RESUMO

The natural phytochemical latex of Euphorbia milii var. hislopii is one of the most promising natural molluscicides for the control of Biomphalaria glabrata, and has been widely studied under laboratory conditions for selective control of schistosomiasis transmission. However, the effect of this product on B. glabrata infected by other helminths had not yet been investigated. The present study reports evaluation of the effect of E. milii var. hislopii latex on the survival and reproductive activity of B. glabrata infected by Angiostrongylus cantonensis. For this purpose, the following groups were formed: control (C), exposed (E), infected (I) and infected and exposed for different time intervals (1 day - I+E-1D, 7 days - I+E-7D, 14 days - I+E-14D, 21 days - I+E-21D and 28 days - I+E-28D). The experimental infection was performed with L1 larvae of A. cantonensis and exposure to 0.08 mg/L (LC50) of E. milii latex for a period of 24 hours. We analyzed the effects of E. milii latex on the survival of snails during four weeks, reproductive parameters and possible histophysiopathological changes in the gonad and albumen gland of the snails. Regarding survival, there was reduction of more than 50% in the groups exposed to latex (E and I + E) compared to the control group. As for the number of ovigerous masses, eggs, and average number of hatched snails, significant increases were observed in the I+E-1D group at the 4th week compared to the control group and the other weeks in the same group. Angiostrongylus cantonensis larvae were observed in the gonad and albumen gland from day 21 and 28 of infection in groups I and I+E, respectively, with granuloma-like formation. At these observation periods and in these groups, an increase in galactogen was observed in the albumen gland, which influenced egg laying, suggesting the existence of a fecundity compensation mechanism phenomenon. It was possible to conclude that both stressors - A. cantonensis infection and exposure to E. milii latex - directly influenced the survival and reproductive parameters of B. glabrata.

O fitoquímico natural látex de Euphorbia milii var. hislopii é um dos mais promissores moluscicidas naturais para o controle de Biomphalaria glabrata, tendo sido amplamente estudado em condições laboratoriais como controle seletivo da transmissão da esquistossomose. No entanto, o efeito deste produto em B. glabrata infectada por outro helminto, ainda não tinha sido investigado. O presente trabalho tem por objetivo avaliar o efeito do látex de E. milii var. hislopii na sobrevivência e atividade reprodutiva de B. glabrata infectada por Angiostrongylus cantonensis. Para isso, foram formados os seguintes grupos: controle (C), exposto (E), infectado (I) e infectado e exposto em diferentes intervalos de tempo de infecção (1 dia - I+E-1D, 7 dias - I+E-7D, 14 dias - I+E-14D, 21 dias - I+E-21D e 28 dias - I+E-28D). A infecção experimental foi realizada com larvas L1 de A. cantonensis e a exposição com 0,08 mg/L (LC50) do látex de E. milii por um período de 24 horas. Foram analisados os efeitos do látex de E. milii quanto à sobrevivência dos moluscos ao longo de quatro semanas, aos parâmetros reprodutivos e as possíveis alterações histofisiopatológicas na gônada e na glândula de albúmen dos moluscos analisados. Em relação à sobrevivência foi possível observar redução de mais de 50% nos grupos expostos ao látex (E e I + E) em comparação ao grupo controle. Quanto ao número de massas ovígeras, ovos e número médio de moluscos eclodidos pode-se observar um aumento significativo no grupo I+E-1D na 4ª semana em comparação ao grupo controle e as demais semanas no mesmo grupo. Larvas de A. cantonensis foram observadas na glândula e na glândula de albúmen, a partir dos 21º dias e 28º dia de infecção nos grupos I e I+E, respectivamente, com a formação de granuloma-like. Nesses períodos de observação e nesses grupos pode-se observar na glândula de albúmen, um aumento do conteúdo de galactogênio, o que influenciou a postura dos ovos, sugerindo um fenômeno de compensação da fecundidade. Foi possível concluir que ambos agentes estressores, infecção por A. cantonensis e exposição ao látex de E. milii influenciou diretamente a sobrevivência e os parâmetros reprodutivos de B. glabrata.

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Plant molluscicides have been regarded as possible alternatives to the costly and environmentally hazardous molluscicides currently available. This study was undertaken to compare the developmental toxicity of a plant molluscicide (Euphorbia milii latex, LAT) with that of three synthetic molluscicidal compounds. Biomphalaria glabrata egg masses (0-15 h after spawning) were exposed to molluscicides for 96 h and thereafter examined up to the 14th day after spawning. Embryo deaths, abnormal embryo development (malformations) and the day of hatching were recorded. Although exhibiting a weak ovicidal effect, LAT markedly impaired the development of snail embryos at concentrations 1000 microg L(-1) and produced anomalies (EC(50)=2040 microg L(-1)) such as abnormal shells, hydropic embryos, cephalic and non-specific malformations. Embryolethal potencies of molluscicides were as follows: triphenyltin hydroxide (TPTH; LC(50)=0.30 microg L(-1))>niclosamide (NCL; LC(50)=70 microg L(-1))>copper sulphate (CuSO(4); LC(50)=2190 microg L(-1)) >>> LAT (LC(50)=34030 microg L(-1)). A few malformations were recorded in embryos exposed to concentrations of TPTH within the range of lethal concentrations, while almost no anomalies were noted among those treated with NCL or CuSO(4). A hatching delay (hatching on day 10 after spawning or later) was observed among LAT-exposed embryos. The effects of NCL, TPTH and CuSO4 on hatching were to some extent masked by their marked embryolethality. The no-observed effect concentrations (NOEC) for embryotoxicity were as follows: TPTH, 0.1 microg L(-1); NCL, 25.0 microg L(-1); CuSO(4), 500.0 microg L(-1) and LAT, 500.0 microg L(-1). Results from this study suggest that, although LAT was not acutely embryolethal after a short-term exposure, it markedly disrupted snail development. The marked embryotoxicity of E. milii possibly contributes to its effectiveness as a molluscicide.

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Gastrópodes pulmonados terrestres podem atuar como hospedeiros intermediários de helmintos. Os primeiros registros do controle químico desses invertebrados datam do início do século XX e as substâncias utilizadas eram toxinas inespecíficas já empregadas no controle de outras pragas. Moluscicidas sintéticos apresentam limitações técnicas que estimularam a busca de substitutos naturais. Dentre as várias espécies vegetais com atividade moluscicida, Euphorbia cotinifolia L., Euphorbia milii des Moul. var. splendens (Bojer ex Hook) Ursch & Leandri e Euphorbia tirucalli L. despertam atenção pelos excelentes resultados obtidos sobre moluscos aquáticos. Contudo, estudos sobre a atividade de plantas moluscicidas em moluscos terrestres são pouco comuns, apesar de sua grande importância parasitológica e agrícola. As semelhanças anatomo-fisiológicas entre espécies de moluscos aquáticos e terrestres sugerem que estratégias de controle químico possam ter eficiência semelhante para os dois grupos de invertebrados. Com base nessa hipótese, o presente trabalho teve como objetivo avaliar a atividade moluscicida do látex de três espécies do gênero Euphorbia sobre Leptinaria unilamellata d'Orbigny, 1835, gastrópode terrestre descrito como hospedeiro intermediário de helmintos trematódeos digenéticos que parasitam animais domésticos. Destas E. milii var. splendens apresentou efeito moluscicida elevado sobre L. unilamellata, 100 por cento até uma diluição de 1:800, já nos primeiros minutos após a aplicação. Embora citadas na literatura como tóxicas para moluscos aquáticos, E. cotinifolia e E. tirucalli não exibiram atividade moluscicida sobre L. unilamellata. Os resultados do presente estudo indicam que o látex de E. milii var. splendens pode se constituir em uma estratégia viável de controle químico de moluscos terrestres.

Pulmonate terrestrial gastropods can be intermediate hosts for helminthes. The first records of chemically controlling these invertebrates date back to the beginning of the 20th century. The substances used were unspecific toxins which were already used for controlling other pests. Synthetic molluscicides have technical limitations that stimulated a search for natural substitutes. Among the many vegetal species that have molluscicidal activity, Euphorbia cotinifolia L., Euphorbia milii des Moul. var. splendens (Bojer ex Hook) Ursch & Leandri and Euphorbia tirucalli L. call attention because of the excellent results that have been obtained when they are used on aquatic mollusks. However, studies on the activity of molluscicidal plants on terrestrial mollusks are rare, in spite of its great importance in controlling parasites and to agriculture. Anatomical and physiological similarities among species of aquatic and terrestrial mollusks suggest that chemical control strategies can be effective on both groups of invertebrates. Based on this hypothesis, we assessed the molluscicidal activity of the latex of three species of the genus Euphorbia on the terrestrial gastropod Leptinaria unilamellata d'Orbigny, 1835, described as an intermediate host of digenetic trematode helminths that parasitize domestic animals. E. milii var. splendens had the highest molluscicidal effect on L. unilamellata, being 100 percent lethal in concentrations up to 1:1800 in the first minutes after application. Although E. cotinifolia and E. tirucalli have been cited in the literature as toxic to aquatic mollusks, they did not have molluscicidal activity on L. unilamellata. The results of our study indicate that the latex of E. milii var. splendens may be a viable strategy for the chemical control of terrestrial mollusks.

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Four adult sexually matured and clinically healthy West African Dwarf (WAD) rams aged between 24 and 30 months were used for the study. The rams were first used as control and later as experimental animals upon being orally dosed with Euphorbia hirta extract at 400mg/kg body weight for 14 days. Semen samples were collected from the rams a day after the administration of the plant extra and seven days after. The objective of the study was to investigate the effect of Euphorbia hirta on the semen picture of WAD rams. There were significantly differences (P <0.05) in the semen picture as reflected in a reduction of sperm motility from 80% to 47.5% and live-dead ratio from 90.75% to 32.5% in the control and post-experimental stages of the study respectively. This indicates that the fertilization capacity and livability of spermatozoa were negatively affected. There were no significant differences in the values of body parameters measured across the stages of the study. The plant is therefore not recommended for medicinal purpose in male animals.

Cuatro carneros enanos adultos de África Occidental sexualmente maduros y clínicamente sanos, con edades comprendidas entre los 24 y 30 meses, fueron utilizados para este estudio. Los carneros fueron utilizados como control y, más tarde, como animales de experimentación al ser medicados por vía oral con extracto de Euphorbia hirta en 400mg/kg peso corporal durante 14 días. Se recogieron muestras de semen de los carneros un día después de la administración de la planta y siete días después. El objetivo del estudio fue investigar el efecto de Euphorbia hirta en las imágenes de esperma de carneros enanos África Occidental. Hubo diferencias significativas (P <0,05) en la imagen del semen como reflejo de una reducción de la motilidad espermática del 80% al 47,5% y un ratio de vivos-muertos de 90,75% a 32,5% en la etapa control y después de las fases experimentales del estudio, respectivamente. Esto indica que la capacidad de fertilización y calidad de vida de los espermatozoides fueron afectados negativamente. No hubo diferencias significativas en los valores de los parámetros corporales medidos a través de las etapas del estudio. La planta por tanto no es recomendable para fines medicinales en los animales machos.

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