Mechanisms of survival of nematode parasites with emphasis on hypobiosis

Mechanisms involved in parasitic nematode survival must be considered with reference to their host and environmental interrelationships since these interrelationhips ultimately influence any parasite adaptations aimed at survival.The most important of the potential environmental constraints are climatic, particularly temperature and humidity, and these can drastically influence larval development and survival.One of the major host factors influencing successful parasite survival is the availability of suitable (susceptible) new hosts to the infective stages of the parasite at the appropriate time for transmission to be achieved. Other host factors that influence parasite survival are those that affect the entrance, establishment and reproduction of the parasite within its new host; mainly problems of acclimatization to a parasitic way of life as well as the countering or adaptation to a variety of host resistance factors, both molecular and cellular, by the parasite.Finally in order for life cycles to be completed, the parasite must evolve means whereby its larval forms can leave the host so that eventual transmission to a new host can be accomplished.In this paper a number of adaptations which enable the parasite to overcome these constraints are discussed. These include such things as larval resistance to environmental effects, the utilization of intermediate hosts or vectors for transmission, seasonally-increased fecundity rates, anti-host immunity stratagems and hypobiosis.This latter phenomenon, hypobiosis or prolonged but temporarily arrested larval development, represents one of the most useful of life cycle adaptations to ensure parasite adaption enables the parasite to synchronize its life cycle to changing environmental or survival and appears to be widespread among parasitic nematodes. Among its benefits, this host conditions. It can thus be of major importance in ensuring survival of the parasite during periods of environmental adversity when conditions for transmission are poor and survival of free-living forms may be minimal. It also enables the parasite to have available large numbers of infective forms at points in the host reproductive cycle which coincide with the production of the susceptible neonates thereby greatly facilitating transmission. Additionally, with certain species of nematodes, occurring as it does at times of the year when the numbers of infective stages may be high while host resources may be limited, oscillations in parasite biomass can be avoided. It thus serves as a highly adaptive mechanism for regulating populations of adult worms, lessening stress on the host and favoring parasite survival as a result.     

Emergence of fasciolosis in cattle in East Anglia

Liver fluke (Fasciola hepatica) infection caused weight loss, diarrhoea, decreased milk yield and occasionally death in cattle in East Anglia during the winters of 2001 to 2003. The condition had previously been limited mainly to stock imported into this part of Britain from endemically infected areas. In composite faecal samples collected by 16 farm animal veterinary practices in Norfolk, Suffolk and Essex, fluke eggs were found in 15 (28.8 per cent) of 52 previously unaffected suckler herds and 10 (16.7 per cent) of 60 dairy herds. Antibodies to F hepatica were detected by elisa in 32 (53.3 per cent) of the bulk milk samples from these 60 dairy herds, including the 10 in which fluke eggs were found. The emergence of fasciolosis in East Anglia was attributed to recent higher summer rainfall, which favoured the intermediate snail host Lymnaea truncatula and the free-living stages of F hepatica, the increased influx of sheep from endemic fluke areas for seasonal grazing, and the wetter grazing conditions associated with the Environmentally Sensitive Area scheme.     

Fasciola hepatica: an assessment on the vectorial capacity of Radix labiata and R. balthica commonly found in Belgium

A previous study conducted in Belgium revealed that genetic material of Fasciola sp. was present in snail species belonging to the genus Radix. Here, these snails were collected and identified by DNA-based techniques as Radix labiata and Radix balthica. These two species and Galba truncatula (the major intermediate host in Europe) were experimentally infected with Fasciola hepatica. The resulting metacercariae were fed to rats and the infection was monitored using several techniques. Microscopy revealed the presence of larval stages in 78.3, 45, and 6.25% of G. truncatula, R. labiata, and R. balthica snails, respectively. These results were confirmed by a PCR that amplifies a Fasciola sp. specific sequence. Furthermore, this PCR was found to be more sensitive than microscopic examination. R. labiata shed fewer metacercariae than G. truncatula but these were as infective to rats as those shed by G. truncatula. This study demonstrates that R. labiata may act as an incidental intermediate host for F. hepatica in Belgium.     

Infection of Lymnaea truncatula and Lymnaea glabra by Fasciola hepatica and Paramphistomum daubneyi in farms of central France

Natural infection of Lymnaea glabra and Lymnaea truncatula by Fasciola hepatica and/or Paramphistomum daubneyi was studied at two periods of risk (June-July and September-October) in 11 French farms known for their high prevalences of F. hepatica infection in ruminants. A total of 1,778 L. truncatula and 2,396 L. glabra measuring 6 mm or more in height were collected to determine the prevalence of natural infection with F. hepatica and P. daubneyi. The role of four factors, i.e. season, snail co-infection (F. hepatica-P. daubneyi), ruminant host and the existence of single or mixed intermediate host communities, was investigated. There were no differences in prevalences between the two risk periods. Co-infections in L. glabra were more frequent than expected. The prevalences of infection with F. hepatica or P. daubneyi in L. glabra were higher in farms rearing sheep than in farms rearing cattle. A similar finding was also noted for L. truncatula infected with P. daubneyi only. L. glabra was a much better intermediate host for F. hepatica and P. daubneyi when it was the only available snail, possibly indicating an adaptation of parasites to their less usual host in local conditions. The two trematodes preferably developed in L. truncatula rather than in L. glabra when both host species lived in the same places.     

Recovery of Fascioloides magna (Digenea) population in spite of treatment programme? Screening of Galba truncatula (Gastropoda, Lymnaeidae) from Lower Austria

During the past decade, Fascioloides magna, the large American liver fluke, has spread within free-living deer in wetlands of the Danube in Lower Austria. The aim of this study was to determine the current infection rates with F. magna and other digenean parasites in the intermediate host snail Galba truncatula from risk areas in Lower Austria. A total of 3444 G. truncatula were collected and examined microscopically for the presence of digenean trematodes. A set of randomly selected snails and isolated trematode stages were also investigated molecular biologically by PCR and sequencing. Digenean parasites were detected with a prevalence of 2.41% (1.83% Paramphistomoidea; 0.46% Echinostomatoidea; 0.09% Strigeida; 0.06% Plagiorchiida). F. magna was found with an overall prevalence of 0.23%, which may indicate a recovery of the parasite population in spite of an ongoing triclabendazole treatment programme. Moreover, high risk areas and a seasonality of infections were observed.     

Soil type and the distribution of Lymnaea truncatula

Habitat distribution of the snail Lymnea truncatula, the intermediate host of the liver fluke Fasciola hepatica, was related to a soil map in a part of Dyfed in South Wales. There was a close association with wet soils in general but, within these areas, habitats were concentrated in very wet localities of restricted size, such as ditches and seepages. Vegetation characteristic of these habitats is described. Soil maps could be a useful tool for the veterinary officer, agricultural adviser and research worker in identifying fluke-prone land.     

Prevalence of Fasciola hepatica in the intermediate host Lymnaea truncatula detected by real time TaqMan PCR in populations from 70 Swiss farms with cattle husbandry

Bovine fasciolosis is an economically important parasitic disease. Quantitative real time PCR was utilized to determine the prevalence of Fasciola hepatica in the snail intermediate host Lymnaea truncatula from 70 selected, infected Swiss cattle farms, and to gain information on the infection risk to the definitive host. Snails from 130 habitats (36 streams, 21 wells, 24 drainage ditches, 33 spring swamps, 14 reeds, 1 drainage shaft and 1 pond) originating from 71 dairy cow pastures, 39 pastures for young stock, 14 hay fields and 6 dry cow pastures were collected. Of these, 51 populations were found to be infected with F. hepatica. A total of 4733 snails were examined of which 331 were infected (7.0%). The numbers of snails collected from different sites ranged from 1 to 159 snails. Clustering of infection in snails was found on the farm of origin with a mixed logistic model with random effects. The risk of infection of L. truncatula with F. hepatica was significantly higher in populations originating from spring swamps, wells and reeds compared to populations from streams. In addition the risk of snail infection was significantly lower in populations collected in young stock and dry cow pastures compared to dairy cow pastures. The greater the population size collected from a habitat also increased the risk of an individual snail being infected.     

Potential for the spread of Fasciola hepatica in cattle in Oklahoma

In 29 central and western Oklahoma counties, 42 ranches were investigated for indigenous Fasciola hepatica infections and their suitability for the transmission of liver fluke. A 10-year retrospective study of Oklahoma cattle based on samples submitted to the Oklahoma Animal Disease Diagnostic Laboratory also was done. Indigenous fascioliasis was found in cattle in 12 counties (9 central and western counties identified in the combined field and retrospective studies and 3 additional eastern counties identified on the basis of the retrospective study). Factors essential for survival of free-living stages of F hepatica and for snail species necessary for propagation of the fluke existed in much of Oklahoma. Snails capable of serving as intermediate hosts for transmission of the fluke reportedly have been found in 41 of the 77 Oklahoma counties. Lymnaeid snails were detected in 4 counties in the field study.     

Ecological basis of parasite control: Trematodes with special reference to fascioliasis

In this paper the main ecological components of selected animal trematode infection cycles are considered with regard to the possibilities of control. The geographical distributions of Fasciola hepatica, F. gigantica, Paramphistomum microbothrium, Schistosoma bovis and S. mattheei are compared in relation to the distribution patterns of their snail intermediate hosts. In order to constitute an effective lifecycle host, intermediate host and the eggs and freeliving stages of the parasite overlap in the effective biotope. Comparing the biotopes of the different trematodiases it becomes evident that they have much in common particularly as the snail intermediate hosts occupy identical niches in a food-chain in which the pollution by the host species results in abundant food for the snail. Moreover, the physical activaties of herbivores, e.g. trampling down the turf, create favourable habitats for the intermediate host.The dynamic nature of the freshwater snail habitats characteristic for trematodiasis, regularly results in a complete disappearance of the effective biotopes in the field. With regard to the parasite these effects are overcome by a long life-expectancy of egg producing parasites in the host population. Small scale changes in availability of appropriate hosts for successful cercarial establishment are compensated by the extended period of cercarial shedding from the snail population.These features in combination with the survival of eggs and the grazing and drinking behaviour characteristics of the hosts tend to stabilize the potential oscillations in transmission rate. Because of the complicated nature of interdependency within the host-trematode-intermediate host complex, the ecological base with regard to practical measures is fundamentally wide. However, the pressure upon the situation by social and economic conditions limits operative possibilities considerably.     

Epidemiology of gastrointestinal nematodes of sheep and goats in Haryana, India

Epidemiological studies on some gastrointestinal nematodes of sheep and goats in Karnal, Ambala and Rohtak districts of Haryana (India) revealed that Haemonchus contortus and Trichostrongylus spp. were responsible for parasitic gastroenteritis in these hosts. The adult parasites persisted in the host throughout the year and there was no indication of hypobiosis. The maintenance of the parasite population depended on a continuous cycle of infection between the host and the pasture. The agroclimatic conditions of the districts revealed that, in general, favourable weather conditions for the development and survival of the free-living stages of the parasites existed all year round.     

The distribution of Fasciola hepatica in Queensland, Australia, and the potential impact of introduced snail intermediate hosts

A survey was conducted to establish the distribution of the liver fluke, Fasciola hepatica, in the state of Queensland, Australia, and to evaluate the impact of the introduced snail intermediate hosts, Pseudosuccinia columella and Austropeplea viridis. Serum samples from a total of 5103 homebred cattle in 142 beef herds distributed throughout the state and 523 pooled milk samples from dairy herds from the state's major dairying regions were tested for antibodies to F. hepatica by ELISA. Snails were collected on infected properties around the limits of the F. hepatica distribution. F. hepatica infection was detected in 44 dairy herds and two beef herds. The distribution of infected herds indicates that F. hepatica is established only in southeast Queensland. The distribution there was patchy but the parasite was more widespread than suggested by an earlier survey. The predominant intermediate host species found along the northern limit of the distribution was P. columella. We conclude that the introduction of P. columella and A. viridis has not yet had a major impact on the distribution of F. hepatica in Queensland. However, the presence of P. columella, which is much more adaptable to tropical habitats than the native intermediate host, Austropeplea tomentosa, at the northern limit of the F. hepatica distribution suggests that there is potential for the parasite to expand its range.     

Epizootiology of fascioliasis in Montana.

During 1989-1990, the United States Department of Agriculture (USDA) meat inspection records were used to determine the distribution and incidence of liver flukes (Fasciola hepatica and Fascioloides magna) in Montana cattle. Of the cows and bulls slaughtered in USDA-inspected packing plants during a 12 month time period, 17.24% had livers that were condemned because of liver flukes. This was a 12% increase over USDA liver condemnations reported for 1973. Infected animals have been reported from 26 counties in Montana, mostly located in the south-central and western half of the state. Forty-nine percent of the 2.4 million cattle in Montana are raised in these counties. Lymnaeid snail species that may serve as intermediate hosts for Fasciola hepatica were found in most of the counties where liver flukes were reported. The principal vectors believed to be responsible for the transmission of Fasciola hepatica in Montana are species of the genus Fossaria. Stagnicola montanensis and Lymnaea stagnalis, which may serve as intermediate hosts for this parasite have also been collected. A known intermediate host for Fascioloides magna, Stagnicola caperata, was also found in several locations.     

Temporal studies on Fasciola hepatica in Galba truncatula in the west of Ireland

The population dynamics and prevalence of Fasciola hepatica in Galba truncatula were investigated on the Teagasc hill sheep farm in Country Mayo, Ireland from August 2006 until March 2008. Galba truncatula (n=974) were collected at fortnightly intervals in 4 snail habitats on the farm, their relative density and mean monthly rainfall and temperature were recorded. Snail abundance was associated with rainfall and temperature. G. truncatula abundance was generally low during winter 2006/2007 (December 2006-February 2007), mid spring and summer 2007 (April-July) and late autumn and winter 2007/2008 (October 2007-February 2008). Overall, two peaks in G. truncatula abundance were evident during the study period, an early spring (March) and late summer/autumn (late August-October) peak. Seasonal differences were observed in the prevalence of F. hepatica in G. truncatula, the greatest prevalence was evident in 2007 during the summer (25%) and autumn (16%). Two seasonal transmission peaks, as evident by the presence of mature infections in snails, one in summer/autumn and the other in late winter/early spring. Not unsurprising, cercariae (28%) and rediae (29%) were the predominant developmental stages observed in mature and pre-adult snails respectively. Worthy of note in this study, cercariae were also evident in pre-adult snails. The present study highlights the influence of milder temperatures and wetter conditions on both snail and fluke development. If the trend in mild and wet weather conditions continues, there is the potential for an increase in the number of G. truncatula habitats and ultimately the prevalence of fasciolosis in grazing livestock in Ireland.     

Cathepsin L proteinases as vaccines against infection with Fasciola hepatica (liver fluke) in ruminants

Fasciolosis, caused by trematodes of the genus Fasciola, is one of the most important diseases of farmed ruminants in temperate and tropical zones. The appearance of Fasciola hepatica populations that are resistant to common flukicidal drugs means that new methods of treatment will soon be required. The future prospect for the development of anti-liver fluke vaccines is optimistic and given their consumer acceptability and environmental friendliness, offer the best way forward. Cathepsin L proteases (Fhe CL 1 and Fhe CL 2), secreted by liver flukes at all stages of their development in the mammalian host, are believed to play important roles in facilitating parasite migration (tissue degradation), feeding and immuno-evasion. The authors consider them prime targets for which new vaccines can be developed. Vaccine studies in cattle and sheep have shown that protection levels of up to 72 and 79 per cent, respectively, can be obtained with immunisations of cathepsin Ls in Freunds' adjuvant. The vaccine also exhibited high anti-embryonation/anti-fecundity effects on parasites that survived in vaccinated animals and thus could have a major impact on the transmission of disease to the intermediate host. While natural infections in sheep and cattle appear to elicit non-protective Th2 immune responses, the authors' studies indicate that the protection induced by vaccination involves elements of a Th1 response.     

PCR diagnosis of Fasciola hepatica in field-collected Lymnaea columella and Lymnaea viatrix snails

Fasciolosis, caused by the trematode Fasciola hepatica, is a zoonosis of economic importance in livestock that is emerging as a chronic disease in humans. The intermediate hosts are lymnaeid snails, in which diagnosis of infection is traditionally based on cercarial shedding, tissue sectioning and crushing. We developed a PCR assay for the sensitive and specific detection of F. hepatica in field-collected Lymnaea sp. snails. A primer pair was designed to amplify a 405 bp fragment of the cytochrome c oxidase subunit 1 gene of F. hepatica. The PCR assay showed a limit of detection of 10 pg of genomic F. hepatica DNA. No cross-reactions were observed with samples from other related trematode species or from the snail hosts Lymnaea columella and Lymnaea viatrix. DNA sequencing of the amplicon showed 100% homology with F. hepatica, and 75-89% homology with other trematodes on regions that did not include the entire set of primers. Two samples from Argentina were analysed. For snails in sample 1 (n = 240), identified as L. columella, the infection rate was 17.5 and 51.3% by direct examination and PCR, respectively. For snails in sample 2 (n = 34), identified as L. viatrix, the infection rate was 2.9 and 61.8% by direct examination and PCR, respectively. Differences in infection rates between these diagnosis methods were significant for both samples. Our PCR technique showed to be effective for detecting specific F. hepatica infections of low intensity in the intermediate host, and hence it could be used to study the epidemiological situation in a given area, as well as to assess host suitability for the parasite.     

Fasciola hepatica products induce apoptosis of peritoneal macrophages

Immunomodulatory properties have been described for Fasciola hepatica excretory-secretory products (FhESP), with their interaction with the innate immune cells being crucial during the early stages of infection. Previously, we demonstrated that FhESP induce eosinophil apoptosis. In this work, the ability of FhESP to induce apoptosis of peritoneal macrophages was evaluated. These parasite products were observed to induce apoptosis in peritoneal macrophages stimulated in vitro with FhESP, as well as in cells recovered from infected mice. The ability of FhESP to modify the viability of macrophages by apoptosis induction may constitute a crucial event for extending its survival in the host.     

The influence of different food sources on cercarial production in Lymnaea truncatula experimentally infected with Digenea

Experimental infections of Lymnaea truncatula with a digenean species (Fasciola gigantica, F. hepatica, or Paramphistomum daubneyi) were performed under laboratory conditions to study the effect of four sources of food (microalgae, romaine lettuce, wheat germs, or modified Boray diet) on cercarial production. The mean number of F. hepatica, F. gigantica, or P. daubneyi cercariae was significantly greater when snail food was microalgae or Boray diet. The lowest cost prices for 100 metacercariae of F. hepatica (9.7-10.1 euros) were noted when algae of Boray diet were used as food for snails. In contrast, the highest prices (18.0-18.5 euros) were found in lettuce-reared snails. The choice of a source of food for breeding the intermediate hosts of F. hepatica and collecting metacercariae must take into account the importance of the commercial demand for metacercariae and will depend on the strategy the producer wishes to develop.     

Aspects of the biology of Ostertagia ostertagi in relation to the genesis of ostertagiasis

Adaptations such as the capacity of free-living stages to survive environmental stress, inhibition and density-dependent effects on population size and fecundity show O. ostertagi to be a consummate parasite of cattle in temperate environments. Knowledge of these adaptations, within an animal management context, provides the key to understanding the occurrence of disease and a basis for control of parasite numbers. Substantial infections do arise from low egg contamination rates of pastures. Discontinuities in rates of infection are caused by poorly predicted seasonal events such as the effects on eggs of oxygen-deficient environments, fluctuating temperatures and the absence of sufficient moisture for migration of infective larvae from faeces to herbage. Time delays of several months between pasture contamination and availability of infection are therefore common. Ingestion of large numbers of larvae, over a short period early in the grazing season, gives rise to Type I disease or subclinical infections which decrease liveweight gains. In some ecotypes, environmentally induced inhibition leads to the accumulation of large populations of early 4th stage parasites within the host. These populations, under poorly defined conditions, can mature synchronously to produce severe diarrhoea, debility and even death in a proportion of mature cattle--the Type II disease.     

Dynamics of Fasciola hepatica infection in two species of snails in a rural locality of Mexico

The aim of the present study was to identify the species that are natural intermediate hosts of Fasciola hepatica in a Research Centre in Chapa de Mota, State of Mexico, Mexico, where temperate and humid conditions prevail. Also, the magnitude and dynamics of the natural infection in four biotopes were analysed. The study was performed between October 2000 and November 2001. The molluscs were collected, identified and studied for trematode infection. Soil humidity and temperature were determined monthly. A total of 4042 snails were collected 3372 (83%) were Fossaria humilis and 670 (17%) F. bulimoides. F. hepatica was present in 2537 (75.2%) and 515 (76.9%) of the two snail species, respectively. Temperature, soil humidity and microenvironment affected the dynamics of the host population and the parasite infestation, with a significant increase from August to November 2001. The highest prevalence was observed in the warmer and more humid biotopes.