Resumo Helmintos parasitos tem sido estudados como acumuladores potenciais para diferentes poluentes. O trematódeo E. paraensei tem como hospedeiro vertebrado o roedor Nectomys squamipes naturalmente exposto a pesticidas no meio ambiente. No entanto, pouca informação está disponível sobre os efeitos dos pesticidas em helmintos parasitos. O presente estudo investigou, em condições experimentais, os efeitos morfológicos no trematódeo E. paraensei após a exposição ao herbicida Roundup®, em concentrações abaixo das recomendadas para a utilização agrícola. A microscopia eletrônica de varredura (MEV) mostrou após duas horas de exposição, alterações no tegumento, como enrugamento, contração e descamação com perda de espinhos no colar peristômico e análise histopatológica evidenciou células do ceco alteradas, as glândulas vitelínicas com vacúolos e mudanças estruturais nas camadas musculares. Diminuição do conteúdo glicídico, principalmente no tecido conjuntivo, também foi observado. Considerando a predisposição à exposição a pesticidas agrícolas de N. squamipes infectado por E. paraensei, são necessários estudos para avaliar o impacto de tais resíduos frente aos helmintos e seus hospedeiros.

Abstract Helminth parasites have been studied as potential accumulators for different pollutants. Echinostoma paraensei is a foodborne trematode whose vertebrate host, the rodent Nectomys squamipes, is naturally exposed to environmental pesticides. However, little information exists regarding the pesticide’s effects on helminths. This study investigated the morphological effects on the trematode, E. paraensei, after experimental Roundup® herbicide exposure, in concentrations below those recommended for agricultural use. After two hours of exposure, scanning electron microscopy (SEM) showed changes to the tegument, such as furrowing, shrinkage, peeling, spines loss on the peristomic collar, and histopathological evidence of altered cells in the cecum and acinus vitelline glands with vacuoles and structural changes to the muscular layers. Glycidic content was decreased, primarily in the connective tissue. As E. paraensei is an intestinal parasite of the semi-aquatic wild rodent, N. squamipes, it is predisposed to pesticide exposure resulting from agricultural practices. Therefore, we emphasize the need to evaluate its impact on helminth parasites, due to their pivotal role in regulating host populations.

This paper evaluates the alterations in the glycogen content of tissues (digestive gland and cephalopedal mass) and glucose in the haemolymph of Biomphalaria glabrata BH strain infected with Schistosoma mansoni BH strain and exposed to the latex of Euphorbia splendens var. hislopii. A reduction in the glycogen deposits was observed in infected snails exposed and not exposed to latex. However, the exposure to latex caused a greater depletion of the glycogen levels in both sites analysed, especially from the third week onward. The utilisation of latex as a molluscicide to control the population of infected B. glabrata selectively is proposed.

O método complexométrico é geralmente utilizado para determinação quantitativa de cálcio em diferentes materiais; contudo a aplicação deste método à determinação de cálcio em conchas de moluscos infere significativas interferências aos resultados obtidos. O molusco Bradybaena similaris, um gastrópode terrestre, foi utilizado como modelo para aperfeiçoar este método. As conchas foram calcinadas e dissolvidas em ácido nítrico, o peróxido de hidrogênio também foi empregado para clarificar o meio após a adição do ácido. O processo de calcinação e o uso do ácido nítrico reduziram significativamente as interferências, permitindo um maior grau de destruição das substâncias orgânicas da concha. O aperfeiçoamento da técnica para determinação de cálcio empregado resultou em uma quantidade maior de cálcio igual a 874.24 ± 56.617 mg de CaCO3/g de cinzas em comparação com a técnica convencional que permitiu a determinação de 607.79 ± 67.751 mg de CaCO3/g de concha, peso fresco.

The complexometric method is usually applied to quantitative calcium determination in different materials; however the application of this method to calcium determination in molluscs shells infers significant interferences to the results. The snail Bradybaena similaris, a terrestrial gastropod, was used as experimental model to the improvement of this method. The shells were calcinated and dissolved in nitric acid, the hydrogen peroxide was also used to clarify the medium after the acid addition. The calcination procedure and the use of nitric acid reduced the significantly the interferences, allowing a major degree of destruction of the organic substances of the shell. The improvement of the calcium determination technique usually employed showed calcium content of 874.24 ± 56.617 mg of CaCO3/g of ash in comparison to the conventional technique that allowed the determination of 607.79 ± 67.751 mg of CaCO3/g of shell, wet weight.

The reproductive activity of Biomphalaria glabrata exposed to Euphorbia splendens var. hislopii latex was evaluated. Parameters related to fecundity and fertility were observed. The snails were exposed to the LD50 (1 mg/l) of crude latex. At the first week post exposure (p.e.), the egg laying was reduced. After the fourth week p.e., an increase of the number of eggs/snail occurred. The results showed a marked reduction in the hatching of the snails, revealing an interference of latex exposure with the reproductive process of B. glabrata of E. splendens var. hislopii. The LD50 of the latex may be used as an alternative method to control the size of the populations of B. glabrata in field.

Foi estudada a ocorrência de parasitas gastrointestinais em cães recolhidos e mantidos em instituto público de medicina veterinária no Rio de Janeiro, RJ. Amostras de fezes frescas foram coletadas em março de 2004 e analisadas pelos métodos de flutuação de Willis e centrífugo-flutuação em solução de sacarose. De 204 amostras, 45,6% estavam positivas para helmintos gastrointestinais.

The occurrence of intestinal parasitic helminths in dogs from a public institute of veterinary medicine in Rio de Janeiro was assessed. Samples of fresh feces were collected in March 2004 and analyzed by Willis and centrifugal-flotation in sucrose solution methods. Of 204 samples, 45.6% were positive for helminths.

Foi estudada a ocorrência de parasitas gastrointestinais em cães recolhidos e mantidos em instituto público de medicina veterinária no Rio de Janeiro, RJ. Amostras de fezes frescas foram coletadas em março de 2004 e analisadas pelos métodos de flutuação de Willis e centrífugo-flutuação em solução de sacarose. De 204 amostras, 45,6% estavam positivas para helmintos gastrointestinais.

The occurrence of intestinal parasitic helminths in dogs from a public institute of veterinary medicine in Rio de Janeiro was assessed. Samples of fresh feces were collected in March 2004 and analyzed by Willis and centrifugal-flotation in sucrose solution methods. Of 204 samples, 45.6% were positive for helminths.

Molluscides have been used as one of the strategies to control schistosomiasis. Many plant extracts with molluscidal effects have been tested, but the action of the latex of Euphorbia splendens var. hislopii is considered the most promising because it meets the recommendations of the World Health Organization (WHO). The objective of this study was to determine the lethal dose and identify the effects of the different doses of latex of E. splendens var. hislopii on the physiology of Biomphalaria glabrata submitted to treatment for 24 h. The concentrations of glucose, uric acid and total proteins in the hemolymph and of glycogen in the digestive gland and cephalopodal mass were determined. The LD50 value was 1 mg/l. The highest escape index was found to be at a concentration of 0.6 mg/l. The results showed that the latex of E. splendens var. hislopii caused a sharp reduction in the reserves of glycogen in the digestive gland and elevation of the protein content in the hemolymph of B. glabrata.

We report the first finding of Lymnaea cousini naturally infected with Fasciola hepatica in Ecuador. A sample of 70 snails was collected in April 2005 from a wetland located in a valley at approximately 3000 m a.s.l., near the locality of Machachi, Pichincha Province. The prevalence of natural infection in L. cousini was 31.43%, which is the highest value ever recorded for naturally infected lymnaeid species.

The molluscicidal evaluation of Euphorbia splendens var. hislopii (Crown of thorns) against Lymnaea columella snails, intermediate host of Fasciola hepatica, in irrigation ditches of the Pisciculture Station at Universidade Federal Rural do Rio de Janeiro, was studied under limited field conditions. An aqueous solution of the latex at 5 mg/l was tested in two irrigation ditches (experimental and control ditches), after initial sampling of the snail population present. Twenty-four hours after application of the product, it was verified that 97.4% of free L. columella snails and 100% of snails of the same species captive in cages and used as sentinels at three points equidistant from the application site in the experimental ditch, died. For Biomphalaria tenagophila and Melanoides tuberculata snails, present in the experimental ditch, the mortality was 100%, for the species Pomacea spp. the mortality was 40%. No mortality was verified in the free mollusks, or in the sentinels in the ditch used as control. E. splendens var. hislopii latex is thus an efficient natural molluscicide, which may be used as an alternative control agent against L. columella.

Bioensaios de laboratório e de campo têm comprovado a especificidade da ação moluscicida do látex da Euphorbia splendens var. hislopii (coroa-de-Cristo) (Euphorbiaceae) para os moluscos das espécies Biomphalaria glabrata, B. tenagophila, B. straminea, B. pfeifferi e Bulinus sp. No presente estudo foi investigado o efeito da variação do pH na concentração letal (CL90) testada para B. tenagophila. Bioensaios com soluções aquosas do látex variando de 0,4 a 12,0 µl/l foram ajustadas aos valores de pH 5,0; 6,0; 7,0 e 8,0, e testadas de acordo com os métodos padronizados pela Organização Mundial da Saúde. Os resultados obtidos indicam que a menor concentração letal do látex ocorreu em pH 6,0 (CL90 = 3,2 µl/l) e a maior em pH 8,0 (LC90 = 10,3 µl/l). As concentrações letais (LC90) ajustadas em pH 5,0 e 7,0 foram 3,4 µl/l e 4,7 µl/l, respectivamente. A partir destes dados conclui-se que a toxicidade do moluscicida não se alterou quando as concentrações foram ajustadas em pH 5,0 e 6,0, pois observou-se que a mortalidade foi de 100% a partir da concentração de 2,0 µl/l, o que não ocorreu nas concentrações com ajuste de pH 7,0 e 8,0.

Laboratory and field bioassays have confirmed the specificity of the molluscicidal activity of the Euphorbia splendens var. hislopii latex (crown of Christ) (Euphorbiaceae) over snails of the species Biomphalaria glabrata, B. tenagophila, B. straminea, B. pfeifferi and Bulinus sp. in the control of Schistosoma mansoni. In the present study, the effect of the pH variation on lethal concentration (LC90) over B. tenagophila was evaluated. Bioassays with the aqueous solutions of the latex ranging from 0.4 to 12 µl/l were adjusted for pH of 5.0; 6.0; 7.0 and 8.0, and tested in accordance with methods standardized by World Health Organization. The results obtained indicated that the minor concentration of the latex occurred at pH 6.0 (LC90 = 3.2 µl/l) and the maximum at pH 8.0 (LC90 = 10.3 µl/l). Lethal concentrations adjusted for pH 5.0 and 7.0 were 3.4 µl/l and 4,7µl/l, respectively. From the results it could be concluded that the molluscicidal toxicity was not altered when the concentrations were adjusted for pH 5.0 and 6.0, as we observed that mortality rate was 100% starting at a concentration of 2.0 µl/l, not the same for the concentrations with adjustment for pH 7.0 and 8.0.

The latex action of Euphorbia splendens var. hislopii (Christ's Crown) against snails Lymnaea columella, intermediate host of Fasciola hepatica, derived from irrigation ditches of the Station of Pisciculture at Universidade Federal Rural do Rio de Janeiro, was studied in the laboratory. Lab bioassays, using aqueous solutions of the latex, varying between 0.1 and 10 mg/l, have proven molluscicidal activity of the product collected on the same day the tests were performed, during the four seasons of the year, finding the following lethal concentrations (LC90): 1.51 mg/l in the spring; 0.55 mg/l in the summer; 0.74 mg/l in the fall and 0.93 mg/l in winter, after 24 h exposure of the snails, showing significant differences among the seasons of the year (ANOVA test, F = 11.01, G.L.= 3/33, p < 0.05), as well as among the concentrations (ANOVA test, F = 27.38, G.L.= 11/33, p < 0.05). In the summer, mortality reached 100% from concentration at 0.6 mg/l, the same during fall and in winter as of 1 mg/l, while in spring it only reached 100% mortality as of 2 mg/l. Mortality in the controls was low, reaching 5% in the summer and winter and 10% in the fall and spring. None of the samples died. During the assay, with an aqueous solution of the latex at a concentration of 5 mg/l, in order to check the time of duration of the product effect, in the laboratory, it was observed that the molluscicidal activity remained stable up to the 15th day after the beginning of the test with 100% mortality of L. columella, gradually losing its effect until the 23rd day, when we no longer observed animal mortality. In the control group, there was a random daily variation in mortality rate ranging 0-50% after 48 h of observation for 30 days.

The aim of this study was to determine the toxicity of niclosamide (Bayluscide ®) on Melanoides tuberculata and Biomphalaria glabrata under laboratory conditions. The latter species is the intermediate host of Schistosoma mansoni (Sambon 1917). M. tuberculata was successfully used as competitor of B. glabrata in biological control programs in French West Indies. Both molluscicide and biological control using M. tuberculata have proved to be successful in reducing the population density of B. glabrata. The associated use of molluscicide in this area would be an effective measure if M. tuberculata were less susceptibility to the molluscicide than B. glabrata. Three hundreds individuals each of B. glabrata and of M. tuberculata, collected in Sumidouro, State of Rio de Janeiro, were used in the experiment. The molluscs were exposed to 14 different concentrations of niclosamide as recommended by the World Health Organization. Probit analysis was used to determine the LC 50 and LC 90. The LC 50 and LC 90 values for B. glabrata were 0.077 mg/l and 0.175 mg/l, respectively and the LC 50 and LC 90 values for M. tuberculata were 0.082 mg/l and 0.221 mg/l respectively. As the lethal concentrations of niclosamide were approximately the same to both species, this could be a disadvantage when controlling B. glabrata with niclosamide in an area of M. tuberculata occurrence. It migth therefore be preferable to utilize the latex extracted from the Euphorbia splendens, which presented a much higher efficiency for B. glabrata than to M. tuberculata.

Foi realizada a infecção experimental de Physa cubensis Pfeiffer, 1839, com miracídios de Fasciola hepatica. Para tanto, cada um dos cem moluscos, selecionados para o experimento, foram infectados com três miracídeos. Alguns exemplares, escolhidos ao acaso, foram mantidos até o 60º dia para observação da emergência de formas larvais. Os moluscos restantes foram fixados após 0,5; 1; 2; 3; 4; 5; 24; 48; 72; 96; 120 e 144 horas de infecção, utilizando Raillet & Henry. O trabalho foi realizado de forma comparativa com Lymnaea columella. Os resultados demonstraram o desenvolvimento das formas larvais do trematódeo em L. columella e a presença de esporocistos em vários estágios de degeneração na região cefalopodal e na região do manto nos primeiros dias de infecção em P. cubensis.

The experimental infection of Physa cubensis Pfeiffer, 1839 was realized with three Fasciola hepatica miracidiae. One hundred snails were infected. Some groups were maintained up to the 60º. day post infection for the observation of the larval forms emergency. The snails, in the remaining groups, were immerged in Raillet & Henry 0,5; 1; 2; 3; 4; 5; 24; 48; 72; 96; 120 and 144 hours post infection. The experiment was realized comparatively with Lymnaea columella. The results demonstrated, at first days infection, esporocysts presence in the cephalopodal mass and mantle, however, in the last days it didn't get to identify any larval form, in P. cubensis.

Observações histológicas em moluscos da espécie Lymnaea columella foram realizadas para determinar a existência ou não de alterações provocadas pelo uso do látex de Euphorbia splendens var. hislopii. O experimento foi realizado de acordo com os métodos padronizados pela OMS. Cinqüenta exemplares de L. columella, com comprimento de concha de 6±1mm, foram submetidos a 200ml das soluções nas concentrações de 0,47µl/L; 0,76µl/L; 0,94µl/L e 1,51µl/L, durante 24 horas. Após o evento experimental, os moluscos foram colocados em solução de Raillet & Henry, para sacrifício e fixação. Os resultados demonstraram lesões caracterizadas por degeneração, necrose e acúmulo de líquido na glândula digestiva e no rim, em exemplares submetidos a concentrações a partir de 0,47 mil/L.

Histological observations in Lymnaea columella snails to determine the existence or not of changes caused by the use of the Euphorbia splendens's latex were realized. The experiment was realized in agreement with the methods standardized by WHO. Fifty samples of L. columella, with shell 6±1mm long, were submitted at 200ml of the solutions in concentrations of 0,47µl/L; 0,76µl/L; 0,94µl/L and 1,51µl/L, during 24 hours. After the experimental event, the snails were placed in solution of Raillet & Henry, for sacrifice and fixation. The results demonstrated lesions characterized by degeneration, necrosis and liquid accumulation in the digestive gland and in the kidney, in samples maintained in concentrations from 0,47 mul/L on.

Registrou-se no município de Resende, Estado do Rio de Janeiro, Brasil, a primeira ocorrência do molusco Achatina fulica hospedeiro intermediário de Angiostrongylus cantonensis, causador da angiostrongilíase meningoencefálica. Em cinco bairros visitados, foram encontrados moluscos vivendo livremente, e nenhum dos animais coletados apresentava a forma larvar do parasito. A presença de A. fulica pode estar relacionada à comercialização desse molusco como alimento, e representa possibilidade de instalação dessa zoonose na região.

The first occurrence of the Achatina fulica, an intermediate host of Angiostrongylus cantonensis, the etiological agent of meningoencephalic angiostrongiliasis, is reported in Resende municipality, Brazil. In the five visited localities, snails were found living freely, and the larvae of this parasite was not seen in any of them. The finding of A. fulica in the area may be related to its commercialization as a food item and embodies the possibility of new focus.