Seasonal dynamics of Fasciola hepatica burdens in grazing Timahdit sheep in Morocco

Seasonal transmission of Fasciola hepatica was observed in sentinel sheep and the dynamics of the snail intermediate host, Lymnaea truncatula, was followed over a 3-year study period in the Middle-Atlas mountains in Morocco. High fluke burdens were recorded in both lambs and ewes in the fall and winter, suggesting that transmission occurred in late spring. Fluke burdens ranged from one to 302 in ewes and from one to 345 in lambs. Infections with 200 or more flukes were always fatal. A unique feature of this study was the annual cyclical fluctuation of the fluke burdens. Burdens reached maximum levels during the winter and then declined to low numbers by late spring and summer. This suggested self-regulation which may be dependent on breed resistance or may be related to forage factors, including lack of forage (nutritional stress). Snail populations were cyclical and correlated with fluke transmission as observed in the sentinel sheep. The weather was observed to affect the snail populations which in turn limited fluke transmission.     

Lack of molluscan host diversity and the transmission of an emerging parasitic disease in Bolivia

Fasciolosis is a re-emerging parasitic disease that affects an increasing number of people in developing countries. The most severe endemic affects the Bolivian Altiplano, where the liver fluke (Fasciola hepatica) and its hermaphroditic snail host, Lymnaea truncatula, have been introduced from Europe. To achieve a better understanding of the epidemiological situation and the consequences of the colonization event of this invasive species, genetic analysis of Bolivian snail populations was needed. Here we compare the genetic diversity and population structure of snail samples from the Bolivian Altiplano with samples from the Old World at six polymorphic microsatellite loci. Whereas some variability exists in the snail populations from the Old World, we observe only a single genotype of L. truncatula in the Bolivian Altiplano. We discuss the possible explanations for such a reduction in genetic variability, and, given the high natural parasitism pressures exerted on the snail populations, we discuss the relevance of this result for host-parasite interactions.     

Population dynamics of Lymnaea columella and its natural infection by Fasciola hepatica in the State of Minas Gerais,Brazil

Studies on the population dynamics and natural infection of Lymnaea columella by Fasciola hepatica were carried out from September 1999 to December 2000 in a low-lying area near Itajubá in the Brazilian State of Minas Gerais. A total of 626 snails were collected monthly at nine different sites, counted, and dissected to search for larvae of F. hepatica. The highest populations of L. columella were reached in October of 1999 and August of 2000, and the highest natural infection rates of snails by F. hepatica were reached in September 1999 (5.2%) and July 2000 (3.9%). The removal by farmers of aquatic plants from the drainage furrows caused a drastic reduction in this snail population.     

Southernmost finding of Lymnaea viatrix orbigny, 1835 (Pulmonata: Lymnaeidae), intermediate host of Fasciola hepatica (Linnaeus, 1758) (Trematoda: Digenea), in urban and rural areas of Patagonia, Argentina

We report the first finding of Lymnaea viatrix south of parallel 41 masculine S, in rural and urban areas from Argentina. Ninety snails were collected during year 2000, from a concrete pond at a Public Square in El Bolsón Village, Río Negro province, and 811 snails in November 1999, and during 2000 from waterbodies within a farm at Cholila locality, Chubut province. Fasciola hepatica infection was detected in 0.9% snails from the rural area. We discuss the potential risk of L. viatrix to public health in urban areas and its epidemiological importance in rural areas of the Andean Patagonian region.     

The susceptibility of Lymnaea fuscus to experimental infection with Fasciola hepatica

Three experiments on the infection of Lymnaea fuscus with Fasciola hepatica were carried out to determine if successful infections and maturation of the parasite were dependent on the size of snails at miracidial exposure. The first experiment was performed using 1-4-mm-high snails from 2 populations of L. fuscus and 1 population of Lymnaea palustris. In these snails each subjected to a single bimiracidial exposure, the prevalence of F. hepatica infection at day 35 postexposure ranged from 20.3% to 46.2% in snails measuring 1 mm in height at exposure; it was lower in the 2-mm snails and was 0 in higher size classes. The second experiment was performed by subjecting 1- and 4-mm L. fuscus to 1, 2, and 3 bimiracidial exposures. The prevalence of F. hepatica infection at day 35 postexposure was maximum in the 1-mm snails exposed once to miracidia and decreased with increasing number of exposures. The results were negative in 4-mm snails. Cercarial shedding of F. hepatica was studied in the third experiment using 1- and 2-mm L. fuscus each subjected to a single bimiracidial exposure. The total number of cercariae released from these snails was less than 50. From these results, it can be concluded that L. fuscus showed a partial resistance to F. hepatica infection due to snail age.     

First report of Lymnaea columella Say, 1817 (Pulmonata: Lymnaeidae) naturally infected with Fasciola hepatica (Linnaeus,1758) (Trematoda: Digenea) in Argentina

We report the first evidence of natural infection of Lymnaea columella with Fasciola hepatica in Argentina. A sample of 601 snails was collected in May 2003 in northeastern Corrientes, a province bounded on the north by Paraguay, on the east by Brazil and on the southeast by Uruguay. Among 500 examined snails, 44 (8.8%) were exclusively infected with F. hepatica. Parasite identification was based on morphological features of cercariae from snails, and of eggs and adult flukes from Wistar rats. We discuss the events suggesting that an enzootic transmission cycle of F. hepatica has been recently established in northeastern Corrientes.     

Aspects of the maintenance of the life cycle of Fasciola hepatica in Lymnaea columella in Minas Gerais,Brazil

Fascioliasis is a parasitic disease of domestic ruminants that occurs worldwide. The lymnaeid intermediate hosts of Fasciola hepatica include Lymnaea columella, which is widely distributed in Brazil. A colony of L. columella from Belo Horizonte, MG, was reared in our laboratory to be used in studies of the F. hepatica life cycle, the intermediate host-parasite relationship and development of an anti-helminthic vaccine. In the first experiment 1,180 snails were exposed to miracidia of F. hepatica eggs removed from the biliary tracts of cattle from the State of Rio Grande do Sul. In the second and third experiments the snails were exposed to miracidia that had emerged from F. hepatica eggs from Uruguay, maintained in rabbits. The rates of infection in the first, second and third experiments were 0, 42.1 and 0% respectively. Over 15,806 metacercariae were obtained and stored at 4 degrees C. Four rabbits weighing 1.5 kg each were infected with 32-44 metacercariae and two with 200. Three rabbits begin to eliminate eggs of the parasite in the feces from 84 days after infection onwards. The biological cycle of F. hepatica in L. columella and the rabbit was completed within 124 days.     

Characterization of sheep antibodies involved in precipitate formation with surface antigens of Fasciola hepatica in vitro

Sandeman R. M. and Howell M. J. 1982. Characterization of sheep antibodies involved in precipitate formation with surface antigens of Fasciola hepatica in vitro. International Journal for Parasitology12: 467–471. The role of sheep antibodies which precipitate with surface antigens of Fasciola hepatica is unclear. In an attempt to clarify their function these antibodies were characterized as to their immunoglobulin class and ability to affect the survival of fluke in rats. The ability of fluke antigens complexed with sheep antibody to vaccinate rats against infection was also tested. IgM antibodies were involved in precipitate formation on the teguments of fluke 3 weeks after infection but IgG₁ predominated at later stages of infection. The decreased survival of fluke in rats after culture with increasing levels of sheep antibodies suggests that the antibodies exert some deleterious effect on the fluke in vitro. The fluke antigen-sheep antibody complex failed to immunize rats against infection. Since sheep antibodies to F. hepatica can impair the ability of fluke to resist further attack in rats but not sheep, it is suggested that some effector mechanism other than antibody is defective in the latter.     

The stress of Lymnaea truncatula just before miracidial exposure with Fasciola hepatica increased the prevalence of infection.

Single-miracidium infections of Lymnaea truncatula with Fasciola hepatica were carried out under laboratory conditions to determine whether the stress of snails just before miracidial exposure had any influence on the prevalence of Fasciola infection, redial burden, and cercarial shedding. Three methods, i.e., the fasting of L. truncatula for 3 days in water filtered through a Millipore membrane, the effect of 6-8 degrees C water for 15 min, or the immersion of L. truncatula in a detergent solution at low concentration for 15 min, were used to stress snails. Enhanced susceptibility of snails to F. hepatica infection was noted in stressed groups (93-96% vs 48-50% in controls). The number of free rediae did not show any variation in controls as well as in stressed groups, except for fasted snails in which free rediae were significantly fewer. No differences in cercarial production between controls and the cold group were noted. Fasting, cold shock, or detergent exposure prior to exposure to F. hepatica miracidia might have weakened the snails so that they were not as efficient in avoiding miracidial penetration, thus leading to higher infection rates.     

Effect of concurrent infection with Muellerius capillaris on the development of redial generations of Fasciola hepatica in Lymnaea truncatula.

The rediae of Fasciola hepatica were counted according to generation in adult and juvenile Lymnaea truncatula following single infection with Fasciola hepatica, double infection with F. hepatica and then Muellerius capillaris, or double infection with M. capillaris and F. hepatica. The rediae found in double infections were essentially first generation and an early cohort from the second generation. The following differences were observed in adult snails which underwent double infection when compared to corresponding single infections: i) dependent rediae were almost completely absent; ii) degenerating independent rediae were found in identical or decreased numbers; iii) living independent rediae of the first generation were decreased in number, while those of the second generation had variable decreased numbers. The results were similar in juvenile snails with double infections, except that the numbers of degenerating independent rediae were higher than those found in corresponding single infections and the numbers of rediae of the second generation were increased. The order of exposure in double infections had no influence on the number and maturation of fasciolid rediae.     

Effect of the exposure to Fasciola hepatica (Trematoda: Digenea) on life history traits of Lymnaea cousini and Lymnaea columella (Gastropoda: Lymnaeidae)

The snails Lymnaea columella and Lymnaea cousini have both been reported as intermediate hosts of Fasciola hepatica in Colombia. The effect of the exposure to the parasite on survival, fecundity and size of these snails was evaluated by means of experimental infections and the life history traits of control and exposed groups were compared. Infection rates were 82.2 and 34% for L. columella and L. cousini, respectively. A reduction in fitness was observed in both species when exposed to the parasite: fecundity alone was reduced in L. columella whereas in L. cousini there was also a decline in survival rate. Unlike other studies, increased size was not observed in either species. On the contrary, a reduction in growth rate was observed in L. columella.     

Search of natural occurrence of xiphidiocercariae (trematoda) in fresh water snails of nine countries from São Paulo State,Brazil

Xiphidiocercariae, aquatic larval stages of some trematodes are considered a potential instrument for biological control of mosquitoes. In this study we evaluated its natural occurrence in Campinas region and two places in Vale do Ribeira (Registro and Miracatu), S?o Paulo State. Snails were obtained from fresh water collections from September 1996 to February 1999. The species collected were Lymnaea columella, Physa marmorata, Biomphalaria tenagophila, Biomphalaria sp., Drepanotrema cimex, D. lucidum and Drepanotrema sp. Fasciola hepatica, xiphidiocercariae (Haematoloechidae) and echinostomatid cercariae were detected in the lymnaeids snails from Miracatu, SP. In the same locality were found planorbids snails parasitized by furcocercariae, echinostomatid cercariae and xiphidiocercariae. The xiphidiocercariae found in the planorbids were different from those obtained from lymnaeids. One Biomphalaria sp. infected with furcocercariae was found in Louveira, SP. In the ROSA place (Campinas, SP) an individual of Biomphalaria sp. and one of L. columella were found infected by the furcocercariae and echinostomatid cercariae, respectively. In the place UNI-I, in Campinas, one L. columella was infected by furcocercariae. Double infection in snails from Miracatu was also observed.     

First report of Lymnaea cousini Jousseaume, 1887 naturally infected with Fasciola hepatica (Linnaeus, 1758) (Trematoda: Digenea) in Machachi, Ecuador

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 presence of predators modifies the larval development of Fasciola hepatica in surviving Lymnaea truncatula

Experimental infections of Lymnaea truncatula with Fasciola hepatica were performed to study the consequences of the presence of predators (sciomyzid larvae or zonitid snails) on the characteristics of larval F. hepatica development in surviving snails. Controls consisted of infected snails that were not subjected to predators. Compared to controls, the survival rate at day 30 post-exposure, the duration of cercarial shedding, and the number of cercariae shed by surviving snails were significantly lower when predators were present in snail breeding boxes, whatever the type of predator used. In contrast, the prevalences of Fasciola infections in snails, and the length of time between exposure and the onset of cercarial shedding showed no significant variation. The progressive development of a stress reaction in surviving snails against predators during the first 30 days of experimental exposure to F. hepatica would influence snail survival during the cercarial shedding period and, consequently, the number of cercariae shed by the snails.     

Development of Fasciola hepatica in Lymnaea columella infected with miracidia derived from cattle and marmoset infections

The development of Fasciola hepatica from two species of definitive hosts, i.e. cattle (Bos taurus) and a marmoset (Callithrix penicillata) in the snail Lymnaea columella was determined based on the production of rediae and cercariae and snail survival rate. More rediae and cercariae at 60-74 days post-infection were produced by snails infected by cattle-derived miracidia (cattle group) than by those infected by marmoset-derived miracidia (marmoset group). Among the L. columella parasitized by the marmoset group, the survival rate and the percentage of positive snails were higher than among those parasitized by the cattle group. Eggs of F. hepatica released in cattle faeces were significantly bigger than those released in marmoset faeces. Miracidia originating from parasites that completed their development in cattle were more efficient in infecting the intermediate host. These results suggest that vertebrate-host origin influences the eggs produced by the parasite and the infection rates in the snail host L. columella.     

Extremely low frequency magnetic field and the hatching rate of Fasciola hepatica eggs, the fecundity and survival of liver fluke-infected snail, Lymnaea truncatula

Eggs of Fasciola hepatica were exposed for 10 days to extremely low frequency magnetic field (ELFMF) at the frequency of 50 Hz and density of 2 mT (rms). The results show an accelerated hatching of F. hepatica eggs in relation to control (non-exposed) group. The host snails, Lymnaea truncatula, were divided into three groups; those of groups I and II were infected with the miracidia of F. hepatica hatched from control egg culture, whereas those of group III were infected with miracidia hatched from eggs affected by ELFMF. Thereafter, snails of groups II and III were exposed to ELFMF for 53 days, whereas those of group I were not exposed. At day 14 post infection, a significant decrease was observed in the number of cocoons laid by snails of group III, compared with control. Also, significant mortality in group III snails was observed 42 days post infection. The increased mortality and a lower number of cocoons laid by group III snails have probably resulted from enhanced stimulation of metacercarial parthenogenetic reproduction in consequence of infecting the molluscs with miracidia reared under ELFMF.     

Glutathione S-transferase. Novel vaccine against Fasciola hepatica infection in sheep

The potential of GST as a vaccine candidate against liver fluke infection in ruminants was studied by vaccinating sheep (n = 9) with GST purified from adult worms of Fasciola hepatica and challenging with 500 F. hepatica metacercariae. The immunization induced a high antibody response to GST in contrast to the poor or undetectable response to this Ag observed in naturally infected sheep. Throughout the trial, the progress of the fluke infection was monitored by measuring RBC hemoglobin levels, the extent of liver damage and the fecal egg output in the sheep. This analysis indicated that a subpopulation (n = 4) of the GST vaccinated animals exhibited no anemia, reduced liver damage and a lower mean fecal egg count relative to the infected control group suggesting a lower fluke burden in these animals. Worm burdens in the livers of the GST vaccine group (107 +/- 22) were 57% lower than in the infected control group (250 +/- 25). The subpopulation of the GST vaccine group demonstrated a 78% reduction in mean worm burdens relative to the control group. These results show that GST of adult F. hepatica is a novel Ag that can significantly protect sheep against liver fluke infection. The results suggest that the immune response to GST is directed to the juvenile worm reducing the number of worms that can establish in the liver of the vaccinated animals.     

An experimental study of the effects of Nosema eurytremae (Microsporida: Nosematidae) on the liver fluke Fasciola hepatica

A microsporidian parasite, Nosema eurytremae, was fed to Lymnaea trunculata infected with Fasciola hepatica. Microsporidian infection of the snail was always light, though spores were present in all tissues but the rediae became heavily infected in the parenchyma. The proportion of infected rediae increased with time and was especially high in ageing infections. Although rediae were only lightly infected when examined before cercarial release began, the infections became progressively heavier, so that towards the end of the life of the snails a high proportion of rediae were totally destroyed and contained no cercariae. Cercarial output was significantly depressed over a long period of active shedding by snails which had been hyperinfected with 1 x 106 spores and some of the liberated cercariae were themselves infected.     

Comparison of adult liver flukes from highland and lowland populations of Bolivian and Spanish sheep

A morphological study of adult liver flukes and eggs from sheep in a human fascioliasis endemic zone in the Northern Bolivian Altiplano showed that they belong to the species Fasciola hepatica. An exhaustive morphometric comparison with a F. hepatica population from Spanish sheep was made using image analysis and an allometric model: (y2m-y2)/y2 = c(y1m-y1)/y1[b, where y1 = body surface or body length, y2 = one of the measurements analyzed, y1m, y2m = maximum values towards which y1 and y2 respectively tend, and c, b = constants. Only slight allometric differences in worms were observed despite the geographic distance between both Spanish and Bolivian sheep populations and the very high altitude of the Bolivian Altiplano.