Chemical composition and effect of essential oils of Thymus vulgaris and Origanum vulgare on adults and ovigerous masses of Pseudosuccinea columella.

Fasciolosis is a worldwide distribution zoonosis that causes great damage in ruminant breeding and has the aquatic mollusc Pseudosuccinea columella as an intermediate host. Synthetic molluscicides are the most used for control; however, they are harmful to fauna and flora. Therefore, this study aimed to evaluate the effect of essential oils from Thymus vulgaris, Origanum vulgare, and terpene carvacrol, on adult molluscs and eggs of P. columella. Analysis of EO volatile components was carried out on a gas chromatograph equipment coupled with mass spectrometry selective detector. The studied components were diluted in concentrations of 10, 20, 40, 60, 80 and 100 ppm, and it was observed that O. vulgare at concentrations of 60, 80 and 100 ppm, carvacrol at the concentrations of 80 and 100 ppm, and T. vulgaris at a concentration of 80 ppm led to 100% mortality of molluscs. All concentrations the substances tested showed 100% ovicidal activity.

Spread of the fascioliasis endemic area assessed by seasonal follow-up of rDNA ITS-2 sequenced lymnaeid populations in Cajamarca, Peru.

Fascioliasis is a worldwide emerging snail-borne zoonotic trematodiasis with a great spreading capacity linked to animal and human movements, climate change, and anthropogenic modifications of freshwater environments. South America is the continent with more human endemic areas caused by Fasciola hepatica, mainly in high altitude areas of Andean regions. The Peruvian Cajamarca area presents the highest human prevalences reported, only lower than those in the Bolivian Altiplano. Sequencing of the complete rDNA ITS-2 allowed for the specific and haplotype classification of lymnaeid snails collected in seasonal field surveys along a transect including 2007-3473 m altitudes. The species Galba truncatula (one haplotype preferentially in higher altitudes) and Pseudosuccinea columella (one haplotype in an isolated population), and the non-transmitting species Lymnaea schirazensis (two haplotypes mainly in lower altitudes) were found. Climatic seasonality proved to influence G. truncatula populations in temporarily dried habitats, whereas L. schirazensis appeared to be more climatologically independent due to its extreme amphibious ecology. Along the southeastern transect from Cajamarca city, G. truncatula and L. schirazensis shared the same site in 7 localities (46.7% of the water collections studied). The detection of G. truncatula in 11 new foci (73.3%), predominantly in northern localities closer to the city, demonstrate that the Cajamarca transmission risk area is markedly wider than previously considered. Lymnaea schirazensis progressively increases its presence when moving away from the city. Results highlight the usefulness of lymnaeid surveys to assess borders of the endemic area and inner distribution of transmission foci. Similar lymnaeid surveys are still in need to be performed in the wide northern and western zones of the Cajamarca city. The coexistence of more than one lymnaeid transmitting species, together with a morphologically indistinguishable non-transmitting species and livestock movements inside the area, conform a complex scenario which poses difficulties for the needed One Health control intervention.

Lymnaeidae from Santander and bordering departments of Colombia: Morphological characterization, molecular identification and natural infection with Fasciola hepatica.

The Lymnaeidae constitute a family of freshwater gastropod molluscs whose diversity and ecology have been infrequently studied throughout Colombia. Some lymnaeid species act as intermediate hosts of trematode parasites, which are of great importance in both the veterinary and medical fields. Among trematode parasites, Fasciola hepatica is best known for being an important parasite of sheep and cattle for decades and causes significant economic losses in these livestock species. The main objective of this work is to identify the various species of lymnaeids that occupy different geographical regions of Santander and its bordering departments within Colombia. This will expand the knowledge of lymnaeid diversity in Colombia and provide further insight into their role in the transmission of F. hepatica. A total of 118 georeferenced sites between 126 m.a.s.l. and 3870 m.a.s.l. were sampled in Santander, Boyacá, Norte de Santander and Cundinamarca, respectively. Lymnaeid snails were identified according to the morphology of their shells and by several characteristics of their reproductive systems. Species identification was confirmed using DNA barcoding. Four lymnaeid species are reported in the study area: the native Galba cousini and three exotic species, Pseudosuccinea columella, G. truncatula and G. schirazensis. The four species were examined for natural infection with F. hepatica. Infected variants of the main snail host, G. cousini, were found in the Onzaga, Encino and Vetas municipalities of Santander, as well as in the Belén municipality of Boyacá. A second species, G. truncatula was also found naturally infected in Mutiscua municipality of Norte de Santander. The two other species, P. columella and G. schirazensis were found free of infection.

Lymnaeid Snail Vectors of Fascioliasis, Including the First Finding of Lymnaea neotropica in Ecuador, Assessed by Ribosomal DNA Sequencing in the Southern Zone Close to the Peru Border.

PURPOSE: Fascioliasis is a freshwater snail-borne zoonotic trematodiasis of high pathogenicity and wide veterinary repercussions. In South America, moreover, it causes serious public health problems, with high human infection rates in Andean countries. Ecuador offers a worrying risky scenario due to its physiography, including many human infection reports and animal endemicity throughout its Andean highlands. METHODS: Endemic areas with increasing animal fascioliasis were surveyed for lymnaeid snails in the province of Loja, southern Ecuador, close to the border of Peru, the country known to present the widest human fascioliasis endemic zone. The altitude of the sampling sites ranged between 150 and 1770 m a.s.l., and their location was close to human villages. Biotopes surveyed were characterized according to fascioliasis transmission needs. RESULTS: The species Lymnaea schirazensis and L. neotropica were identified by rDNA ITS-2 and ITS-1 sequencing. The non-transmitting L. schirazensis combined haplotype agreed with populations of this species previously reported from northern Ecuador. The finding of the efficient vector L. neotropica is reported for the first time in Ecuador and suggests a passive introduction from neighbouring Peru by uncontrolled livestock transport. CONCLUSIONS: Rice irrigation system implementation, lymnaeid finding on Taraxacum (dandelion) plants which are consumed fresh in salads by people, and Saccharum (sugarcane), whose bark is peeled off with the teeth, represent potential infection sources for humans. The closeness to the Cajamarca human hyperendemic area in northern Peru, where the same two lymnaeids have been also found and triclabendazole resistance reported, is an additional risk to be considered regarding the livestock transborder exchange.

GABA-like immunoreactivity in Biomphalaria: Colocalization with tyrosine hydroxylase-like immunoreactivity in the feeding motor systems of panpulmonate snails.

The simpler nervous systems of certain invertebrates provide opportunities to examine colocalized classical neurotransmitters in the context of identified neurons and well defined neural circuits. This study examined the distribution of γ-aminobutyric acid-like immunoreactivity (GABAli) in the nervous system of the panpulmonates Biomphalaria glabrata and Biomphalaria alexandrina, major intermediate hosts for intestinal schistosomiasis. GABAli neurons were localized in the cerebral, pedal, and buccal ganglia of each species. With the exception of a projection to the base of the tentacle, GABAli fibers were confined to the CNS. As GABAli was previously reported to be colocalized with markers for dopamine (DA) in five neurons in the feeding network of the euopisthobranch gastropod Aplysia californica (Díaz-Ríos, Oyola, & Miller, 2002), double-labeling protocols were used to compare the distribution of GABAli with tyrosine hydroxylase immunoreactivity (THli). As in Aplysia, GABAli-THli colocalization was limited to five neurons, all of which were located in the buccal ganglion. Five GABAli-THli cells were also observed in the buccal ganglia of two other intensively studied panpulmonate species, Lymnaea stagnalis and Helisoma trivolvis. These findings indicate that colocalization of the classical neurotransmitters GABA and DA in feeding central pattern generator (CPG) interneurons preceded the divergence of euopisthobranch and panpulmonate taxa. These observations also support the hypothesis that heterogastropod feeding CPG networks exhibit a common universal design.

Human fascioliasis endemic areas in Argentina: multigene characterisation of the lymnaeid vectors and climatic-environmental assessment of the transmission pattern.

BACKGROUND: In South America, fascioliasis stands out due to the human endemic areas in many countries. In Argentina, human endemic areas have recently been detected. Lymnaeid vectors were studied in two human endemic localities of Catamarca province: Locality A beside Taton and Rio Grande villages; Locality B close to Recreo town. METHODS: Lymnaeids were characterised by the complete sequences of rDNA ITS-2 and ITS-1 and fragments of the mtDNA 16S and cox1. Shell morphometry was studied with the aid of a computer image analysis system. Climate analyses were made by nearest neighbour interpolation from FAO data. Koeppen & Budyko climate classifications were used. De Martonne aridity index and Gorczynski continentality index were obtained. Lymnaeid distribution was assessed in environmental studies. RESULTS: DNA sequences demonstrated the presence of Lymnaea neotropica and L. viator in Locality A and of L. neotropica in Locality B. Two and four new haplotypes were found in L. neotropica and L. viator, respectively. For interspecific differentiation, ITS-1 and 16S showed the highest and lowest resolution, respectively. For intraspecific analyses, cox1 was the best marker and ITS-1 the worst. Shell intraspecific variability overlapped in both species, except maximum length which was greater in L. viator. The desertic-arid conditions surrounding Locality A, the semiaridity-aridity surrounding Locality B, and the very low yearly precipitation in both localities, are very different from the typical fascioliasis transmission foci. Lymnaeids are confined to lateral river side floodings and small man-made irrigation systems. Water availability only depends on the rivers flowing from neighbouring mountains. All disease transmission factors are concentrated in small areas where humans and animals go for water supply, vegetable cultures and livestock farming. CONCLUSIONS: The unusually high number of DNA haplotypes and the extreme climate unsuitable for F. hepatica and lymnaeid development, demonstrate that the transmission foci are isolated. Seasonal transmission may depend on the timely overlap of appropriate temperature and river water availability. Lymnaeids and F. hepatica have probably reached these localities by livestock introduction. DNA differences regarding other populations of L. neotropica and L. viator in Argentina suggest an introduction independent from the spreading movements which allowed these two lymnaeids to expand throughout the country.

Lymnaea cousini , huésped de Fasciola hepatica en el trópico alto andino de Colombia, y sus nuevos haplotipos, confirmados con el marcador mitocondrial del gen de la citocromo oxidasa I / Lymnaea cousini , intermediate host of Fasciola hepatica in the Colombian high tropical Andes, and its new haplotypes confirmed with the mitochondrial marker cytochrome oxidase I

Institución donde se ejecutó el trabajo: Programa de Estudio y Control de Enfermedades Tropicales, PECET, Unidad de Malacología Médica y Trematodos (UMMT), Sede de Investigación Universitaria, Universidad de Antioquia, Medellín, Colombia Introducción. La fasciolosis es la enfermedad transmitida por vectores con mayor distribución latitudinal, longitudinal y altitudinal, debido a la capacidad colonizadora del parásito Fasciola hepatica y de sus huéspedes intermediarios, los moluscos limneidos. Estos caracoles se investigan por su importancia epidemiológica, pero su identificación taxonómica es difícil por la similitud fenotípica entre especies. En este sentido, con respecto a Lymnaea cousini , un huésped de F. hepatica en Colombia, existe incertidumbre en razón de su similitud morfológica con L. meridensis , descrita recientemente en Venezuela. Objetivo. Confirmar con el marcador del gen de la citocromo oxidasa I en el ADN mitocondrial COI (ADNmt), el estatus taxonómico de ejemplares morfológicamente caracterizados como L. cousini provenientes de Nariño, Norte de Santander y Santander (Colombia), depositados en la Colección de Moluscos Vectores de la Universidad de Antioquia, VHET N° 37. Materiales y métodos. Para la amplificación del COI mitocondrial, se extrajo ADN total del pie de cada ejemplar con el estuche DNeasy Blood and Tissue (Qiagen ® ). Los productos amplificados se enviaron a secuenciar a Macrogen Inc., Corea. Las 27 secuencias generadas en esta investigación se compararon con secuencias publicadas en el GenBank, incluidas las secuencias de la localidad tipo de L. cousini. Resultados. Se encontraron dos nuevos haplotipos de L. cousini para Colombia. Los especímenes de Nariño correspondían al haplotipo A, referenciado en Ecuador, y los especímenes de Santander y Norte de Santander, a un nuevo haplotipo al que se denominó D. Conclusión. Mediante el marcador mitocondrial del COI , se confirmó que los especímenes pertenecían a la especie L. cousini . Con el hallazgo se duplicó el número de haplotipos conocidos de la especie en Colombia y se amplió su distribución geográfica al suroeste y nordeste de la región altoandina colombiana.

Introduction: Fasciolosis is the disease transmitted by vectors with the highest latitudinal, longitudinal, and altitudinal distribution due to the colonizing capacity of the parasite Fasciola hepatica and its intermediate hosts, Lymnaeidae mollusks. These snails are under research due to their epidemiological importance, but their taxonomic identification is difficult given their interspecific phenotypical similarity. For this reason, there is uncertainty regarding Lymnaea cousini -a host of F. hepatica in Colombia- due to the morphological similarity it has with Lymnaea meridensis , recently described for Venezuela. Objective: To confirm with the COI marker (ADNmt) the taxonomic status of individuals morphologically identified as L. cousini from Nariño, Norte de Santander, and Santander (Colombia), deposited in the Vector Mollusks Collection VHET No. 37 of Universidad de Antioquia. Materials and methods: The amplification of the mitochondrial COI required total DNA extraction of each individual´s foot using the DNeasy Blood and Tissue Kit (Qiagen®). Products amplified were sent for sequencing to Macrogen Inc., Korea. Twenty seven sequences generated in this research were compared to sequences published in the GenBank, including sequences of the type locality of L. cousini . Results: Two new haplotypes of L. cousini were obtained for Colombia. Specimens from Nariño correspond to haplotype A, referenced for Ecuador, and specimens from Santander and Norte de Santander belong to a new haplotype we called haplotype D. Conclusion : By using the mitochondrial COI marker, we confirmed that the species under study did correspond to L. cousini . The number of known haplotypes of the species for Colombia has been duplicated and its geographical distribution has been extended to the southwest and northeast of the Colombian high Andean region.

EFFECTS OF SINGLE, BINARY AND TERTIARY COMBINATIONS WITH Jatropha gossypifolia AND OTHER PLANT-DERIVED MOLLUSCICIDES ON REPRODUCTION AND SURVIVAL OF THE SNAIL Lymnaea acuminata / Efeitos de combinações unitárias, binárias e terciárias de Jatropha gossypifolia e outros muluscicidas derivados de plantas na reprodução e sobrevivência do caramujo Lymnaea acuminata

The effect of sub-lethal doses (40% and 80% of LC50/24h) of plant derived molluscicides of singly, binary (1:1) and tertiary (1:1:1) combinations of the Rutin, Ellagic acid, Betulin and taraxerol with J. gossypifolia latex, leaf and stem bark powder extracts and their active component on the reproduction of freshwater snail Lymnaea acuminata have been studied. It was observed that the J. gossypifolia latex, stem bark, individual leaf and their combinations with other plant derived active molluscicidal components caused a significant reduction in fecundity, hatchability and survival of young snails. It is believed that sub-lethal exposure of these molluscicides on snail reproduction is a complex process involving more than one factor in reducing the reproductive capacity.

O efeito de doses sub-letais (40% e 80% de LC50/24h) de moluscicidas derivados de plantas com combinações unitárias, binárias (1:1) e terciárias (1:1:1) de Rutin, ácido Elágico, Betulin e taraxerol com látex da J. gossypifolia, folhas e extrato em pó de casca de caule e seus componentes ativos foram estudados na reprodução do caramujo de água fresca Lymnaea acuminata. Foi observado que o látex da J. gossypifolia, casca do caule, folhas individualmente e suas combinações com componentes moluscicidas ativos derivados de outras plantas causaram redução significante na fecundidade, incubação e sobrevivência dos caramujos jovens. Acredita-se que a exposição sub-letal destes moluscicidas sobre a reprodução dos caramujos é processo complexo envolvendo mais de um fator na redução da capacidade reprodutiva.

Distribution of bovine fasciolosis and associated factors in south Espírito Santo, Brazil: an update / Distribuição da fasciolose bovina e fatores associados no sul do Espírito Santo, Brasil: uma atualização

The geographical distribution and factors associated with bovine fasciolosis in the south of Espírito Santo were updated and the prevalences of this disease and of snails of the genus Lymnaea in the municipality of Jerônimo Monteiro were calculated. In the first stage, fecal samples were collected from 10% of the herds of 115 farms in 23 municipalities and interviews were conducted with owners. Generalized linear mixed models were used. In the second stage, in Jerônimo Monteiro municipality, feces and mollusks were collected from all farms registered in the milk cooperatives in the region. The mollusks were identified and examined for infection by Fasciola hepatica. Fasciolosis was diagnosed in 18 (78%) of the 23 municipalities. Of the 1157 fecal samples examined, 19.01% were positive for eggs of F. hepatica. The final model shows statistical evidence of associations between positive farms and previous cases of fasciolosis and concomitant grazing of cattle with other definitive hosts. In the evaluated farms from the studied municipality the prevalence of fasciolosis and Lymnaea was of 66.7% and 23.8%, respectively. Mollusks were found in flooded areas and the animals' drinking water troughs. The wide geographical distribution of bovine fasciolosis in the south of Espírito Santo requires control measures to prevent its expansion towards the north of this state and other places characterized as F. hepatica free-infection.

Realizou-se um estudo atualizado da distribuição geográfica e dos fatores associados à fasciolose bovina, no Sul do Estado do Espírito Santo, e calculadas a prevalência de moluscos do gênero Lymnaea no município de Jerônimo Monteiro. Na primeira etapa, coletaram-se amostras fecais de 10% dos rebanhos de 115 propriedades, em 23 municípios, e realizaram-se entrevistas com os proprietários. Modelos lineares generalizados mistos foram utilizados. Na segunda etapa, em Jerônimo Monteiro, coletaram-se fezes e moluscos em todas as propriedades cadastradas nas cooperativas de leite da região. Os moluscos foram identificados e examinados quanto à infecção por Fasciola hepatica. A fasciolose foi diagnosticada em 18 (78%) dos 23 municípios. Das 1157 amostras fecais examinadas, 19,01% foram positivas para ovos de F. hepatica. O modelo final mostra, concomitantemente, evidências estatísticas de associação entre propriedades positivas e casos anteriores de fasciolose e de pastoreio de bovinos com outros hospedeiros definitivos. Em Jerônimo Monteiro, a prevalência de fasciolose foi 66,7% e de Lymnaea 23,8%. Moluscos foram encontrados em áreas alagadas e em bebedouros dos animais. A ampla distribuição geográfica da fasciolose bovina, no Sul do Espírito Santo, requer medidas de controle que evitem sua expansão em direção ao Norte do Estado.

Distribution of bovine fasciolosis and associated factors in south Espírito Santo, Brazil: an update / Distribuição da fasciolose bovina e fatores associados no sul do Espírito Santo, Brasil: uma atualização

The geographical distribution and factors associated with bovine fasciolosis in the south of Espírito Santo were updated and the prevalences of this disease and of snails of the genus Lymnaea in the municipality of Jerônimo Monteiro were calculated. In the first stage, fecal samples were collected from 10% of the herds of 115 farms in 23 municipalities and interviews were conducted with owners. Generalized linear mixed models were used. In the second stage, in Jerônimo Monteiro municipality, feces and mollusks were collected from all farms registered in the milk cooperatives in the region. The mollusks were identified and examined for infection by Fasciola hepatica. Fasciolosis was diagnosed in 18 (78%) of the 23 municipalities. Of the 1157 fecal samples examined, 19.01% were positive for eggs of F. hepatica. The final model shows statistical evidence of associations between positive farms and previous cases of fasciolosis and concomitant grazing of cattle with other definitive hosts. In the evaluated farms from the studied municipality the prevalence of fasciolosis and Lymnaea was of 66.7% and 23.8%, respectively. Mollusks were found in flooded areas and the animals' drinking water troughs. The wide geographical distribution of bovine fasciolosis in the south of Espírito Santo requires control measures to prevent its expansion towards the north of this state and other places characterized as F. hepatica free-infection.

Realizou-se um estudo atualizado da distribuição geográfica e dos fatores associados à fasciolose bovina, no Sul do Estado do Espírito Santo, e calculadas a prevalência de moluscos do gênero Lymnaea no município de Jerônimo Monteiro. Na primeira etapa, coletaram-se amostras fecais de 10% dos rebanhos de 115 propriedades, em 23 municípios, e realizaram-se entrevistas com os proprietários. Modelos lineares generalizados mistos foram utilizados. Na segunda etapa, em Jerônimo Monteiro, coletaram-se fezes e moluscos em todas as propriedades cadastradas nas cooperativas de leite da região. Os moluscos foram identificados e examinados quanto à infecção por Fasciola hepatica. A fasciolose foi diagnosticada em 18 (78%) dos 23 municípios. Das 1157 amostras fecais examinadas, 19,01% foram positivas para ovos de F. hepatica. O modelo final mostra, concomitantemente, evidências estatísticas de associação entre propriedades positivas e casos anteriores de fasciolose e de pastoreio de bovinos com outros hospedeiros definitivos. Em Jerônimo Monteiro, a prevalência de fasciolose foi 66,7% e de Lymnaea 23,8%. Moluscos foram encontrados em áreas alagadas e em bebedouros dos animais. A ampla distribuição geográfica da fasciolose bovina, no Sul do Espírito Santo, requer medidas de controle que evitem sua expansão em direção ao Norte do Estado.

In vitro PHYTOTHERAPY OF VECTOR SNAILS BY BINARY COMBINATIONS OF LARVICIDAL ACTIVE COMPONENTS IN EFFECTIVE CONTROL OF FASCIOLIASIS / Fitoterapia in vitro de caramujos vetores por combinações binárias de componentes ativos larvicidas como controle efetivo de fasciolíase

SUMMARY A food-borne trematode infection fascioliasis is one among common public health problems worldwide. It caused a great economic loss for the human race. Control of snail population below a certain threshold level is one of the important methods in the campaign to reduce the incidence of fascioliasis. The life cycle of the parasite can be interrupted by killing the snail or Fasciola larva redia and cercaria inside of the snail Lymnaea acuminata. In vitro toxicity of different binary combinations (1:1 ratio) of plant-derived larvicidal active components such as citral, ferulic acid, umbelliferone, azadirachtin and allicin against Fasciola redia and cercaria were tested. The mortality of larvae was observed at 2h, 4h, 6h and 8h of treatment. In in vitro condition azadirachtin + allicin (1:1 ratio) was highly toxic against redia and cercaria (8h LC50 0.006 and 0.005 mg/L). Toxicity of citral + ferulic acid was lowest against redia and cercaria larvae. .

RESUMO A infecção alimentar pelo trematóide da fasciolíase é uma dentre os mais comuns problemas de saúde pública mundiais, causando grande prejuízo econômico para a humanidade. Controle da população de caramujos abaixo de determinado nível é um dos métodos no campo mais importantes para a redução da incidência da fasciolíase. O ciclo de vida do parasita pode ser interrompido pela morte do caramujo ou da larva redia e cercária da Fasciola dentro da Lymnaea acuminata. Foi testada a toxicidade in vitro das diferentes combinações binárias (relação 1:1) entre os vários componentes larvicidas ativos da planta tais como citral, ácido ferúlico, umbeliferone, azadiractina, e alicina contra a Fasciola redia e a cercária. A mortalidade das larvas foi observada após duas, quatro, seis e oito horas de tratamento. A condição in vitro azadiractina + alicina (relação 1:1) foi altamente tóxica contra redia e cercária (8h LC50 0,006 e 0,005 mg/L). Toxicidade do citral + ácido ferúlico foi a mais baixa contra redia e larvas de cercária. .

CHARACTERIZATION OF MOLLUSCICIDAL COMPONENT OF Moringa oleifera LEAF AND Momordica charantia FRUITS AND THEIR MODES OF ACTION IN SNAIL Lymnaea acuminata / Caracterização do componente moluscicida das folhas da Moringa oleifera e das frutas da Momordica charantia e seus modos de ação sobre o caramujo Lymnaea acuminata

SUMMARY The molluscicidal activity of the leaf powder of Moringa oleifera and lyophilized fruit powder of Momordica charantia against the snail Lymnaea acuminata was time and concentration dependent. M. oleifera leaf powder (96 h LC50: 197.59 ppm) was more toxic than M. charantia lyophilized fruit powder (96 h LC50: 318.29 ppm). The ethanolic extracts of M. oleifera leaf powder and Momordica charantia lyophilized fruit powder were more toxic than other organic solvent extracts. The 96 h LC50 of the column purified fraction of M. oleifera leaf powder was 22.52 ppm, while that of M. charantia lyophilized fruit powder was 6.21 ppm. Column, thin layer and high performance liquid chromatography analysis show that the active molluscicidal components in M. oleifera leaf powder and lyophilized fruit of M. charantia are benzylamine (96 h LC50: 2.3 ppm) and momordicine (96 h LC50: 1.2 ppm), respectively. Benzylamine and momordicine significantly inhibited, in vivo and in vitro, the acetylcholinesterase (AChE), acid and alkaline phosphatase (ACP/ALP) activities in the nervous tissues of L. acuminata. Inhibition of AChE, ACP and ALP activity in the nervous tissues of L. acuminata by benzylamine and momordicine may be responsible for the molluscicidal activity of M. oleifera and M. charantia fruits, respectively. .

RESUMO A atividade moluscicida do pó das folhas de Moringa oleifera e do pó liofilizado das frutas da Momordica charantia contra o caramujo Lymnaea acuminata é dependente do tempo e da sua concentração. O pó da folha da M. oleifera (96 h LC50: 197.59 ppm) foi mais tóxico do que o pó liofilizado da fruta da M. charantia (96 h LC50: 318.29 ppm). Os extratos etanólicos do pó de folha da M. oleifera e do pó liofilizado da fruta da M. charantia foram mais tóxicos do que outros extratos orgânicos solventes. O 96 h LC50 da fração purificada por coluna do pó das folhas da M. oleifera foi 22.52 ppm enquanto que o pó liofilizado do fruto da M. charantia foi 6.21 ppm. Coluna, camada fina e a alta performance da análise da cromatografia líquida mostram que os componentes ativos moluscicidas do pó da folha da M. oleifera e do liofiliizado da fruta da M. charantia são a benzilamina (96 h LC50: 22.3 ppm) e a momordicina (96 h LC50: 1.2 ppm), respectivamente. A benzilamina e a momordicina inibiram de maneira significante in vivo e in vitro a acetilcolinesterase (AChE), as atividades das fosfatases alcalina e ácida (ACP/ALP) nos tecidos nervosos da L. acuminata. A inibição da atividade da AChE, ACP e ALP nos tecidos nervosos da L. acuminata pela benzilamina e momordicina podem ser responsáveis pela atividade moluscicida da M. oleifera e dos frutos da M. charantia, respectivamente. .

Characterization of the molluscicidal activity of Bauhinia variegata and Mimusops elengi plant extracts against the fasciola vector lymnaea acuminata / Caracterização da atividade moluscicida dos extratos das plantas Bauhinia variegata e Mimusops elengi contra o vetor da Fasciola, Lymnaea acuminata

The molluscicidal activity of Bauhinia variegata leaf and Mimusops elengi bark was studied against vector snail Lymnaea acuminata. The toxicity of both plants was time and concentration-dependent. Among organic extracts, ethanol extracts of both plants were more toxic. Toxicity of B. variegata leaf ethanolic extract (96h LC50- 14.4 mg/L) was more pronounced than M. elengi bark ethanolic extract (96h LC50-15.0 mg/L). The 24h LC50 of column purified fraction of B. variegata and M. elengi bark were 20.3 mg/L and 18.3 mg/L, respectively. Saponin and quercetin were characterized and identified as active molluscicidal component. Co-migration of saponin (Rf 0.48) and quercetin (Rf 0.52) with column purified bark of M. elengi and leaf of B. variegata on thin layer chromatography demonstrate same Rf value i.e. 0.48 and 0.52, respectively. The present study clearly indicates the possibility of using M. elengi and/or B. variegata as potent molluscicide.

A atividade moluscicida das folhas da Bauhinia variegata e da casca do Mimusops elengi foi testada contra o vetor caracol, Limnaea acuminata. A toxicidade de ambas as plantas é dependente do tempo e da concentração. Entre os extratos orgânicos, os extratos de etanol de ambas as plantas foi mais tóxico. A toxicidade do extrato etanólico da folha da B. variegata (96 h LC50 - 14,4 mg/L) foi mais pronunciada do que o extrato etanólico da casca do M. elengi (96h - LC50 - 15,0 mg/L). As frações purificadas em coluna durante 24 h LC50 do B. variegata e da casca do M. elengi foram 20,3 mg/L e 18,3 mg/L, respectivamente. A saponina e a quercentina foram caracterizadas e identificadas como os componentes ativos moluscicidas. A co-migração da saponina (Rf 0,48) e da quercentina (Rf 0,52) com a casca purificada por coluna do M. elengi e as folhas da B. variegata na cromatografia demonstraram o mesmo valor Rf isto é, 0,48 e 0,52 respectivamente. O presente estudo indica claramente a possibilidade de usar M. elengi e/ou B. variegata como moluscicidas potentes.

DNA multigene sequencing of topotypic specimens of the fascioliasis vector Lymnaea diaphana and phylogenetic analysis of the genus Pectinidens (Gastropoda)

Freshwater lymnaeid snails are crucial in defining transmission and epidemiology of fascioliasis. In South America, human endemic areas are related to high altitudes in Andean regions. The species Lymnaea diaphana has, however, been involved in low altitude areas of Chile, Argentina and Peru where human infection also occurs. Complete nuclear ribosomal DNA 18S, internal transcribed spacer (ITS)-2 and ITS-1 and fragments of mitochondrial DNA 16S and cytochrome c oxidase (cox)1 genes of L. diaphana specimens from its type locality offered 1,848, 495, 520, 424 and 672 bp long sequences. Comparisons with New and Old World Galba/Fossaria, Palaearctic stagnicolines, Nearctic stagnicolines, Old World Radix and Pseudosuccinea allowed to conclude that (i) L. diaphana shows sequences very different from all other lymnaeids, (ii) each marker allows its differentiation, except cox1 amino acid sequence, and (iii) L. diaphana is not a fossarine lymnaeid, but rather an archaic relict form derived from the oldest North American stagnicoline ancestors. Phylogeny and large genetic distances support the genus Pectinidens as the first stagnicoline representative in the southern hemisphere, including colonization of extreme world regions, as most southern Patagonia, long time ago. The phylogenetic link of L. diaphana with the stagnicoline group may give light to the aforementioned peculiar low altitude epidemiological scenario of fascioliasis.

Impact of Taraxerol in combination with extract of Euphorbia tirucalli plant on biological parameters of Lymnaea acuminata / Impacto do Taraxerol em combinação com o extrato da planta de Euphorbia tirucalli nos parâmetros biológicos da Lymnaea acuminata

The present work was carried out to evaluate the molluscicidal activity of active ingredient Taraxerol with the acetone extract of Euphorbia tirucalli against Lymnaea acuminata snail. The (fecundity, hatchability and survivability) of snail L. acuminata exposed to this extract was studied. The effects of the tested extracts on life-history traits of harmful snail L. acuminata have also been evaluated, and this study also expounds the inhibitory effects of these extracts singly as well as in binary combination (1:1 ratio). It concluded that these herbal products act as a potential source of molluscicides, and that they would also have the advantage of easy availability, low cost, biodegradability and greater acceptance amongst users than synthetic pesticide.

O presente trabalho foi levado a efeito para avaliar a atividade moluscicida do ingrediente ativo Taraxerol junto com o extrato de acetona da Euphorbia tirucalli contra o caramujo Lymnaea acuminata. Foram estudados a fecundidade, o desenvolvimento e sobrevivência do caramujo Lymnaea acuminata exposto a estes extratos. Os efeitos danosos destes extratos testados na vida do caramujo L. acuminata foram também avaliados e este estudo demonstra os efeitos inibidores destes extratos isolados ou em combinação binária (relação l:1). Concluiu-se que estes produtos vegetais agem como fontes potenciais de moluscicidas e que eles também têm a vantagem de obtenção fácil, baixo custo, biodegradabilidade e maior aceitação entre os usuários do que os pesticidas sintéticos.

Bait formulations of molluscicides and their effects on biochemical changes in the ovotestis of snail Lymnaea acuminata (Mollusca; Gastropoda:Lymnaeidae) / Formulações de iscas de moluscicidas e seus efeitos sobre as alterações bioquímicas no ovoteste do caramujo Lymnaea acuminata (Mollusca;Gastropoda:Lymnaeidae)

The effect of sub-lethal feeding of bait formulations containing molluscicidal component of Ferula asafoetida (ferulic acid, umbelliferone), Syzygium aromaticum (eugenol) and Carum carvi (limonene) on biochemical changes in the ovotestis of snail Lymnaea acuminata were studied. Bait formulations feeding to L. acuminata were studied in clear glass aquaria having diameter of 30 cm. Baits were prepared from different binary combinations of attractant amino acid (valine, aspartic acid, lysine and alanine 10 mM) in 100 mL of 2 percent agar solution + sub-lethal (20 percent and 60 percent of 24h LC50) doses of different molluscicides (ferulic acid, umbelliferone, eugenol and limonene). These baits caused maximum significant reduction in free amino acid, protein, DNA, RNA levels i.e. 41.37, 23.56, 48.36 and 14.29 percent of control in the ovotestis of the snail, respectively. Discontinuation of feeding after treatment of 60 percent of 96h LC50 of molluscicide containing bait for next 72h caused a significant recovery in free amino acid, protein, DNA and RNA levels in the ovotestis of L. acuminata.

Foi estudado o efeito subletal das iscas usadas para alimentação contendo componentes moluscicidas de Ferula asafoetida (ácido ferúlico, umbeliferone), Syzygium aromaticum (eugenol) e Carum carvi (limonene) nas alterações bioquímicas do ovoteste do caramujo Lymnaea acuminata. A formulação das iscas usadas para alimentar L. acuminata foi estudada em aquários de vidros transparentes de diâmetro de 30 cm. As iscas foram preparadas por combinações diferentes binárias de aminoácidos (valina, ácido aspártico, lisina e alanina 10 mM) em 100 mL de solução de agar a 2 por cento + doses subletais (20 por cento e 60 por cento durante 24 horas LC50) de diferentes moluscicidas (ácido ferúlico, umbeliferone, eugenol e limonene). Estas iscas causaram redução significante máxima em aminoácidos livres, proteínas, níveis de DNA e RNA isto é 41,37 por cento, 23,56 por cento, 48,36 por cento e 14,29 por cento de controle no ovoteste do caramujo, respectivamente. Discontinuação da alimentação depois do tratamento de 60 por cento de 96 horas de LC50 do moluscicida contendo a isca para as subsequentes 72 horas causou significante recuperação dos níveis de aminoácidos livres, proteína, DNA e RNA no ovoteste da L. acuminata.

Efficacy of euphorbia hirta latex as plant derived molluscicides against freshwater snails / Eficácia do látex da Euphorbia hirta como moluscicida vegetal contra caramujos de água doce

The toxic effect of binary and tertiary combinations of Euphorbia hirta Linn latex powder with other plant molluscicidal compounds, were evaluated against the freshwater snails Lymnaea (Radix) acuminata and Indoplanorbis exustus in pond. These combinations showed significant time and dose dependent effect against both the snails. These compounds at higher doses were also lethal to freshwater fish Channa punctatus (Bloch) (Channidae {Ophicephalidae}), which shares the habitat with these snails, but the LC90 (24h) doses of snails have no apparent killing properties in fish populations when treated in mixed population of snails and fish.

Os efeitos tóxicos das combinações binárias e terciárias do pó de látex da Euphorbia hirta Linn assim como outros compostos vegetais moluscicidas foram avaliados em sua ação sobre caramujos de água doce Lymnaea (Radix) acuminata e Indoplanorbis exustus em represas. Estas combinações mostraram significante efeito dose e tempo dependente contra ambos os caramujos. Estes compostos em doses altas foram também letais para peixes de água doce Channa punctatus (Bloch) (Channidae {Ophicephalidae}), que compartilham o ambiente com estes caramujos mas a dose LC90 (24h) para caramujos aparentemente não tem propriedade de matar as populações de peixes quando uma população mista de peixes e caramujos são tratadas.

Descripción morfológica de Cotylophoroncotylophorum (Digenea: Paramphistomidae) hallado enbovinos de Rionegro, Antioquia, Colombia / Descrição morfológica do Cotylophoron cotylophorum (Digenea: Paramphistomidae)em bovinos de Rionegro, Antioquia, Colômbia / Morphological description of Cotylophoron cotylophorum (Digenea: Paramphistomidae) in Cattle from Rionegro, Antioquia, Colombia

En Colombia se ha registrado la presencia de Paramphistomidae (Trematoda: Digenea) parasitandoel rumen de bovinos de la costa Caribe, los Llanos Orientales y el suroccidente. En estas publicacionesse menciona la presencia y prevalencia de los paramfistómidos, pero todas carecen de la descripciónmorfológica de los digeneos. El hallazgo de paramfistómidos adultos en el rumen de ejemplaresBos taurus, procedentes del sector Llanogrande, en el municipio de Rionegro, Antioquia, motivó lapresente investigación, con el propósito de describir los diferentes estadíos del parásito y establecerel estatus taxonómico específico. Del rumen de una hembra, Bos taurus, sacrificada en la CentralGanadera del municipio de Medellín, se extrajeron vivos 52 digeneos adultos. En el laboratoriose aislaron huevos que se incubaron hasta la obtención de miracidios. Con ellos se infectaron 50moluscos Lymnaea truncatula. La morfología de los adultos y estadíos larvarios del paramfistómidopermitió determinarlo como Cotylophoron cotylophorum. En L. truncatula se obtuvo una infeccióndel 5.1 %. Con este registro se amplia la distribución geográfica y altitudinal del parásito a la regiónaltoandina, noroccidental de Colombia.

The presence of Paramphistomidae parasites (Trematoda: Digenea) inside the rumen (first stomach) hasbeen recorded in Colombia in cattle from the Caribbean coast, the Eastern Plains region in the east, and thesouthwest of the country. However, studies on the presence and prevalence of the parasite do not include anymorphological description. This study includes research findings from adult Paramphistomidae parasitesin the rumen of Bos taurus specimens from Llanogrande in the municipality of Rionegro, Antioquia. Theaim of this research was to describe all the developmental stages of the parasite and to establish its specifictaxonomical status. 52 live adult parasites were found in the rumen of a female Bos taurus sacrificed inthe slaughter house Central Ganadera of Medellin. Eggs from adults were obtained in the laboratory, andhatching was stimulated to obtain miracidia. 50 Lymnaea truncatula snails were exposed to the miracidia,resulting in an infection rate of 5.1 %. The parasite was identified as Cotylophoron cotylophorum byusing adult morphology and larval stages. This finding expands the parasite’s geographical and altitudinaldistribution to the andean highlands in northwestern Colo.

Na Colômbia tem-se encontrado a presencia de Paramphistomidae (Trematoda: Digenea) parasitandono rúmen de bovinos da costa Caribe, os Llanos Orientales e no sul oeste. Nestas publicações tem-semencionado a presencia e prevalência dos paramfistómidos, mas todas carecem da descrição morfológicados digeneos. Do rúmen de uma fêmea Bos taurus proveniente do município de Rionegro e sacrificadana Central Ganadera do município de Medellín, se extraíram vivos 52 digeneos adultos. No laboratórioforam isolados os ovo e forma obtidos os miracidios. Com eles foram infetados 50 moluscos Lymnaeatruncatula. A morfologia dos adultos e estádios larvários do paramfistómido permitiu determinar-lho comoCotylophoron cotylophorum. Em L. truncatula foi obtido uma infecção de 5.1 %. Com este fato, se ampliaa distribuição geográfica e altitudinal do parasita na região alta dos Andes colombianos.

Occurrence of Fasciola hepatica (Linnaeus, 1758) infection in Brazilian cattle of Minas Gerais, Brazil / Ocorrência de Fasciola hepatica Linnaeus, 1758 em rebanhos bovinos do estado de Minas Gerais, Brasil

Fasciolosis is a parasitic disease which is caused by digenetic trematodes, such as Fasciola hepatica, and which occurs worldwide. The disease causes significant economic losses in cattle because of the reduction in milk and meat production and because the parasitized livers have no economic value. Also, abortion and mortality rates increase. To determine the occurrence and dispersal of F. hepatica in the state of Minas Gerais, fecal samples were collected from bovines aged 12 months and older between April 2005 and April 2006. The bovines came from 1,251 rural farms of 120 municipalities/cities in Minas Gerais. Water was collected to ascertain the presence of the intermediate host in the properties where infected bovines were found. A total of 16 municipalities had infected bovines and 13 had the presence of Lymnaea columella. We detected infected bovines in 70 percent of the rural farms visited in the municipality of Itajubá. Itajubá was also found to have infected snails with the larval stage of F. hepatica. The results revealed that infection by F. hepatica in bovines of Minas Gerais was associated with the presence of the intermediate host and favorable epidemiological factors, drawing attention to the necessity of adopting proper control measures in the area.

A fasciolose é uma doença parasitária que ocorre em todo o mundo e é causada por um trematódeo digenético conhecido como Fasciola hepatica. A doença é responsável por causar perdas econômicas significativas em bovinos, devido à diminuição na produção de carne e leite, condenação de fígados parasitados e aumento na mortalidade. Para determinar a ocorrência e a dispersão da F. hepatica no estado de Minas Gerais, amostras de fezes de bovinos com idade superior a 12 meses foram coletadas, no período de Abril de 2005 a Abril de 2006. Foram examinados bovinos provenientes de 1.251 propriedades rurais de 120 municípios do estado de Minas Gerais. Coleções hídricas também foram examinadas, a fim de verificar a presença de hospedeiros intermediários nas propriedades onde foram encontrados bovinos parasitados. Para um total de 16 municípios foi relatada a presença de bovinos infectados por F. hepatica e destes, 13 apresentaram também o molusco Lymnaea columella. Foi possível diagnosticar bovinos infectados em 70 por cento das propriedades visitadas no município de Itajubá. Além disso, no município de Itajubá foi possível detectar a presença de moluscos naturalmente infectados com diferentes estádios larvais de F. hepatica. Os resultados evidenciam a necessidade de medidas de controle para esta parasitose na área estudada, uma vez que foi possível verificar a presença de bovinos e moluscos infectados por F. hepatica, associada a fatores epidemiológicos favoráveis a dispersão do parasito.

Occurrence of Fasciola hepatica (Linnaeus, 1758) infection in Brazilian cattle of Minas Gerais, Brazil

Fasciolosis is a parasitic disease which is caused by digenetic trematodes, such as Fasciola hepatica, and which occurs worldwide. The disease causes significant economic losses in cattle because of the reduction in milk and meat production and because the parasitized livers have no economic value. Also, abortion and mortality rates increase. To determine the occurrence and dispersal of F. hepatica in the state of Minas Gerais, fecal samples were collected from bovines aged 12 months and older between April 2005 and April 2006. The bovines came from 1,251 rural farms of 120 municipalities/cities in Minas Gerais. Water was collected to ascertain the presence of the intermediate host in the properties where infected bovines were found. A total of 16 municipalities had infected bovines and 13 had the presence of Lymnaea columella. We detected infected bovines in 70% of the rural farms visited in the municipality of Itajubá. Itajubá was also found to have infected snails with the larval stage of F. hepatica. The results revealed that infection by F. hepatica in bovines of Minas Gerais was associated with the presence of the intermediate host and favorable epidemiological factors, drawing attention to the necessity of adopting proper control measures in the area.

A fasciolose é uma doença parasitária que ocorre em todo o mundo e é causada por um trematódeo digenético conhecido como Fasciola hepatica. A doença é responsável por causar perdas econômicas significativas em bovinos, devido à diminuição na produção de carne e leite, condenação de fígados parasitados e aumento na mortalidade. Para determinar a ocorrência e a dispersão da F. hepatica no estado de Minas Gerais, amostras de fezes de bovinos com idade superior a 12 meses foram coletadas, no período de Abril de 2005 a Abril de 2006. Foram examinados bovinos provenientes de 1.251 propriedades rurais de 120 municípios do estado de Minas Gerais. Coleções hídricas também foram examinadas, a fim de verificar a presença de hospedeiros intermediários nas propriedades onde foram encontrados bovinos parasitados. Para um total de 16 municípios foi relatada a presença de bovinos infectados por F. hepatica e destes, 13 apresentaram também o molusco Lymnaea columella. Foi possível diagnosticar bovinos infectados em 70% das propriedades visitadas no município de Itajubá. Além disso, no município de Itajubá foi possível detectar a presença de moluscos naturalmente infectados com diferentes estádios larvais de F. hepatica. Os resultados evidenciam a necessidade de medidas de controle para esta parasitose na área estudada, uma vez que foi possível verificar a presença de bovinos e moluscos infectados por F. hepatica, associada a fatores epidemiológicos favoráveis a dispersão do parasito.