Genotyping of Giardia duodenalis in vegetables cultivated with organic and chemical fertilizer from street markets and community vegetable gardens in a region of Southern Brazil.

Background: In order to investigate the occurrence of Giardia duodenalis and its genotypes in vegetables that are consumed raw, we analyzed samples cultivated with organic or chemical fertilizer, sold in street markets and from community vegetable gardens in an urban area located in Southern Brazil. Methods: We analyzed 130 samples of vegetables such as crisp lettuce, regular lettuce, kale, chicory and rocket, from street markets, and 130 from community gardens. From each sample, 50 g were washed in Tween 80 solution (1%) and the solution obtained was filtered through a cellulose acetate membrane. The retained material was used for DNA extraction with the commercial kit Purelink®. GDH gene was amplified by semi-nested PCR using the GDHeF, GDHiR and GDHiF primers. Positive samples were genotyped using the PCR-RFLP technique with the restriction enzyme NlaIV. Results: We obtained 7.3% (19/260) positive samples for G. duodenalis, both from street markets (10/130) and from community gardens (9/130), including organic and non-organic products. The assemblage AI was predominant, but assemblages B and E were also found. Conclusions: The molecular technique revealed genotypes with zoonotic potential, evidencing the importance of investigating commercialized vegetables that are consumed raw and establishing a more rigid quality control.

First Detection of Toxoplasma gondii DNA in the Fresh Leafs of Vegetables in South America.

Despite the high dispersion of Toxoplasma gondii oocysts in the environment, there are few studies investigating their presence in vegetables consumed by the general population. This has led us to investigate its occurrence in raw vegetables. A total of 238 samples of vegetables were collected, including crisp lettuce, regular lettuce, chicory, rocket, and parsley, both organic and nonorganic, locally in northwestern Parana, Southern Brazil. Each sample (50 g) was washed and filtered separately. A PCR was performed to detect the parasite DNA from the sediment of each sample, using B1 (B22-23) and Toxo4-5 primers. We found contamination in 3.8% of the samples, 0.8% with the primer Toxo4-5 and 2.9% with B22-B23. The results were positive in 0.6% (1/62) of the samples of smooth lettuce, 3.7% (4/106) of crisp head lettuce, 5.0% (2/40) of chicory, 14.3% (1/7) of rocket, and 20% (1/5) of parsley. These data show the contamination by T. gondii in raw vegetables directly from production sites and stores, in both organic and nonorganic samples.

PATHOGENICITY OF Blastocystis sp. TO THE GASTROINTESTINAL TRACT OF MICE: RELATIONSHIP BETWEEN INOCULUM SIZE AND PERIOD OF INFECTION / Histopatogenicidade de Blastocystis sp. para o trato gastrointestinal de camundongos: relação com inóculo e tempo de infecção

The pathogenic potential of Blastocystis sp. in experimental models requires further investigation. In this work, the pathogenicity of this parasite in the gastrointestinal tract of male Swiss mice was evaluated according to the inoculum size and period of infection. Animals were infected intragastrically, with 100, 500, 1,000, 5,000 and 10,000 Blastocystis sp. vacuolar forms obtained from a mixture of eight human isolates cultured axenically in Jones' medium. After seven, 14, 21, 28 and 60 days of infection, the animals were sacrificed and fragments of the small intestine (duodenum), large intestine, and cecum were subjected to histopathological analysis. Blastocystis sp. triggered an inflammatory response in the different tissues analyzed, with a predominance of mononuclear cells. The parasite was found in the muscular layer of the cecum, showing its invasive character. Larger inocula triggered inflammatory processes earlier (seven days) than smaller ones (from 21 days). We conclude that, in the proposed model, the pathogenicity of Blastocystis sp. isolates that were studied is related to inoculum size and period of infection.

Pouco é sabido sobre o potencial patogênico de Blastocystis sp. em modelos experimentais. Neste trabalho a patogenicidade desse parasito para o trato gastrointestinal de camundongos Swiss machos foi avaliada de acordo com o inóculo e tempo de infecção. Os animais foram infectados, via intragástrica, com 100, 500, 1.000, 5.000 e 10.000 formas vacuolares de Blastocystis sp. obtidos a partir de uma mistura de oito isolados humanos cultivados axenicamente em meio Jones. Após 7, 14, 21, 28 e 60 dias de infecção os animais foram sacrificados e fragmentos do intestino delgado (duodeno), grosso e ceco foram retirados para análise histopatológica. Blastocystis sp. desencadeou resposta inflamatória nos diferentes tecidos analisados, com predominância de infiltrado mononuclear. No ceco o parasito foi encontrado na túnica muscular mostrando seu caráter invasivo. Inóculos maiores desencadearam processos inflamatórios mais precocemente (7 dias) e inóculos menores mais tardiamente (a partir de 21 dias). Conclui-se que no modelo proposto a patogenicidade dos isolados de Blastocystis sp. estudados tem relação com o inóculo e tempo de infecção.

PATHOGENICITY OF Blastocystis sp. TO THE GASTROINTESTINAL TRACT OF MICE: RELATIONSHIP BETWEEN INOCULUM SIZE AND PERIOD OF INFECTION.

The pathogenic potential of Blastocystis sp. in experimental models requires further investigation. In this work, the pathogenicity of this parasite in the gastrointestinal tract of male Swiss mice was evaluated according to the inoculum size and period of infection. Animals were infected intragastrically, with 100, 500, 1,000, 5,000 and 10,000 Blastocystis sp. vacuolar forms obtained from a mixture of eight human isolates cultured axenically in Jones' medium. After seven, 14, 21, 28 and 60 days of infection, the animals were sacrificed and fragments of the small intestine (duodenum), large intestine, and cecum were subjected to histopathological analysis. Blastocystis sp. triggered an inflammatory response in the different tissues analyzed, with a predominance of mononuclear cells. The parasite was found in the muscular layer of the cecum, showing its invasive character. Larger inocula triggered inflammatory processes earlier (seven days) than smaller ones (from 21 days). We conclude that, in the proposed model, the pathogenicity of Blastocystis sp. isolates that were studied is related to inoculum size and period of infection.

Identification of public areas with potential toxocariasis transmission risk using geographical information systems.

UNLABELLED: Geographic information system (GIS) or Health geography is a geographical identification of the spatial variation of contagion disease and zoonosis risk in certain conditions. OBJECTIVE: Comparing the variation of environmental contamination of Toxocara spp. sands of lawns and public areas through the Geographic Information System. MATERIALS AND METHODS: Were 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. RESULTS: Total 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). CONCLUSION: GIS 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.

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.

Serological, clinical and epidemiological evaluation of toxocariasis in urban areas of south Brazil / Avaliação sorológica, clínica e epidemiológica da toxocaríase em áreas urbanas do sul do Brasil

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 percent) 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 percent 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.

A toxocaríase é um problema de saúde pública mundial, com maior risco para crianças que podem, acidentalmente, ingerir ovos embrionados de Toxocara spp.. Os objetivos deste estudo foram avaliar a ocorrência de anticorpos anti-Toxocara spp. em crianças e adolecentes e as variáveis que podem estar envolvidas, bem como a contaminação ambiental por ovos de Toxocara spp., em locais de recreação, em áreas urbanas da mesorregião norte central, Paraná, Brasil. De junho de 2005 a março de 2007 foram coletadas 376 amostras de sangue de crianças e adolescentes de um a doze anos, de ambos os sexos, atendidas pelo Sistema Único de Saúde. As amostras foram analisadas pelo método de ELISA indireto para detecção de IgG anti-Toxocara e previamente absorvidas com antígeno de Ascaris suum. Foram consideradas reagentes as amostras com índice de reatividade > 1. A análise das amostras de areias (n = 19) e gramados (n = 15) de cada município foi realizada em todas as praças e escolas públicas. Das 376 amostras de soro, 194 (51,6 por cento) foram positivas. A taxa de soroprevalência foi substancialmente mais elevada entre as crianças na faixa etária de até um a cinco (p = 0.001) e de seis a oito anos de idade (p = 0,022). Os sinais e sintomas clínicos investigados não mostraram diferenças estatísticas entre soropositivos e soronegativos (p > 0,05). Em 76,5 por cento dos locais de recreação investigados, ovos de Toxocara foram detectados em pelo menos uma das cinco amostras. Os locais de recreação das escolas públicas estavam 2,8 vezes mais contaminados do que as praças. É importante a realização de programas educativos junto às famílias e educadores, o controle sanitário de animais e a higienização dos locais destinados à recreação para prevenção da toxocaríase.