Detection and genetic distance of resistant populations of Pseudosuccinea columella (Mollusca: Lymnaeidae) to Fasciola hepatica (Trematoda: Digenea) using RAPD markers.

Twelve natural populations of Pseudosuccinea columella snails, sampled in the western and central regions of Cuba, were analyzed using the RAPD-PCR technique to screen for resistance to Fasciola hepatica. Ten OPA primers previously shown to produce marker bands for resistance and susceptibility were tested. A new population of P. columella (El Azufre, Pinar del Río) exhibited the amplification patterns of resistant snails, and its resistant status was confirmed after experimental exposure to miracidia. No genetic variability was detected across or within the susceptible isolates. Similarly, the novel resistant isolate displayed an RAPD profile identical to the profile of two other isolates previously identified as resistant to F. hepatica. However, clear differences in RAPD banding patterns and genetic distance were observed between resistant and susceptible isolates.

Population dynamics of intermediate snail hosts of Fasciola hepatica and some environmental factors in San Juan y Martinez municipality, Cuba.

The variation of abundances of intermediate snail hosts of Fasciola hepatica in Cuba (Fossaria cubensis and Pseudosuccinea columella) was studied during one year under natural conditions at five sampling sites in San Juan y Martinez municipality, Pinar del Rio province, Cuba. The effect of some environmental variables on the lymnaeid abundances was also studied. A canonical correspondence analysis showed that both species do not generally occur together in the same habitat and that most factors affect them in an opposite fashion, although both of them correlate positively through time to the diversity of the habitats. F. cubensis prefers the sites that are in or closer to the city whereas P. columella is more abundant in rural sites. Lymnaeid abundances are mainly affected by nitrite and nitrate concentrations as well as by the abundance of the thiarid Tarebia granifera. F. cubensis is more abundant in polluted habitats with low densities (or absence) of T. granifera whereas P. columella prefers cleaner habitats and can coexist with the thiarid, even at its higher densities. The implications of divergent preferences of the two lymnaeids for the control of fasciolosis are discussed.

Population dynamics of intermediate snail hosts of Fasciola hepatica and some environmental factors in San Juan y Martinez municipality, Cuba

The variation of abundances of intermediate snail hosts of Fasciola hepatica in Cuba (Fossaria cubensis and Pseudosuccinea columella) was studied during one year under natural conditions at five sampling sites in San Juan y Martinez municipality, Pinar del Rio province, Cuba. The effect of some environmental variables on the lymnaeid abundances was also studied. A canonical correspondence analysis showed that both species do not generally occur together in the same habitat and that most factors affect them in an opposite fashion, although both of them correlate positively through time to the diversity of the habitats. F. cubensis prefers the sites that are in or closer to the city whereas P. columella is more abundant in rural sites. Lymnaeid abundances are mainly affected by nitrite and nitrate concentrations as well as by the abundance of the thiarid Tarebia granifera. F. cubensis is more abundant in polluted habitats with low densities (or absence) of T. granifera whereas P. columella prefers cleaner habitats and can coexist with the thiarid, even at its higher densities. The implications of divergent preferences of the two lymnaeids for the control of fasciolosis are discussed.

Evidence of phenotypic differences between resistant and susceptible isolates of Pseudosuccinea columella (Gastropoda: Lymnaeidae) to Fasciola hepatica (Trematoda: Digenea) in Cuba.

Isolates of Cuban Pseudosuccinea columella susceptible and non-susceptible to Fasciola hepatica miracidial infection were compared for cellular reaction to miracidial development, shell morphometrics, mantle pigmentation pattern, and egg laying behaviour. At 24 h post-exposure, non-susceptible snails showed an early cellular host reaction to encapsulate and phagocytise the miracidium, whereas viable transforming miracidia were observed in exposed susceptible snails, indicating that non-susceptibility resulted from resistance rather than physiological unsuitability. Susceptible isolates (more than 80% susceptible to infection) showed significantly higher values of the morphometric indices shell width/length and aperture width/length than resistant isolates (100% resistant). Mantle pigmentation pattern was analysed using Visilog software; the cumulative distances between mantle spots were plotted and compared between isolates, showing a clear distinction between groups. Egg-laying behaviour also showed significant differences with susceptible snails preferring to lay their eggs on the container's walls whereas resistant snails preferred to lay their eggs on the bottom of the container.

Fasciola hepatica: identification of molecular markers for resistant and susceptible Pseudosuccinea columella snail hosts.

Protein electrophoresis, RAPD-PCR and nuclear rDNA ITS sequencing were performed to search for genetic differences between Pseudosuccinea columella snails susceptible and resistant to Fasciola hepatica infection. Of the 21 enzymatic loci analyzed in both populations, none of them exhibited neither within- or between-group variation. Such an absence of enzyme polymorphism support the hypothesis of selfing as the "prevalent" mating system for this hermaphroditic species. Conversely, the RAPD profiles displayed clear differences between susceptible and resistant isolates for 17 of the 26 primers tested while no within-group variation was detected. rDNA ITS sequence analysis from snails of each isolates showed only two bases that differed between groups accounting for a 0.17% of variation confirming that susceptible and resistant snails belong to the same species. This is the first time that a genetic variation using RAPD markers is demonstrated between susceptible and resistant lymnaeid snails vis-a-vis of F. hepatica infection in absence of experimental selection.

The effect of isolation on reproduction and growth of Pseudosuccinea columella (Pulmonata: Lymnaeidae): a snail-conditioned water experiment.

A snail-conditioned water experiment was conducted in Pseudosuccinea columella to test the possible role of a chemical interaction between snails on the diminished growth and fecundity rates found for snails raised in pairs compared to those raised in complete isolation. The results permit to discard the hypothesis of an inhibition of growth and reproduction between snails due to factors released into the water.

The effect of isolation on reproduction and growth of Pseudosuccinea columella (Pulmonata: Lymnaeidae): a snail-conditioned water experiment

A snail-conditioned water experiment was conducted in Pseudosuccinea columella to test the possible role of a chemical interaction between snails on the diminished growth and fecundity rates found for snails raised in pairs compared to those raised in complete isolation. The results permit to discard the hypothesis of an inhibition of growth and reproduction between snails due to factors released into the water

Fasciola hepatica (Trematoda: Digenea): its effect on the life history traits of Pseudosuccinea columella (Gasteropoda: Lymnaeidae), an uncommon interaction.

An uncommon snail-trematode interaction pattern was found for the Pseudosuccinea columella - Fasciola hepaticamodel under laboratory conditions. Week-old juveniles from two isolates showed very high rates of infection (90.0% and 93.3%) after exposure to five miracidia. They also showed differences in their life history traits when compared to a control group. However, they did not exhibit the increase in size and reduction/cessation in host fecundity which is usually observed in most of the other related snail-trematode systems. In contrast, infected juvenile P. columella showed increased egg laying after the onset of cercarial emission and there was no effect on growth. A third isolate of P. columella was found to be refractory to miracidial infection. This constitutes the first report of non-susceptibility to F. hepatica in a lymnaeid species which is usually susceptible. These non-susceptible snails exhibited lower fecundity and survival compared to non-exposed susceptible snails under identical laboratory conditions. The differences observed in terms of life span and reproduction could result from the cost of resistance for individuals from this isolate.

The effect of isolation on the life-history traits of Pseudosuccinea columella (Pulmonata: Lymnaeidae)

A population of Pseudosuccinea columella was raised under laboratory conditions and its life tables were determined in isolated and paired snails. Isolated snails were significantly larger in shell size than paired snails from five weeks of age onward. Also, statistically significant differences were found for the number of eggs per mass per individual from week 5 to 9, isolated snails exhibiting the highest values. The intrinsic and finite rates of increase were greater in isolated than in paired snails. Either an inhibition of the reproductive output between individuals or the advantage of selfing may be the cause of the differences in this species, acting as a possible mechanism that increase the fitness of isolated snails

Life-history traits of Fossaria cubensis (Gastropoda: Lymnaeidae) under experimental exposure to Fasciola hepatica (Trematoda: Digenea)

The effect of exposing the lymnaeid snail Fossaria cubensis to the trematode Fasciola hepatica on the snail population's life-history traits was studied under laboratory conditions. Exposed individuals showed a lower survival rate than control snails, although from week 7 onward a slower decrease of this parameter in relation to the control group was observed. There were higher values of fecundity rate for the controls compared to the exposed group except during weeks 9, 10, 11 and 12, which was the time that followed the period when almost all of the infected snails died. Both the intrinsic and finite rates of natural increase were significantly higher for the control group, but exposed snails still attained a lower mean generation time. Age-specific trade-offs were found, mainly for the weekly increase in size versus the number of eggs per mass, the weekly increase in size versus the number of viable eggs per mass, the number of masses versus the hatching probability and the number of eggs versus the hatching probability. All these negative associations were significant for juveniles of both control and exposed snails and not for adults; however, exposed young individuals exhibited much higher values of the correlation coefficient than control animals.

Evaluación de la acción molusquicida de agave legrelliana sobre fossaria cubensis (mollusca: lymnaeidae), principal vector de fascioliasis en Cuba / Evaluation of the molluscicidal action of agave legrelliana on fossaria cubensis (mollusca: lymnaeidae) main vector of fascioliasis in Cuba

Se determinaron las dosis letales LD 50 y LD 90 del jugo extraído de las hojas de Agae legrelliana (Agavacea) sobre Fossaria cubensis, en condiciones de laboratorio, alcanzando los valores de 0,105 y 7.178 ml/l respectivamente. Aplicando estos valores se comprobó que éstas provocan una disminución notable en la frecuencia cardíaca de los moluscos

Comparación del crecimiento de Helisoma duryi (Wetherby, 1879); Helisoma scalare (Jay, 1839), (Mollusca: Planorbidae) / Comparison betew the growth of Helisoma duryi (Wetherby, 1879); Helisoma scalare (Jay, 1893), Mollusca: Planorbidae)

Se hace una comparación del crecimiento de Helisoma duryi (Wetherby, 1879) y Helisoma scalare (Jay, 1839) en condiciones naturales. Se informa que ambas especies se mantuvieron en cajas de cría en la Ciénaga de Zapata y se les calculó la curva de crecimiento mediante la ecuación de von Bertalanffy. Se indica que el crecimiento de estas especies en cuanto al diámetro máximo alcanzable es similar, pero los valores de las constantes de crecimiento señalan que H. duryi crece más rápidamente que H. scalare. Se plantea que este hecho puede contribuir a una ubicación sistemática definitiva de estas especies

Estudio de los moluscos fluviátiles de una localidad afectada por un brote de fascioliasis / Fluvial mollusks from a locality affected by a fascioliasis outbreak

Se informa que se ralizó un estudio de las diferentes especies de moluscos dulceacuícolas en la localidad de Fomento, afectada por un brote de fascioliasis. Se plantea que en el área del municipio se colectaron diferentes especies de moluscos, encontrándose entre éstos Lymnaea cubensis (Pfeiffer, 1839) y Lymnaea columella (Say, 1871): hospederos intermediarios de Fasciola hepatica (Linne, 1758). Se expone que los análisis de laboratorio corroboaron que L. cubensis es el principal vector en esta epidemia. Se indica la necesidad de realizar los estudios pertinentes sobre la posibilidad de aplicar el método de control biológico a los vectores de esta enfermedad