and Enterobacter aerogenes and from the latter Klebsiella oxytoca, Enterobacter agglomerans, and Enterobacter cloacae, genomic group 3. Since 58% of the samples were positive using m-T7 medium it is suggested that the inclusion in standard quality control protocols should be implemented.

Aislamiento y caracterización de coliformes injuriados provenientes de la red de distribución de agua de bebida de La Plata, Argentina / Isolation and characterization of injured coliforms from the drinking water distribution network of La Plata, Argentina

We screened the La Plata drinking water distribution network for fecal and total coliform bacterial indicator by purification procedures, cultivating 66 membrane-filtered samples from the two networks on m-T7 agar. Subterranean and river-derived water yielded 13 and 18 confirmed gram-negative bacillus isolates, with 54 and 72 representing total coliforms, respectively. Those from the former source were Klebsiella oxytoca, Enterobacter agglomerans, and Enterobacter aerogenes and from the latter Klebsiella oxytoca, Enterobacter agglomerans, and Enterobacter cloacae, genomic group 3. Since 58 of the samples were positive using m-T7 medium it is suggested that the inclusion in standard quality control protocols should be implemented.

Aislamiento y caracterización de coliformes injuriados provenientes de la red de distribución de agua de bebida de La Plata, Argentina / Isolation and characterization of injured coliforms from the drinking water distribution network of La Plata, Argentina

We screened the La Plata drinking water distribution network for fecal and total coliform bacterial indicator by purification procedures, cultivating 66 membrane-filtered samples from the two networks on m-T7 agar. Subterranean and river-derived water yielded 13 and 18 confirmed gram-negative bacillus isolates, with 54 and 72 representing total coliforms, respectively. Those from the former source were Klebsiella oxytoca, Enterobacter agglomerans, and Enterobacter aerogenes and from the latter Klebsiella oxytoca, Enterobacter agglomerans, and Enterobacter cloacae, genomic group 3. Since 58 of the samples were positive using m-T7 medium it is suggested that the inclusion in standard quality control protocols should be implemented.(AU)

Intestinal parasites and the environment: frequency of intestinal parasites in children of Neuquén, Patagonia, Argentina.

The frequency of intestinal parasites (IP) in 69 children between 2 and 14 years of age was studied within the city of Neuquén, Patagonia, Argentina in relation to socioeconomic, cultural and environmental conditions. Soil type was determined and contamination with IP was investigated in 75 soil samples. Frequency of IP in children was 50.7%. Blastocystis hominis was the most frequent species (24.6%). Simultaneous infection with 2 to 4 species was found in 47,5% of the infected children. Statistical association was found between parasitic infection and family hygienic level (p < 0.001). Socioeconomic level was defined as lower middle class, however, infrastructure and public services were adequate. The climate is continental semiarid and the soil is sandy with a low organic matter level. Intestinal protozoan organisms were found in 33.3% of soil samples and Sarcocystis sp. was the most frequent species (14.6%). The parasites present in soil are related to the environmental conditions that allow their viability and development. Socioeconomic and cultural factors are important for the appearance and spread of IP in communities where sanitary conditions and infrastructure are adequate.

Aislamiento y caracterización de coliformes injuriados provenientes de la red de distribución de agua de bebida de La Plata, Argentina. / [Isolation and characterization of injured coliforms from the drinking water distribution network of La Plata, Argentina]

We screened the La Plata drinking water distribution network for fecal and total coliform bacterial indicator by purification procedures, cultivating 66 membrane-filtered samples from the two networks on m-T7 agar. Subterranean and river-derived water yielded 13 and 18 confirmed gram-negative bacillus isolates, with 54

and 72

representing total coliforms, respectively. Those from the former source were Klebsiella oxytoca, Enterobacter agglomerans, and Enterobacter aerogenes and from the latter Klebsiella oxytoca, Enterobacter agglomerans, and Enterobacter cloacae, genomic group 3. Since 58

of the samples were positive using m-T7 medium it is suggested that the inclusion in standard quality control protocols should be implemented.

Relación entre el gradiente discontinuo de sacarosa y la viabilidad de ooquistes de Cryptosporidium sp / Relationship between discontinuous sucrose gradient and viability of Cryptosporidium sp. oocysts

Cryptosporidium sp. causes self-limiting gastroenteritis in immunocompetent individuals and can produce a life-threatening chronic diarrhea in immunodeficient patients. In order to obtain populations of selectively infective oocysts for inoculation in biological experiments, we developed an operational protocol for the enrichment of viable oocysts from crude fecal material. Using either fresh or formaldehyde-fixed feces as sources of viable and nonviable oocysts, respectively, we fractionated the samples on parallel discontinuous sucrose gradients and evaluated oocyst viability at different banding densities by an in vitro excystation assay. The formaldehyde-inactivated fecal samples formed no bands after centrifugation and 91.66 of the oocysts became concentrated in the pellet. Fresh fecal samples formed three bands at densities 1.062, 1.092, and 1.121, in addition to a sediment. Here the viability of the gradient-sedimented oocysts was 92.3 overall, and of those in the second band 100. Modifications in oocyst permeability thus seems to alter their sedimentation characteristics so that consequent distribution on sucrose gradients can be correlated with oocyst viability. Discontinuous sucrose density gradient sedimentation would thus constitute a simple and rapid mean to obtain viable oocysts for use in biological models both in vivo and in vitro.

Relación entre el gradiente discontinuo de sacarosa y la viabilidad de ooquistes de Cryptosporidium sp / Relationship between discontinuous sucrose gradient and viability of Cryptosporidium sp. oocysts

Cryptosporidium sp. causes self-limiting gastroenteritis in immunocompetent individuals and can produce a life-threatening chronic diarrhea in immunodeficient patients. In order to obtain populations of selectively infective oocysts for inoculation in biological experiments, we developed an operational protocol for the enrichment of viable oocysts from crude fecal material. Using either fresh or formaldehyde-fixed feces as sources of viable and nonviable oocysts, respectively, we fractionated the samples on parallel discontinuous sucrose gradients and evaluated oocyst viability at different banding densities by an in vitro excystation assay. The formaldehyde-inactivated fecal samples formed no bands after centrifugation and 91.66 of the oocysts became concentrated in the pellet. Fresh fecal samples formed three bands at densities 1.062, 1.092, and 1.121, in addition to a sediment. Here the viability of the gradient-sedimented oocysts was 92.3 overall, and of those in the second band 100. Modifications in oocyst permeability thus seems to alter their sedimentation characteristics so that consequent distribution on sucrose gradients can be correlated with oocyst viability. Discontinuous sucrose density gradient sedimentation would thus constitute a simple and rapid mean to obtain viable oocysts for use in biological models both in vivo and in vitro.(AU)

[Relationship between discontinuous sucrose gradient and viability of Cryptosporidium sp. oocysts]. / Relación entre el gradiente discontinuo de sacarosa y la viabilidad de ooquistes de Cryptosporidium sp.

Cryptosporidium sp. causes self-limiting gastroenteritis in immunocompetent individuals and can produce a life-threatening chronic diarrhea in immunodeficient patients. In order to obtain populations of selectively infective oocysts for inoculation in biological experiments, we developed an operational protocol for the enrichment of viable oocysts from crude fecal material. Using either fresh or formaldehyde-fixed feces as sources of viable and nonviable oocysts, respectively, we fractionated the samples on parallel discontinuous sucrose gradients and evaluated oocyst viability at different banding densities by an in vitro excystation assay. The formaldehyde-inactivated fecal samples formed no bands after centrifugation and 91.66% of the oocysts became concentrated in the pellet. Fresh fecal samples formed three bands at densities 1.062, 1.092, and 1.121, in addition to a sediment. Here the viability of the gradient-sedimented oocysts was 92.3% overall, and of those in the second band 100%. Modifications in oocyst permeability thus seems to alter their sedimentation characteristics so that consequent distribution on sucrose gradients can be correlated with oocyst viability. Discontinuous sucrose density gradient sedimentation would thus constitute a simple and rapid mean to obtain viable oocysts for use in biological models both in vivo and in vitro.

[Factors affecting the in vitro excystation of Cryptosporidium sp]. / Factores que influyen en el desenquistamiento in vitro de Cryptosporidium sp.

To assess the viability of the oocysts from Cryptosporidium sp, an in vitro excystation technique was developed and optimized. To this end, oocysts from Cryptosporidium sp were isolated from a sample of fecal material from a child with symptomatic cryptosporidiosis. The specimen was processed by a discontinuous-sucrose-gradient technique yielding a final suspension of purified and concentrated oocysts. The resulting oocysts were quantified by counting an aliquot in a Neubauer hemocytometer chamber and the remainder were preserved in phosphate-buffered saline at pH 7.2 and 4 degrees C. The two protocols tested through the use of 100-microliter aliquots containing 178,000 oocytes/mm3 were: I. Treatment at 37 degrees C for 24 h with 1% (v/v) bile plus 0.44% (w/v) sodium bicarbonate. Group A: pretreated with acidified trypsin (pH 2.75) at 37 degrees C for 1 h; Group B: not pretreated. II. Treatment at 37 degrees C for 24 h with 1% (v/v) bile at pH 6 and at pH 8 either with or without 0.44% (w/v) sodium bicarbonate in air or an atmosphere containing 10% (v/v) CO2. Group 1: bile, pH 6; Group 2: bile, pH 6 plus CO2; Group 3: bile, pH 6 plus bicarbonate; Group 4: bile, pH 8 plus bicarbonate; Group 5: bile, pH 6 plus bicarbonate plus CO2; Group 6: bile, pH 8 plus bicarbonate plus CO2; Group 7: bile, pH 8; and Group 8: bile, pH 8 plus CO2. We then assessed the degree of excystation in all the groups by counting the number of intact oocysts in the hemocytometer. For the first protocol, the oocyst count after excystation revealed an average of 28,000 oocysts/mm3 in Group A and 22,000 in Group B. These values indicate excystation frequencies of 84.3% and 87.6% for these two groups, respectively. The corresponding excystation percentages for groups 1-8 of protocol II were 49.5, 56.7, 83.2, 88.3, 88.3, 92.7, and 94.1, respectively. Preincubation with trypsin failed to augment oocyst excystation in Cryptosporidium in any of these experimental circumstances. The optimal conditions for oocyst excystation in these 24 h incubations were thus seen to be in the presence of 1% (v/v) bile at pH 8, and 37 degrees C under an atmosphere of 10% (v/v) CO2.

Factores que influyen en el desenquistamiento in vitro de Cryptosporidium sp / Factors affecting the in vitro excystation of Cryptosporidium sp

Para evaluar la viabilidad de los ooquistes de Cryptosporidium sp se desarrolló y optimizó una técnica de desenquistamiento in vitro. Los ooquistes de Cryptosporidium sp se obtuvieron de una muestra de materia fecal de un niño con cryptosporidiosis sintomática. La muestra fue procesada por la técnica de gradiente discontinuo de sacarosa hasta obtener una suspensión de ooquistes purificados y concentrados (OPC). Los ooquistes se cuantificaron en cámara de Neubauer y preservaron en PBS pH 7, 2 a 4§C. Los dos protocolos ensayados a partir de alícuotas de 100µl de OPC con 178.000 ooquistes/mm3 fueron: I. Tratamiento con bilis 1 por ciento y bicarbonato de sodio al 0,44 por ciento a 37§C durante 24 horas. II Tratamiento con bilis 1 por ciento con pH 6 y pH 8, con y sin bicarbonato de sodio al 0,44 por ciento, con y sin incubación en atmósfera de 10 por ciento de CO2 durante 24 h a 37§C. Se evaluó el grado de desenquistamiento en todos los grupos efectuando el conteo de ooquistes intactos en cámara de Neubauer. La preincubación con tripsina no incrementó el desenquistamiento de los ooquistes de Cryptosporidium sp. Las mejores condiciones de desenquistamiento fueron utilizando bilis al 1 por ciento pH 8, incubado con 10 por ciento de CO2, a 37§C durante 24 h.

Factores que influyen en el desenquistamiento in vitro de Cryptosporidium sp / Factors affecting the in vitro excystation of Cryptosporidium sp

Para evaluar la viabilidad de los ooquistes de Cryptosporidium sp se desarrolló y optimizó una técnica de desenquistamiento in vitro. Los ooquistes de Cryptosporidium sp se obtuvieron de una muestra de materia fecal de un niño con cryptosporidiosis sintomática. La muestra fue procesada por la técnica de gradiente discontinuo de sacarosa hasta obtener una suspensión de ooquistes purificados y concentrados (OPC). Los ooquistes se cuantificaron en cámara de Neubauer y preservaron en PBS pH 7, 2 a 4ºC. Los dos protocolos ensayados a partir de alícuotas de 100Al de OPC con 178.000 ooquistes/mm3 fueron: I. Tratamiento con bilis 1 por ciento y bicarbonato de sodio al 0,44 por ciento a 37ºC durante 24 horas. II Tratamiento con bilis 1 por ciento con pH 6

Characterization and quantitation of parasite species in the effluents of the Berisso main sewage channel, Buenos Aires, Argentina.

Outbreaks of disease can be caused by pathogenic intestinal parasites in drinking water. La Plata, the capital of the province of Buenos Aires, Argentina, is located on the La Plata River; a principal tributary receiving the raw sewage from the city's 500,000 inhabitants via the Berisso main septic channel at an estimated volume of 108,390 m3/day (4516.3m3/h). To determine the parasite-contamination level of the La Plata River, we obtained 18 sewage effluents from the Berisso main septic channel at its point of discharge into the river and concentrated these samples by filtration. This approach detected the following parasites at the indicated average levels (cysts/m3): members of the flagellate genus Giardia, more than 9000; representatives of the amoebic species Entamoeba histolytica, slightly more than 800 and Entamoeba coli, 5000; other amoebic forms, up to 23,000. These data clearly illustrate the degree of contamination within the La Plata River caused by the Berisso septic channel. Pollution of the La Plata River to this extent thus presents a high health risk for people who drink the water from this river even after conventional processing as well as for those who use the water from this portion of the river for recreation.

[Intestinal parasite infections in a periurban community from the Province of Buenos Aires, Argentina]. / Estudio de las infecciones por enteroparásitos en una comunidad periurbana de la provincia de Buenos Aires, Argentina.

A survey for intestinal parasites was performed on 38 individuals within the urban area of La Plata City (Province of Buenos Aires, Argentina). This community is composed of brick-factory workers who also live in the factory premises. An analysis for intestinal parasites was done on fecal samples collected serially and by means of anal swabs and thereafter preserved in formol solution. At the same time, the occurrence of the parasites under study as well as that of commensal organisms was investigated in water and soil samples within the factory environs. Information was also obtained from the members of this community as their age, sex, birthplace, and recent travels, either abroad or to the interior of Argentina. The prevalence of the pathogens and commensal parasites was 89.5%. The frequency of protozoans and helminths was: G. lamblia 26.3%, B. hominis 65.8%, A. lumbricoides 7.9%, H. nana 2.6%, Uncinaria sp. 7.9%, S. stercoralis 2.6% and E. vermicularis 42.1%. None of these parasite or commensal organisms were present in the water samples investigated. Four out of 20 soil samples analyzed contained parasitic elements: T. canis eggs (one), G. lamblia cysts (two) and A. lumbricoides eggs (one). These results indicated that most important factors causing such a high prevalence of coproparasites were the poor conditions of personal and community hygiene in combination with the frequent travels to the north and the northeast of the country, regions which are endemic parasitic areas. The implementation of programs on health education and communal sanitation would contribute in the control of this health problem.

Relationship between parasitical inoculum and immunological parameters in experimental toxocariasis.

Swiss female mice were inoculated with different infective doses of Toxocara canis eggs to evaluate the percentage of recuperated larvae in different organs and immunological parameters such as spleen index (SI), liver index (LI) and lung index (LuI) and blood eosinophilia during 3 weeks post infectionem (p.i.) with only one challenge. It was found that T. canis larvae arrived at the liver between 10 and 15 hours p.i. by the food-fishhook technique. When the inoculum was more than 200 eggs, there was a reinvasion of liver and lungs at 504 hours p.i. The larval arrival at the brain was delayed in an inverse relationship to the inoculum but the percentage of recovery was about 7 and 8%. The organic indexes were in a direct relationship with the inoculum size and the increase in eosinophilic levels in massive infections (2000 eggs) was in agreement with a strong immune response.

[Frequency of finding helminths eggs in canine stool samples collected in public places in the La Plata city, Argentina]. / Frecuencia de hallazgo de huevos de helmintos en materia fecal canina recolectada en lugares públicos de la ciudad de La Plata, Argentina.

A study was carried out in order to determine the level of contamination with helminth eggs in different sectors of the city of La Plata, Argentina. A total of 351 fecal pools were collected from streets (328) and parks and public places (23) of this locality. Of these fecal pools, 117 (33.3%) contained eggs of at least one helminth species. The overall frequencies found were: Uncinaria sp. 16.0%, Trichuris vulpis 9.7% and Toxocara canis 10.5%.