Changes in hemocytes of Biomphalaria glabrata infected with Echinostoma paraensei and exposed to glyphosate-based herbicide.

The immune system of snails is highly sensitive to pollutants, which can suppress its immune response. We investigated the effects of exposure to the glyphosate-based herbicide Roundup® Original on the snail Biomphalaria glabrata infected by the platyhelminth Echinostoma paraensei by evaluating changes in the snail's internal defense system. Four cohorts were studied: control group, infected snails, snails treated with Roundup®, and snails infected and treated with Roundup®. The hemocyte viability was assessed, morphological differentiation of cells was observed and flow cytometry was performed to determine the morphology, viability and the lectin expression profiles. The frequencies of dead hemocytes were lower in the infected group and higher in both pesticide treated groups. Three cell types were identified: blast-like cells, hyalinocytes and granulocytes. The highest number of all types of hemocytes, as well as the highest number of dead cells, were observed in the infected, pesticide-treated group. The association between infection and herbicide exposure greatly increased the frequency of dead hemocytes, suggesting that this condition impairs the internal defense system of B. glabrata making the snails more vulnerable to parasitic infections.

In vivo and in vitro effects of the herbicide Roundup(®) on developmental stages of the trematode Echinostoma paraensei.

The exposure of wildlife and humans to toxic residues of Roundup(®) through agricultural practices or the food chain has been reported since the herbicide was found contaminating rivers. Glyphosate, N-(phosphonomethyl)glycine acid, is a nonselective post-emergent herbicide and is formulated as an isopropylamine salt with the surfactant taloamine polyethoxylate (POEA) representing the commercial formulation of Roundup(®). There is little knowledge about the effects of the herbicide on helminth parasites, particularly those whose life cycle is related to water bodies. Here we investigated the effects of the Roundup(®) on the food-borne trematode Echinostoma paraensei in experimental conditions using different developmental stages (eggs, miracidia, cercariae, metacercariae, newly excysted larvae (NEL), helminths at seven days and helminths at fourteen days). Three different herbicide concentrations were tested based on concentrations typically applied in the field: 225, 450 and 900 mg/L. Specimens were analyzed in vitro for hatching miracidia, mortality and excystment rate of metacercariae and in vivo for parasitic load and egg production. There was a significant difference in the hatching miracidia rate only for the newly embryonated eggs. The mortality of specimens and excystment rate of metacercariae were concentration-dependent. There was a significant difference in the miracidia mortality with respect to concentration until 56.3 mg/L. The same effect was observed for cercariae, and mortality was observed from 15 min onwards at concentrations of 225-900 mg/L. At low concentrations, mortality was detected after 30 min. The effects of the herbicide concentration on NEL and on helminths at seven and fourteen days showed a significant difference after 24 h. There was no significant difference in parasitic load and egg production after infection of rodents with exposed metacercariae. All developmental stages of the trematode E. paraensei were affected by Roundup(®) exposure under experimental conditions. These results suggest that dynamics of transmission of the trematode could be affected in the natural environments. The study also reinforces the usefulness of this trematode as a good model organism to test pesticides regarding human and environmental health.

The influence of exposure to Euphorbia splendens var. hislopii latex on the concentrations of total proteins and nitrogen products in Biomphalaria glabrata infected with Schistosoma mansoni.

The aim of this work was to analyze the content of total protein and nitrogen degradation products in Biomphalaria glabrata infected with Schistosoma mansoni and exposed to Euphorbia splendens var. hislopii latex. The LC(50) of this latex was 1.0mg/l. Concentrations of uric acid, urea and total proteins were determined in the hemolymph of B. glabrata infected with five S. mansoni miracidia and exposed to a sublethal concentration of E. splendens var. hislopii latex for 24h. The exposure to this molluscicide caused total depletion of the alterative sources of energy (total proteins) and significant variation in the nitrogen degradation products. The urea content increased while the uric acid level decreased. These results reflect a disturbance in the snails regulation of their metabolism due to intoxication caused by the latex exposure.

[Intestinal parasitic helminths in institutionalized dogs of Rio de Janeiro, Brazil]. / Parasitas gastrointestinais em cães institucionalizados no Rio de Janeiro, RJ.

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.

Molluscicidal action of the latex of Euphorbia splendens var. hislopii N.E.B. ("Christ's Crown") (Euphorbiaceae) against Lymnaea columella (Say, 1817) (Pulmonata: Lymnaeidae), intermediate host of Fasciola hepatica Linnaeus, 1758 (Trematode: Fasciolidae): 1- test in laboratory.

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.

Activity of Euphorbia splendens var. hislopii N.E.B. (Euphorbiaceae) latex against Lymnaea columella (Say, 1817) (Pulmonata: Lymnaeidae), intermediate host of Fasciola hepatica, Linnaeus, 1758 (Trematoda: Fasciolidae). 2: limited field-testing.

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.