Larva migrans cutânea: ocorrência de casos humanos e identificação de larvas de Ancylostoma spp em parque público do município de Taciba, São Paulo

Descreve-se a ocorrência de larva migrans cutânea em crianças de Taciba, São Paulo, Brasil, pelo contato com areia de parques públicos, onde observou-se larvas de Ancylostoma spp. A enfermidade foi controlada após adotar-se medidas educacionais sanitárias, desativação e isolamento dos locais para reduzir acesso de animais e troca de areia.

Cross-reactions between Toxocara canis and Ascaris suum in the diagnosis of visceral larva migrans by western blotting technique

Visceral larva migrans (VLM) is a clinical syndrome caused by infection of man by Toxocara spp, the common roundworm of dogs and cats. Tissue migration of larval stages causes illness specially in children. Because larvae are difficult to detect in tissues, diagnosis is mostly based on serology. After the introduction of the enzyme-linked immunosorbent assay (ELISA) using the larval excretory-secretory antigen of T. canis (TES), the diagnosis specificity was greatly improved although cross-reactivity with other helminths are still being reported. In Brazil, diagnosis is routinely made after absorption of serum samples with Ascaris suum antigens, a nematode antigenicaly related with Ascaris lumbricoides which is a common intestinal nematode of children. In order to identify T. canis antigens that cross react to A. suum antigens we analyzed TES antigen by SDS-PAGE and Western blotting techniques. When we used serum samples from patients suspected of VLM and positive result by ELISA as well as a reference serum sample numerous bands were seen (molecular weight of 210-200 kDa, 116-97 kDa, 55-50 kDa and 35-29 kDa). Among these there is at least one band with molecular weight around 55-66 kDa that seem to be responsible for the cross-reactivity between T. canis and A. suum once it disappears when previous absorption of serum samples with A. suum antigens is performed.

Application of a species-specific PCR-RFLP to identify Ancylostoma eggs directly from canine faeces

Apart from their veterinary importance, the hookworms Ancylostoma caninum, Ancylostoma braziliense and Ancylostoma caninum are also capable of causing zoonotic disease in humans. A highly sensitive and species-specific PCR-RFLP technique was utilised to detect and differentiate the various canine Ancylostoma spp directly from eggs in faeces. This technique was utilised to screen 101 canine faecal samples from parasite endemic tea growing communities in Assam, India, as part as an ongoing epidemiological investigation into canine parasitic zoonoses. The prevalence of hookworms in dogs was found to be 98% using a combination of PCR and conventional microscopy. Overall, 36% of dogs were found positive for single hookworm infections with A. caninum, 24% positive for single infections with A. braziliense and 38% had mixed infections with both A. caninum and A. braziliense. No dogs were found positive for A. ceylanicum in the community under study. The high prevalence of A. caninum and A. braziliense in dogs in this community may account for the high incidence of cutaneous larva migrans (CLM) observed among the human population residing at the tea estates. The PCR-RFLP technique described herein allows epidemiological screening of canine hookworms to be conducted rapidly, with ease and accuracy, and has the potential to be applied to a number of different clinical, pharmacological and epidemiological situations. (C) 2004 Elsevier B.V. All rights reserved.

Canine gastrointestinal parasitic zoonoses in India

Although well recognized and studied in developed countries, canine parasitic zoonoses pose a lowly prioritized public health problem in developing countries such as India, where conditions are conducive for transmission. A study of the most recent parasite survey determining prevalence and epidemiology of canine parasitic zoonoses among tea-growing communities of northeast India demonstrated the endemicity of the problem. This particular study serves as a model using conventional, as well as molecular parasitological, tools to provide novel insights into the role of dogs as mechanical transmitters of human parasites such as Ascaris and Trichuris, and discusses the risks dogs pose with regards to zoonotic transmission of hookworms and Giardia.

Occurrence of Ancylostoma in dogs, cats and public places from Andradina city, São Paulo state, Brazil

The aim of this study was to determine the frequency and intensity of Ancylostoma spp. in 33 dogs and 52 cats by means of coproparasitological examinations and parasitological necropsy, and assess the presence of contaminated feces with eggs of that parasite in public places of Andradina Municipality, São Paulo State, Brazil. Willis-Mollay and Sedimentation methods indicated Ancylostoma spp. eggs in 87.8% (29/33) dogs and 94.2% (49/52) cats. The species A. caninum and A. braziliense were found in 63.6% (21/33) and 30.3% (10/33) of dogs, respectively. Considering cats, 67.3% (35/52) were parasitized by A. braziliense, 21.1% (11/52) by A. caninum, and 9.6% (5/52) by A. tubaeforme. Forty-two canine fecal samples were collected from public environments, including 23 squares/gardens and 19 streets/sidewalks. Positive samples for Ancylostoma spp. accounted for 64.3% (27/42); squares/gardens had 60.9% (14/23) positive samples, and streets and sidewalks, 68.4% (13/19). No association was observed between the number of Ancylostoma spp parasites and age, sex and breed of the animals and also the ratio of EPG counts and the parasitic intensity observed at necropsy (p > 0.05). Based on the high occurrence of hookworm in dogs and cats in this study, the treatment with anti helminthics are needed even in those animals with negative stool tests, besides adopting control of the number of animals in public places, in order to decrease the likelihood of environmental contamination, since this parasite represents a potential hazard to human and animal health.

Contamination of public parks and squares from Guarulhos (São Paulo State, Brazil ) by Toxocara spp. and Ancylostoma spp.

The contaminated soil with mammal feces is an important factor of risk to infection with zoonotic diseases. Amongst these zoonoses are visceral larva migrans and cutaneous larva migrans caused by Toxocara spp. and Ancylostoma spp., respectively. The aim of this study was to assess the environmental contamination by Toxocara spp. eggs and hookworms (Ancylostoma spp.) in public parks and squares in the city of Guarulhos, a metropolitan area of São Paulo, São Paulo State, Brazil. Soil samples were collected, between September and December 2010, and examined using the centrifugal flotation technique with sodium dichromate and zinc sulphate as well as the modified Baermann method. Notably, 35 (74.5%) of the 47 districts surveyed in Guarulhos possessed samples contaminated with Toxocara spp. and/or eggs or larvae of Ancylostoma spp. The frequency of Toxocara spp. and Ancylostoma spp. in the samples from public areas was 68.1% and 46.8%, respectively. Overall, the eastern side of Guarulhos is the region with the highest occurrence of causative agents of larva migrans. In all collection sites, the presence of feces from dogs and cats accompanied by their owners and stray animals were observed. Notably, it is important to adopt measures to control dog and cat breeding, to treat infected animals, and provide health education to the population.

Occurrence of Ancylostoma in dogs, cats and public places from Andradina city, São Paulo state, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Sero-epidemiology of toxocariasis in a rural settlement in Sao Paulo state, Brazil

The seroprevalence of Toxocara canis and risk factors for infection with this parasite were explored in a rural settlement in Sao Paulo state, Brazil. Total IgA and IgE levels in 79 subjects were determined by turbidimetry and chemiluminescence, respectively. Total counts of leucocytes and erythrocytes and differential counts of leucocytes were made by flow cytometry. ELISA for the detection of anti-Toxocara IgG, IgA and IgE were standardized using Toxocara excretory-secretory antigens (TES) obtained from the cultured second-stage larvae of T. canis. Seventeen (21.5%) of the subjects were found positive for anti-Toxocara IgG, with no significant differences in such seropositivity with age or gender. Thirty (38%) of the subjects showed eosinophilia and 70 (89%) had elevated levels of total IgE. Among the 17 subjects found seropositive for anti-Toxocara IgG, the percentage of leucocytes represented by eosinophils (P=0.0069) and total levels of IgE (P=0.0452) were positively correlated with the levels of anti-TES IgE. Although anti-TES IgA was detected in 10 (59%) of the subjects, there was no significant correlation between the levels of total IgA and those of Toxocara-specific IgA. Only one of the 17 subjects found positive for anti-Toxocara IgG had attended a secondary school and all but two belonged to households with monthly incomes of < U.S.$100. In the study community at least, seropositivity may be related to poor living standards and lack of basic sanitary conditions. The presence of anti-Toxocara IgE and IgA may facilitate the diagnosis of toxocariasis and may well be useful for monitoring the success of treatment.

Ovos de Toxocara sp. e larvas de Ancylostoma sp. em praça pública de Lavras, MG

Larva migrans visceral e cutânea são zoonoses parasitárias causadas pela infecção da larva de Toxocara sp. e Ancylostoma sp., respectivamente. O objetivo do estudo foi verificar a contaminação por ovos de Toxocara sp. e ovos e larvas de Ancylostoma sp. em amostras de solos coletadas de praças públicas e de áreas de recreação infantil de Lavras, Estado de Minas Gerais, por meio da técnica de centrífugo-flutuação e do método de Baermann. A ocorrência de ovos de Toxocara sp. e, ovos e larvas de Ancylostoma sp. foi observada em 69,6% (16/23) das amostras de solo coletadas de praças públicas. A contaminação somente por ovos de Ancylostoma sp. em amostras de solo coletadas em escolas/creches foi de 22,2% (4/18). A percentagem de amostras de areia coletadas de escolas/creches contaminadas somente com larvas de Ancylostoma sp. foi de 11,1% (2/18). Praças públicas são as áreas com maior risco potencial de infecção por Toxocara sp. e Ancylostoma sp. Exame coproparasitológico realizado em 174 amostras de fezes de cães observou 58% e 23%, respectivamente, com ovos de Ancylostoma sp. e Toxocara sp.

Occupational and domiciliary contact with dogs as factors of risk to human infection with Toxocara larvae

The contact with dogs at home or place of work has been investigated as factors of risk in the occurrence of the visceral larva migrans syndrome caused by Toxocara, in man. Through the E.L.I.S.A. (enzyme-linked immunosorbent assay) technique, the presence of antibodies to Toxocara was searched in the sera of 79 women who have been raising or had raised dogs at home in the last two years and 123 men, who were municipal public employees in charge of the capture and keeping of stray dogs. The control groups were constituted by 205 sera from women who denied domiciliary contact with dogs, at least in the last two years, and 139 sera from men whose occupation did not urge them to contact with dogs. A significant more elevated frequency of antibodies to Toxocara was observed among women with domiciliary contact with dogs; nevertheless, there was not a significant difference in the positive rates in the case of men with occupational contact with dogs.

Standardization of dot-ELISA for the serological diagnosis of toxocariasis and comparision of the assay with ELISA

The dot-enzyme-linked immunosorbent assay (dot-ELISA) was standardized using somatic (S) and excretory-secretory (ES) antigens of Toxocara-canis for the detection of specific antibodies in 22 serum samples from children aged 1 to 15 years, with clinical signs of toxocariasis. Fourteen serum samples from apparently normal individuals and 28 sera from patients with other pathologies were used as controls. All samples were used before and after absorption with Ascaris suum extract. When the results were evaluated in comparison with ELISA, the two tests were found to have similar sensitivity, but dot-ELISA was found to be more specific in the presence of the two antigens studied. Dot-ELISA proved to be effective for the diagnosis of human toxocariasis, presenting advantages in terms of yield, stability, time and ease of execution and low cost.

Clinical and laboratorial features of visceral toxocariasis in infancy

Forty children with a diagnosis of Visceral Toxocariasis were evaluated prospectively from February 1982 to June 1989. Diagnosis was established by clinical, laboratorial and serological (ELISA - ES Toxocara canis antigen) evaluations. A great clinical polymorphism was found in our patients, ranging from unspecific or absent manifestations to an exhuberant symptomatology. The laboratorial findings were: leukocytosis,eosinophilia and elevation of serum gammaglobulin and isohemagglutinin levels. No significant relationship between clinical findings and laboratorial parameters was found. Serology (ELISA) was a method of great diagnostic support but did not show a correlation with clinical and laboratorial findings in this study. There was a significant relationship between pulmonary manifestations and the presence of signs and/or symptoms, when the patients were sent to us. Our findings, especially the high incidence of pulmonary manifestations, suggest that Visceral Toxocariasis has to be included in the differential diagnostic of children with pulmonary manifestations, characteristic epidemiological data and associated eosinophilia.

PRESENCE OF ANTI-TOXOCARA ANTIBODIES IN CHILDREN SELECTED AT HOSPITAL UNIVERSITÁRIO, CAMPO GRANDE, MS, BRAZIL

Visceral Larva Migrans syndrome (VLM) results from the presence or migration of helminth larvae in humans, who nonetheless only play the role of paratenic hosts in the helminths' life cycle. In humans, VLM can be caused by larvae of various nematode species, chiefly those of the ascarid Toxocara canis, which can then be found at a variety of body sites, such as the liver, lungs, heart, and brain. Clinical and pathological manifestations depend primarily on larvae number and location, infection duration, reinfection occurrence, and host's immunological condition. Signs and symptoms may range from asymptomatic infection to severe disease. In humans, infection is acquired through ingestion of T. canis eggs present in soil, containing larvae in the infective stage7, 8, 9. Indeed, eggs of Toxocara sp. have been found in sandboxes in several public places in the city of Campo Grande, Mato Grosso do Sul state2. This study was carried out to detect the presence of anti-Toxocara antibodies in children attending the Pediatrics division of Hospital Universitário of Universidade Federal de Mato Grosso do Sul at Campo Grande, Brazil. Over the years 1992-94, 454 serum samples, obtained from children of 5.25 ± 3.28 years of mean age and selected at that hospital on the basis of eosinophil count greater than or equal to 1000/mm3 of blood, were tested for the presence of antibodies by means of the ELISA technique employing Toxocara canis larvae excretory-secretory antigens5. A high prevalence rate for toxocariasis (35.55%) was found, which was observed to be associated with eosinophil levels lower than those usually reported in literature. Furthermore, a higher frequency of positive serology among boys was also observed (13 cases in contrast to only 3 among girls), a result also reported by other authors

PREVALENCE OF ANTI- Toxocara ANTIBODIES IN A RANDOM SAMPLE OF INPATIENTS AT A CHILDREN'S HOSPITAL IN VITÓRIA, ESPÍRITO SANTO, BRAZIL

In the streets of Vitória, in the State of Espírito Santo, Brazil, are large number of stray dogs, many of which are infected with Toxocara canis, suggesting a high risk for human infection. In order to investigate the prevalence of Toxocara infection in children in Espírito Santo we studied the prevalence of anti-Toxocara antibodies in 100 random inpatients over one year of age, at the Children's Hospital N.S. da Glória, the reference children's hospital for the State.All the sera were collected during the period between October 1996 and January 1997. The mean age was 6.6±4.1 yrs. (1 to 14 yrs., median 6yrs.) and there were patients from all of the different wards of the hospital. Sixty-eigth patients came from the metropolitan area of Vitória and the other 32 from 17 other municipalities. The anti-Toxocara antibodies were investigated by ELISA-IgG using a secretory-excretory antigen obtained from second stage larvae. All sera were adsorbed with Ascaris suum antigen before the test. Thirty-nine sera (39%) were positive, predominantly from boys, but the gender difference was not statistically significant (boys:25/56 or 44.6%; girls:14/44 or 31.8%; p=0.311). The prevalence of positive sera was higher, but not statistically significant, in children from the urban periphery of metropolitan Vitória (formed by the cities of Vitória, Cariacica, Vila Velha, Serra and Viana) than in children from 17 other municipalities (44.1% and 28.1% respectively, p=0.190). Although the samples studied do not represent all children living in the State of Espírito Santo, since the Children's Hospital N.S. da Glória admits only patients from the state health system, it is probable that these results indicate a high frequency of Toxocara infection in children living in Espírito Santo. Further studies of population samples are necessary to ascertain the prevalence of Toxocara infection in our country.

TOXOCARIASIS: SEROLOGICAL DIAGNOSIS BY INDIRECT ANTIBODY COMPETITION ELISA

Toxocariasis is caused by infection of man by Toxocara canis and Toxocara cati larvae, the common roundworm of dogs and cats. Because larvae are difficult to detect in tissues, diagnosis is mostly based on serology. Non specific reactions are observed mainly due to cross-reactivity with Ascaris sp antigens. This investigation aimed at developing and evaluating an indirect antibody competition ELISA (IACE) employing a specific rabbit IgG anti-Toxocara canis excretory-secretory antigens as the competition antibody, in order to improve indirect ELISA specificity performed for toxocariasis diagnosis. For that, the rabbit IgG was previously absorbed by Ascaris suum adult antigens. Sensitivity and specificity of IACE were first evaluated in 28 serum samples of mice experimentally infected with T. canis embryonated eggs. Adopting cut-off value established in this population before infection, sensitivity and specificity were 100% after 20 days post-inoculation. For human population IACE was evaluated using sera from 440 patients with clinical signs of toxocariasis and the cut-off value was established with 60 serum samples from apparently healthy individuals. Using as reference test the indirect ELISA performed by Adolfo Lutz Institute, sensitivity was 60.2%, specificity was 98% and concordance was 77.3%. Repeatability of IACE was evaluated by the inter-reactions variation coefficient (2.4%).