Cristiane Maria Colli

Association between contamination of public squares and seropositivity for Toxocara spp. in children.

A concomitant study was carried out, of the association of positive serology for Toxocara spp. in 90 children who played in public squares used for leisure, with the frequency with which each child used these areas, and the presence of eggs of Toxocara spp. in the sand or grass in these locations. The sand and grass of their peridomiciles and school playgrounds, as well as the feces of their dogs were also analyzed for Toxocara. Serum samples were tested for IgG antibodies to Toxocara canis excreted-secreted larval antigens by ELISA, and blood samples for eosinophilia. The water-sedimentation technique was used to evaluate the presence of parasite eggs in the sand and grass turfs, and in feces of the dogs that also frequented these locations. 16/90 (17.8%) of the children were seropositive for Toxocara spp. There was a positive association between seropositivity in children who played in the public squares six or seven times a week, with a parasite load above 1.1 eggs/g of sand, as well as with contamination of the peridomicile, even at less than 1.0 egg/g of sand. Eosinophilia, the habit of geophagy, age from one to four years, and the presence of parasitized pet dogs were also positively correlated with seropositivity in the children. Eggs were found in 15/15 (100%) of the public squares, 17/90 (18.9%) of the peridomiciles, 3/13 (23.1%) of the schools, and 12/41 (29.3%) of the dogs living in the peridomiciles investigated.

Serological, clinical and epidemiological evaluation of toxocariasis in urban areas of south Brazil

Toxocariasis is a worldwide public-health problem that poses major risks to children who may accidentally ingest embryonated eggs of Toxocara. The objectives of this study were to investigate the occurrence of anti-Toxocara spp. antibodies in children and adolescents and the variables that may be involved, as well as environmental contamination by Toxocara spp. eggs, in urban recreation areas of north central mesoregion, Paraná State, Brazil. From June 2005 to March 2007. a total of 376 blood samples were collected by the Public Health Service from children and adolescents one to 12 years old, of both genders. Samples were analyzed by the indirect ELISA method for detection of anti-Toxocara antibodies. Serum samples were previously absorbed with Ascaris suum antigens, and considered positive with a reagent reactivity index >1. Soil samples from all of the public squares and schools located in the four evaluated municipalities that had sand surfaces (n = 19) or lawns (n = 15) were analyzed. Of the 376 serum samples, 194 (51.6%) were positive. The seroprevalence rate was substantially higher among children aging one to five years (p = 0.001) and six to eight years (p = 0.022). The clinical signs and symptoms investigated did not show a statistical difference between seropositive and seronegative individuals (p > 0.05). In 76.5% of the investigated recreation places, eggs of Toxocara were detected in at least one of the five collected samples. Recreation areas from public schools were 2.8 times more contaminated than from public squares. It is important to institute educational programs to inform families and educators, as well as to improve sanitary control of animals and cleaning of the areas intended for recreation in order to prevent toxocariasis.

Identical Assemblage of Giardia duodenalis in Humans, Animals and Vegetables in an Urban Area in Southern Brazil Indicates a Relationship among Them

Background Giardia duodenalis infects humans and other mammals by ingestion of cysts in contaminated water or food, or directly in environments with poor hygiene. Eight assemblages, designated A–H, are described for this species. Methodology/Principal Findings We investigated by microscopy or by direct immunofluorescence technique the occurrence of G. duodenalis in 380 humans, 34 animals, 44 samples of water and 11 of vegetables. G. duodenalis cysts present in samples were genotyped through PCR-RFLP of β giardin and glutamate dehydrogenase (gdh) genes and sequencing of gdh. The gdh gene was amplified in 76.5% (26/34) of the human faeces samples with positive microscopy and in 2.9% (1/34) of negative samples. In 70.4% (19/27) of the positive samples were found BIV assemblage. In two samples from dogs with positive microscopy and one negative sample, assemblages BIV, C, and D were found. Cysts of Giardia were not detected in water samples, but three samples used for vegetable irrigation showed total coliforms above the allowed limit, and Escherichia coli was observed in one sample. G. duodenalis BIV was detected in two samples of Lactuca sativa irrigated with this sample of water. BIV was a common genotype, with 100% similarity, between different sources or hosts (humans, animals and vegetables), and the one most often found in humans. Conclusions/Significance This is the first study in Brazil that reports the connection among humans, dogs and vegetables in the transmission dynamics of G. duodenalis in the same geographic area finding identical assemblage. BIV assemblage was the most frequently observed among these different links in the epidemiological chain.

Avaliação eosinofílica e soropositividade para anticorpos IgG anti-toxocara em crianças atendidas pelo Sistema Único de Saúde

Objective: To study the association between the presence of eosinophilia and IgG antibodies to Toxocara spp. in children assisted by the public health service, in the northwestern region of Parana State, in southern Brazil. Methods: A retrospective study of children aged seven months to 12 years old assisted by the Public Health Service in northwest state of Parana, Brazil. ELISA test was performed in all children in order to detect IgG antibodies to Toxocara spp. and eosinophil amounts (eosinophilia ≥ 600 cells/mm3). Results: Among 1,199 screened children, 386 (32.2%) had IgG antibodies to Toxocara spp. The seroprevalence of Toxocara spp. and the eosinophilia were more common among children of seven months to five years old. Eosinophilia was observed in 7.8% of seropositive patients to Toxocara spp. Conclusions: There was a high prevalence of anti-Toxocara spp., mainly in children under five years old. With the Endereco para correspondencia: Ana Lucia F. Guilherme Avenida Colombo, 5.790,Bloco I-90, sala 11 CEP 87020-900 – Maringa/PR E-mail: alfguilherme@uem.br Fonte financiadora: Fundo Parana, o qual apoiou com verba a aquisicao de equipamentos e material de consumo para o projeto. Conflito de interesse: nada a declarar Recebido em: 16/10/2009 Aprovado em: 11/6/2010 Rev Paul Pediatr 2011;29(1):80-4. Artigo Original

Identification of public areas with potential toxocariasis transmission risk using Geographical Information Systems

Geographic information system (GIS) or Health geography is a geographical identification of the spatial variation of contagion disease and zoonosis risk in certain conditions.ObjectiveComparing the variation of environmental contamination of Toxocara spp. sands of lawns and public areas through the Geographic Information System.Materials and MethodsWere investigated 98% of Parana recreational urban areas such as, schools, public parks and squares filled with sand and/or lawn. Samples were analyzed using zinc sulphate solution centrifuge-flotation, density 1.420, and water sedimentation. Maps were developed by ESRI Arc GIS 9.2.ResultsTotal 77/98 (78,6%) of public spaces shown Toxocara spp. eggs presence. No significant difference was observed (p = 0,9999) between eggs prevalence in sand 44/56 (78,6%), and lawn 33/42 (78,6%), at school (p = 0,6898) and squares (p = 0,0616). Egg contamination prevailed in periphery areas in the sand 20/27 (74%) and lawn 18/21 (86%). However, no significant difference was observed between sand and lawn contamination from schools and parks to central and peripheral areas. Although schools 62/66 (93,9%) presented a higher number of enclosed spaces, did not provide a lower frequency of pollution (p = 0,3327).ConclusionGIS observed a scattered pattern of zoonosis among schools and public squares sand and lawns, with predominance of contamination in the peripheral areas with presence of dogs/cats.

Food handlers as a link in the chain of transmission of Giardia duodenalis and other protozoa in public schools in southern Brazil.

BACKGROUND Food handlers (FHs) may facilitate transmission and dissemination of pathogens. The importance of FHs as a link in the epidemiological chain of transmission of Giardia duodenalis and other intestinal protozoa was assessed. METHODS Fecal and subungual material from 27 FHs were analyzed using parasitological methods. G. duodenalis was identified by direct immunofluorescence and genotyped by PCR-RFLP for the bg and gdh genes, and gdh was sequenced. RESULTS At least one protozoan was detected in 30% (8/27) of the FHs and G. duodenalis (19%; 5/27) was the most common species. The AII and BIV genotypes were found in 20% (1/5) and 60% (3/5) of FHs infected with G. duodenalis, respectively. CONCLUSIONS FHs can be involved in the chain of transmission of G. duodenalis and other protozoa. GENBANK ACCESSION NUMBERS KJ741310 - KJ741313.

Prevalence and risk factors for intestinal parasites in food handlers, southern Brazil

In this study, the prevalence and risk factors for enteroparasites were determined in food handlers from Maringá, Paraná State, southern Brazil. Fecal and subungual materials of 150 street food vendors were analyzed by the methods of Lutz, Faust, and Mello, respectively. A questionnaire on hygiene and sanitary conditions of the workplace and of domicile was applied. The prevalence of enteroparasites was 28%, and the protozoa infection was more expressive (21.3%) than by helminths (6.7%), but without significant difference (p > 0.05). Entamoeba coli was the most frequent species occurring in 15.3%, while the prevalence of protozoa pathogenic was low (Giardia lamblia: 2.7% and Entamoeba histolytica: 0.7%). The subungual material presented negative results. The presence of pets in domiciles has increased twice the risk of infection. The working conditions of the majority of street food vendors were inappropriate. The results highlight the need for more rigorous programs of continuing education, parasitological examination every six months, and health surveillance. In this way, the quality of the service provided to the population can be improved and the transmission of food-borne diseases can be prevented.

Simplified protocol for DNA extraction and amplification of 2 molecular markers to detect and type Giardia duodenalis

We evaluated the ability of 3 kits: QIAmp® DNA stool mini kit (Qiagen, Hilden, Germany), PureLink PCR Purification®, and PureLink™ Genomic DNA® (Invitrogen, Carlsbad, CA, USA) for DNA extraction, and of 2 molecular markers (heat shock protein [HSP] and β-giardin genes) for detection and genotyping of Giardia duodenalis stool samples. The detection and typing limits of the markers were determined by the DNA concentration of trophozoites and cysts and were tested in 26 clinical samples. Of the 3 kits tested, the PureLink PCR Purification gave the best results when tested with clinical samples with low, intermediate, and high numbers of cysts. The DNA extracted from trophozoites and cysts was diluted successively in 1:2 ratios until it was no longer possible to observe the amplified product in polyacrylamide gel. Similarly, a suspension of cysts was diluted until no cysts were observed, and then the DNA was extracted. The amount of DNA of trophozoites and cysts for the typing of the parasite was smaller for the HSP marker than for β-giardin. Combined use of both markers allowed us to detect DNA of Giardia in parasitologically positive samples in a higher percentage (75%) than the results obtained for each marker and in 1 parasitologically negative sample, indicating that this combination increased the potential to accurately detect and genotype this parasite. We also concluded that the HSP marker has a higher limit of detection and typing than the β-giardin marker and that the DNA extraction method tested for G. duodenalis is simpler and more efficient than those that are currently in use and can be applied on a large scale.

Predominance of Giardia duodenalis Assemblage AII in Fresh Leafy Vegetables from a Market in Southern Brazil.

We investigated the presence of Giardia duodenalis cysts and its genotypes in raw leafy vegetables sold in a Brazilian market. These products are different from those sold in most street markets because the producers themselves display and sell their products and rely on specialized technical and sanitary assistance. Vegetable and water samples were collected from 14 (80%) producers who cultivated vegetables that are typically consumed raw for sale at the market, obtained at the market and farms, respectively. A total of 128 samples of leafy greens (chives, parsley, cabbage, arugula, watercress, and chicory) and 14 water samples were analyzed by direct immunofluorescence and PCR techniques. The positive samples were genotyped (GHD gene) using PCR and restriction fragment length polymorphism. The analyses indicated that 16 (12.5%) of 128 samples were positive by PCR, while 1 (0.8%) of 128 samples were positive by immunofluorescence. Giardia cysts were not detected in the water samples obtained at the farms. The molecular technique revealed a genotype with zoonotic potential, which underscores the challenge in the control of giardiasis dissemination via the consumption of raw vegetables.

Eosinophilic count and seropositivity for IgG antibodies to toxocara spp. in children assisted at the public health service

Objective: To study the association between the presence of eosinophilia and IgG antibodies to Toxocara spp. in children assisted by the public health service, in the northwestern region of Parana State, in southern Brazil. Methods: A retrospective study of children aged seven months to 12 years old assisted by the Public Health Service in northwest state of Parana, Brazil. ELISA test was performed in all children in order to detect IgG antibodies to Toxocara spp. and eosinophil amounts (eosinophilia ≥ 600 cells/mm3). Results: Among 1,199 screened children, 386 (32.2%) had IgG antibodies to Toxocara spp. The seroprevalence of Toxocara spp. and the eosinophilia were more common among children of seven months to five years old. Eosinophilia was observed in 7.8% of seropositive patients to Toxocara spp. Conclusions: There was a high prevalence of anti-Toxocara spp., mainly in children under five years old. With the Endereco para correspondencia: Ana Lucia F. Guilherme Avenida Colombo, 5.790,Bloco I-90, sala 11 CEP 87020-900 – Maringa/PR E-mail: alfguilherme@uem.br Fonte financiadora: Fundo Parana, o qual apoiou com verba a aquisicao de equipamentos e material de consumo para o projeto. Conflito de interesse: nada a declarar Recebido em: 16/10/2009 Aprovado em: 11/6/2010 Rev Paul Pediatr 2011;29(1):80-4. Artigo Original