ANTHELMINTIC ACTIVITY OF LAPACHOL, β-LAPACHONE AND ITS DERIVATIVES AGAINST Toxocara canis LARVAE

Anthelmintics used for intestinal helminthiasis treatment are generally effective; however, their effectiveness in tissue parasitosis (i.e. visceral toxocariasis) is moderate. The aim of this study was to evaluate the in vitroactivity of lapachol, β-lapachone and phenazines in relation to the viability of Toxocara canis larvae. A concentration of 2 mg/mL (in duplicate) of the compounds was tested using microculture plates containing Toxocara canis larvae in an RPMI-1640 environment, incubated at 37 °C in 5% CO2 tension for 48 hours. In the 2 mg/mL concentration, four phenazines, lapachol and three of its derivatives presented a larvicide/larvistatic activity of 100%. Then, the minimum larvicide/larvistatic concentration (MLC) test was conducted. The compounds that presented the best results were nor-lapachol (MLC, 1 mg/mL), lapachol (MLC 0.5 mg/mL), β-lapachone, and β-C-allyl-lawsone (MLC, 0.25 mg/mL). The larvae exposed to the compounds, at best MLC with 100% in vitro activity larvicide, were inoculated into healthy BALB/c mice and were not capable of causing infection, confirming the larvicide potential in vitro of these compounds.

Comparação antigênica entre extratos de Toxocara canis e Ascaris suum

O autor apresenta a comparação de dois extratos de Toxocara canis e um de Ascaris suum do ponto de vista antigênico. O método usado foi a imunodifusão em gel segundo Ouchtehlöny. O extrato de T. canis obtido pela técnica de Jeska mostrou 7 faixas de precipitação contra o anti-soro homólogo. O extrato de A. suum obtido pela mesma técnica apenas revelou 3 faixas, uma das quais com identidade parcial com o antígeno anterior. O extrato de T. canis segundo Woodruff e Thacker revelou 3 faixas de precipitação contra o anti-soro homólogo, todas elas com identidade total com o outro extrato do mesmo parasita.

Environmental contamination by Toxocara sp eggs in public areas of Salvador, Bahia state, Brazil

A survey ofparasite eggs and cysts in soil and dog feces collected in public places of 23 boroughs of Salvador, a city in the Northeast of Brazil, was performed. High degree of contamination by Toxocara sp eggs was observed in all boroughs studied; other parasites found included: Ascaris lumbricoides, hookworms, whipworms and protozoan cysts. Parks andpublic gardens were more contaminated than streets and beachesfor all parasites, including Toxocara sp.

Freqüência de soropositividade para antígenos de Toxocara canis em crianças de classes sociais diferentes

Para estudar a freqüência da infecção pelo Toxocara canis em crianças de classes sociais diferentes de Brasília, Brasil, foram testados soros de 602 crianças de ambos os sexos, de 1 a 12 anos, distribuídas em dois grupos representativos de condições socioeconômicas distintas. As amostras do primeiro grupo foram obtidas em laboratório público que atende bairros pobres. As do segundo grupo foram colhidas em laboratório privado, que serve à classe média. Os anticorpos anti-Toxocara foram detectados pelo método ELISA, com antígenos de Toxocara canis, e absorção com antígenos do Ascaris suum. A prevalência de soropositividade foi de 21,8% (66/302) no primeiro grupo e de 3% (9/300) no segundo (p<0,0001). Não houve diferença quanto ao gênero ou idade das crianças. Os resultados sugerem significativa prevalência de toxocaríase na população infantil de Brasília, com predomínio nas classes sociais menos favorecidas economicamente.

Freqüência de anticorpo anti-Toxocara canis em comunidade do Rio Uatumã, no Estado do Amazonas

Um estudo seccional foi realizado nas Vilas Waimiri e Atroari em Balbina, entre julho e outubro de 2006, com o objetivo de estimar a freqüência de anticorpo antiToxocara canis da classe IgG e avaliar as variáveis epidemiológicas e socioculturais. Foram estudadas 34 famílias e incluídos 100 indivíduos, o que correspondeu a 5% (100/2.000) da população das vilas. A idade variou de zero a 76 anos (M=22,9 Dp=18). Quanto ao gênero, 53% eram femininos e 47% masculino; 52% das amostras foram positivas para Toxocara canis, 44,5% negativas e 3,2% inconclusivas. Observou-se menor número de indivíduos com sorologia negativa na Vila Atroari 29,5% (13/44) em comparação com a Waimiri 46,4% (26/56). Com relação ao contato com cães, dos 55 indivíduos com contato domiciliar 60% (33/55) foram positivos para anticorpo antiToxocara canis Apresentaram sorologia positiva 66,6% (10/15) dos indivíduos que tinham contato domiciliar com filhotes de cão (chi²22,149 p=0,008). A existência de contato domiciliar com cães e filhotes mostrou associação com a presença de anticorpo anti-Toxocara canis na população estudada.

Destruição de ovos de Toxocara canis pelo fungo nematófago Pochonia chlamydosporia

INTRODUÇÃO: Toxocara canis é um ascarídeo parasita do intestino delgado de cães, causador da larva migrans visceral em seres humanos. MÉTODOS: Com o objetivo de demonstrar a eficácia do fungo Pochonia chlamydosporia sobre ovos de Toxocara canis em condições laboratoriais, foi montado ensaio experimental em placas de Petri com ágar-água 2%. RESULTADOS: Houve atividade ovicida de 43,8% (p<0,01) do grupo tratado em relação ao grupo controle durante os intervalos estudados. CONCLUSÕES: Os resultados demonstrados no presente trabalho sugerem a empregabilidade de Pochonia chlamydosporia como uma alternativa de controle biológico dos ovos embrionados de Toxocara canis.

Prevalência e fatores de risco da infecção humana por Toxocara canis em Salvador, Estado da Bahia

INTRODUÇÃO: Larva migrans visceral é causada por Toxocara sp e nunca foi estudada na Bahia. Neste trabalho, investigou-se a prevalência e fatores de risco de infecção por Toxocara canis, em indivíduos de Salvador. MÉTODOS: Trezentos e trinta e oito indivíduos foram investigados para presença de anticorpos IgG séricos anti-T. canis. RESULTADOS: IgG anti-T. canis foi mais alta em indivíduos de classe social baixa com maior contato com cães e gatos, indicando que estas variáveis são fatores de risco para esta infecção. CONCLUSÕES: A prevalência de infecção por T. canis foi alta. Os fatores de risco desta infecção encontrados estão de acordo com a literatura.

Infección por Toxocara Canis: detección en suelo y en niños y adolescentes de comunidades de la provincia de Córdoba

Toxocara canis es un parásito cosmopolita frecuentemente hallado en el intestino delgado de los caninos. El hombre adquiere la infección con Toxocara por la ingestión de huevos embrionados que se encuentran en el suelo contaminado. En Argentina, las cifras reales de prevalencia de esta infección no se conocen por tratarse de una patología que no es de notificación obligatoria y por la existencia de casos asintomáticos. Los objetivos de este proyecto de investigación son: a) determinar el grado de contaminación con huevos de Toxocara canis de suelos en áreas de uso público y privado de las comunas de Villa El Prado y Los Cedros de la Provincia de Córdoba, b) detectar la presencia de anticuerpos anti-Toxocara canis en niños y adolescentes de estas Comunas, c) relacionar la presencia de anticuerpos con factores de riesgo y d) obtener antígenos excresión/secreción de larvas L2 de Toxocara canis. La presencia de huevos de Toxocara se evaluará en suelo (tierra, arena, mixta) del área de recreo de las escuelas y de áreas externas de cada vivienda (patio, jardín). La presencia de anticuerpos específicos será detectada en niños y adolescente entre 1 a 15 años de edad de ambos sexos que asistan a los dispensarios de las comunas. Para elaborar un sistema de detección de anticuerpos específicos se obtendrán antígenos excresión/secreción de larvas L2 de Toxocara canis. Este proyecto permitirá comparar la seroprevalencia de infección por Toxocara en niños y adolescentes, con otras regiones del país y el extranjero y analizar la relación entre títulos y manifestaciones clínicas. Además, posibilitará caracterizar asociaciones entre infección y factores de riesgo. Debido a la carencia de información sobre esta infección parasitaria en nuestro medio, esta investigación aportará datos útiles para las campañas de desparasitación de mascotas, saneamiento ambiental, tratamiento antiparasitario en personas afectadas y caracterización de manifestaciones clínicas asociadas a esta parasitosis.

Infección por toxocara canis: detección en suelo y en niños y adolescentes de comunidades de la provincia de Córdoba (Programa: enfermedades transmisibles y emergentes).

Toxocara canis es un parásito cosmopolita frecuentemente hallado en el intestino delgado de los caninos. El hombre adquiere la infección con Toxocara por la ingestión de huevos embrionados que se encuentran en el suelo contaminado. En Argentina, las cifras reales de prevalencia de esta infección no se conocen por tratarse de una patología que no es de notificación obligatoria y por la existencia de casos asintomáticos. Los objetivos de este proyecto de investigación son: a) determinar el grado de contaminación con huevos de Toxocara canis de suelos en áreas de uso público y privado de las comunas de Villa El Prado y Los Cedros de la Provincia de Córdoba, b) detectar la presencia de anticuerpos anti-Toxocara canis en niños y adolescentes de estas Comunas, c) relacionar la presencia de anticuerpos con factores de riesgo y d) obtener antígenos excreción/secreción de larvas L2 de Toxocara canis para el desarrollo de un técnica inmunoenzimática "in house". La presencia de huevos de Toxocara se evaluará en suelo (tierra, arena, mixta) del área de recreo de las escuelas y de áreas externas de cada vivienda (patio, jardín). La presencia de anticuerpos específicos será detectada en niños y adolescente entre 1 a 15 años de edad de ambos sexos que asistan a los dispensarios de las comunas. Para elaborar un sistema de detección de anticuerpos específicos se obtendrán antígenos excreción/secreción de larvas L2 de Toxocara canis. Este proyecto permitirá comparar la seroprevalencia de infección por Toxocara en niños y adolescentes, con otras regiones del país y el extranjero y analizar la relación entre títulos y manifestaciones clínicas. Además, posibilitará caracterizar asociaciones entre infección y factores de riesgo. Debido a la carencia de información sobre esta infección parasitaria en nuestro medio, esta investigación aportará datos útiles para las campañas de desparasitación de mascotas, saneamiento ambiental, tratamiento antiparasitario en personas afectadas y caracterización de manifestaciones clínicas asociadas a esta parasitosis.

Human and dogs Toxocara canis infection in a poor neighborhood in Bogota

Prevalence of Toxocara canis antibodies was studied in a poor community of Bogotá, Colombia. Two-hundred-sevem patients, from both sexes and all age groups, were studied. Positive Elisa titers were found in 47.5% of the population, a high prevalence compared with reports from developed countries. T. canis ova were positive in 43.6% of fecal samples from dog puppies. An endemic pattern of the disease is described: socioeconomic status, weathers, pollution, poor hygiene and a significant population of infected dogs. Neither the physical examination nor Elisa titers could detect any case of T. canis disease.

The ue of polysiloxane/polyvinyl alcohol beads as solid phase in IgG anti-Toxocara canis detection using a recombinant antigen

Immunodetection of human IgG anti-Toxocara canis was developed based on ELISA and on the use of polysiloxane/polyvinyl alcohol (POS/PVA) beads. A recombinant antigen was covalently immobilized, via glutaraldehyde, onto this hybrid inorganic-organic composite, which was prepared by the sol-gel technique. Using only 31.2 ng antigen per bead, a peroxidase conjugate dilution of 1:10,000 and a serum dilution of 1:200 were adequate for the establishment of the procedure. This procedure is comparable to that which utilizes the adsorption of the antigen to conventional PVC plates. However, the difference between positive and negative sera mean absorbances was larger for this new glass based assay. In addition to the performance of the POS/PVA bead as a matrix for immunodetection, its easy synthesis and low cost are additional advantages for commercial application.

In vitro and in vivo effects of Enterococcus faecalis CECT7121 on Toxocara canis

The aim of the present paper was to evaluate the larvicidal effect of Enterococcus faecalis CECT7121 (Ef7121) on the Toxocara canis cycle both in vitro and in vivo. For the in vitro experiments, T. canis larvae were incubated with the supernatants of Ef7121 (EI) and mutant Ef7121 (EIm), in a pre-culture of Ef7121 (EII) and in a fresh culture with Ef7121 (EIII) and the Ef7121 mutant strain (EIIIm). The viability of the larvae was calculated after a 48 h incubation. A significant reduction of the viability of T. canis larvae was observed in EI, EII and EIII. A decrease of this inhibitory effect was observed in EIm and EIIIm (p = 0.008). In the in vivo experiments, mice were orally inoculated with three doses of Ef7121. To study the probiotic persistence in the intestine, the animals were sacrificed every four days and their intestines were dissected. The initial average bacterial levels were 9.7 x 10(4) for Ef7121 (colony forming units/g). At the end of the assay the levels were 1.46 x 10(4). No bacterial translocation was detected in mesenteric lymphatic nodules and spleen. Ef7121 interference with the biological cycle was evaluated in mice challenged with T. canis. The interference was significant when the mice were challenged with probiotic and T. canis simultaneously (p = 0.001), but it was not significant when the challenge was performed 15 days after administration of the bacterial inoculum (p = 0.06). In conclusion, Ef7121 possessed in vitro and in vivo larvicidal activity.

Identification of candidate antigens from adult stages of Toxocara canis for the serodiagnosis of human toxocariasis

In the present work, we identified adult Toxocara canis antigens through sodium dodecyl sulfate-polyacrylamide gel electrophoresis for potential use in human toxocariasis immunodiagnosis. The sensitivity and specificity of several semi-purified antigens, as well as their cross-reactivity with other parasitic infections, were assessed by IgM and IgG-enzime linked immunosorbent assay. Whilst we found that the crude extract of the parasite presented limited sensitivity, specificity and high cross-reactivity against other parasites, we identified 42, 58, 68 and 97-kDa semi-purified antigens as the most promising candidates for immunodiagnosis. Moreover, the 58 and 68-kDa antigens presented the lowest IgM cross-reactivity. When tested as a combination, a mixture of the 58 and 68-kDa antigens presented 100% sensitivity and specificity, as well as minor cross-reactivity. Although the combination of the 42, 58, 68 and 97-kDa antigens presented 100% sensitivity at a dilution of 1:40, the low specificity and high cross-reactivity observed suggested a limited use for diagnostic purposes. Our data suggested that the 58 and 68-kDa antigens might be most suitable for the immunodiagnosis of human toxocariasis.

An evaluation of the dot-ELISA procedure as a diagnostic test in an area with a high prevalence of human Toxocara canis infection

The aim of this work was to evaluate a dot-enzyme-linked immunosorbent assay (dot-ELISA) using excretory-secretory antigens from the larval stages of Toxocara canis for the diagnosis of toxocariasis. A secondary aim was to establish the optimal conditions for its use in an area with a high prevalence of human T. canis infection. The dot-ELISA test was standardised using different concentrations of the antigen fixed on nitrocellulose paper strips and increasing dilutions of the serum and conjugate. Both the dot-ELISA and standard ELISA methods were tested in parallel with the same batch of sera from controls and from individuals living in the problem area. The best results were obtained with 1.33 µg/mL of antigen, dilutions of 1/80 for the samples and controls and a dilution of 1/5,000 for the anti-human IgG-peroxidase conjugate. All steps of the procedure were performed at room temperature. The coincidence between ELISA and dot-ELISA was 85% and the kappa index was 0.72. The dot-ELISA test described here is rapid, easy to perform and does not require expensive equipment. Thus, this test is suitable for the serological diagnosis of human T. canis infection in field surveys and in the primary health care centres of endemic regions.

Toxocara canis and the allergic process

The protective effect of infectious agents against allergic reactions has been thoroughly investigated. Current studies have demonstrated the ability of some helminths to modulate the immune response of infected hosts. The objective of the present study was to investigate the relationship between Toxocara canis infection and the development of an allergic response in mice immunised with ovalbumin (OVA). We determined the total and differential blood and bronchoalveolar lavage fluid cells using BALB/c mice as a model. To this end, the levels of interleukin (IL)-4, IL-5 and IL-10 and anti-OVA-IgE were measured using an ELISA. The inflammatory process in the lungs was observed using histology slides stained with haematoxylin and eosin. The results showed an increase in the total number of leukocytes and eosinophils in the blood of infected and immunised animals at 18 days after infection. We observed a slight lymphocytic inflammatory infiltrate in the portal space in all infected mice. Anti-OVA-IgE levels were detected in smaller proportions in the plasma of immunised and infected mice compared with mice that were only infected. Therefore, we concluded that T. canis potentiates inflammation in the lungs in response to OVA, although anti-OVA-IgE levels suggest a potential reduction of the inflammatory process through this mechanism.