The potential of Capillaria hepatica to control mouse plagues

The potential of helminths as bio-control agents of mammalian pests has been largely ignored. However, the nematode Capillaria hepatica is currently being examined for its potential to control population outbreaks of house mice in Australia. Grant Singleton and Hamish McCallum discuss laboratory and ecological studies of the parasite and host, and describe the results of two models that explore the interaction between C. hepatica and mouse populations.     

Changes in parasite transmission stage excretion after pheasant release

The production of parasite transmission stages was investigated in the faeces of 77 farm-bred ring-necked pheasants (Phasianus colchicus). Coccidian oocysts (Eimeria sp.), and nematode eggs (Heterakis sp., and Capillaria-like eggs) were recovered before and after release but all birds were treated prior to release. Treatment with fenbendazole significantly reduced the abundance of transmission-stage excretion for all parasites, and reduced the prevalence in the case of Eimeria sp. and Heterakis sp. Nonetheless, a significant increase in the excretion abundance for all parasites and in the prevalence of Eimeria sp. and Heterakis sp. was found after release. Eggs of Ascaridia sp. were found only after releasing, suggesting infection ocurred in the wild. A negative relationship was found between the pheasant body condition and Heterakis excretion abundance and a higher abundance of Capillaria sp. eggs in female birds. No significant relationship was found between parasite excretion abundance and pheasant survival. Despite this, results suggest that an increase in the excretion of parasite transmission stages follows the release of captive pheasants into the wild. This can in part explain restocking failures, but also means that autochtonous free-living birds may become exposed to new and potentially harmful pathogens. To avoid these risks it is proposed that improved prophylactic measures should be taken.     

Parasites of European hedgehogs (<Emphasis Type="BoldItalic">Erinaceus europaeus</Emphasis>) in Britain: epidemiological study and coprological test evaluation

Seventy-four European hedgehogs (Erinaceus europaeus) that had died in wildlife rehabilitation centres were dissected and their parasite burdens documented. Overall parasite prevalence was 91%, and a total of six helminth species were isolated: five nematodes (Crenosoma striatum, Eucoleus aerophilus, Capillaria erinacei, Capillaria ovoreticulata and Capillaria spp.), one trematode (Brachylaemus erinacei) and one acanthocephalan (Oliganthorhynchus erinacei). The tick Ixodes hexagonus and flea Archeopsylla erinacei were also collected. The effect of parasite infection on body condition was assessed by correlation of burdens with the residuals of weight–skeletal length regression. Tick presence was positively related to body condition; for other parasites, no significant relationship was found. Faecal egg or larval count was closely correlated with adult parasite burden for C. striatum and Capillaria/Eucoleus spp., but not for other species. Coprological analysis should therefore be useful for in vivo studies of nematode parasite infection in hedgehogs. The epidemiology of parasites in hedgehogs and their possible role in recent population declines are discussed.     

RNA interference and plant parasitic nematodes

RNA interference (RNAi) has recently been demonstrated in plant parasitic nematodes. It is a potentially powerful investigative tool for the genome-wide identification of gene function that should help improve our understanding of plant parasitic nematodes. RNAi should help identify gene and, hence, protein targets for nematode control strategies. Prospects for novel resistance depend on the plant generating an effective form of double-stranded RNA in the absence of an endogenous target gene without detriment to itself. These RNA molecules must then become available to the nematode and be capable of ingestion via its feeding tube. If these requirements can be met, crop resistance could be achieved by a plant delivering a dsRNA that targets a nematode gene and induces a lethal or highly damaging RNAi effect on the parasite.     

Some differences in the parasitic fauna of three samples of plaice (Pleuronectesplatessa L.) from the southern North Sea

Plaice ( Pleuronectesplatessa L.) from three spawning areas in the southern North Sea were examined for six species of metazoan parasite. Differences of both incidence and level of infestation with particular parasite species were found in fish from the three areas. Neither the incidence nor the level of infestation was associated with the length of fish in the samples except in the case of the larger male fish from Flamborough, which tended to contain more larval nematodes (Anisakidae). All fish contained the intestinal nematode Cucullanus heterochrous , although fewer were found in fish from Flamborough. Southern Bight fish were more likely to contain Capillaria wickinsi than were fish from Flamborough or the German Bight. Flamborough fish were more likely to contain Anisakidae larvae than were fish from the other two areas. Fish from the three areas were similarly infested with Zoogonoides viviparus. Male fish from Flamborough were less likely to be infested with Lepeophtheirus pectoralis than male fish from the German Bight.     

Exploring the Gastrointestinal “Nemabiome”: Deep Amplicon Sequencing to Quantify the Species Composition of Parasitic Nematode Communities

Parasitic helminth infections have a considerable impact on global human health as well as animal welfare and production. Although co-infection with multiple parasite species within a host is common, there is a dearth of tools with which to study the composition of these complex parasite communities. Helminth species vary in their pathogenicity, epidemiology and drug sensitivity and the interactions that occur between co-infecting species and their hosts are poorly understood. We describe the first application of deep amplicon sequencing to study parasitic nematode communities as well as introduce the concept of the gastro-intestinal “nemabiome”. The approach is analogous to 16S rDNA deep sequencing used to explore microbial communities, but utilizes the nematode ITS-2 rDNA locus instead. Gastro-intestinal parasites of cattle were used to develop the concept, as this host has many well-defined gastro-intestinal nematode species that commonly occur as complex co-infections. Further, the availability of pure mono-parasite populations from experimentally infected cattle allowed us to prepare mock parasite communities to determine, and correct for, species representation biases in the sequence data. We demonstrate that, once these biases have been corrected, accurate relative quantitation of gastro-intestinal parasitic nematode communities in cattle fecal samples can be achieved. We have validated the accuracy of the method applied to field-samples by comparing the results of detailed morphological examination of L3 larvae populations with those of the sequencing assay. The results illustrate the insights that can be gained into the species composition of parasite communities, using grazing cattle in the mid-west USA as an example. However, both the technical approach and the concept of the ‘nemabiome’ have a wide range of potential applications in human and veterinary medicine. These include investigations of host-parasite and parasite-parasite interactions during co-infection, parasite epidemiology, parasite ecology and the response of parasite populations to both drug treatments and control programs.     

In silico analyses of neuropeptide-like protein (NLP) profiles in parasitic nematodes

Nematode parasite infections cause disease in humans and animals and threaten global food security by reducing productivity in livestock and crop farming. The escalation of anthelmintic resistance in economically important nematode parasites underscores the need for the identification of novel drug targets in these worms. Nematode neuropeptide signalling is an attractive system for chemotherapeutic exploitation, with neuropeptide G-protein coupled receptors (NP-GPCRs) representing the lead targets. In order to successfully validate NP-GPCRs for parasite control it is necessary to characterise their function and importance to nematode biology. This can be aided through identification of receptor activating ligand(s) via deorphanisation. Such efforts require the identification of all neuropeptide ligands within parasites. Here we mined the genomes of nine therapeutically relevant pathogenic nematodes to characterise the neuropeptide-like protein complements and demonstrate that: (i) parasitic nematodes possess a reduced complement of neuropeptide-like protein-encoding genes relative to Caenorhabditis elegans; (ii) parasite neuropeptide-like protein profiles are broadly conserved between nematode clades; (iii) five Ce-nlps are completely conserved across the nematode species examined; (iv) the extent and position of neuropeptide-like protein-motif conservation is variable; (v) novel RPamide-encoding genes are present in parasitic nematodes; (vi) novel Allatostatin-C-like peptide encoding genes are present in both C. elegans and parasitic nematodes; (vii) novel neuropeptide-like protein families are absent in C. elegans; and (viii) highly conserved nematode neuropeptide-like proteins are bioactive. These data highlight the complexity of nematode neuropeptide-like proteins and reveal the need for nomenclature revision in this diverse neuropeptide family. The identification of neuropeptide-like protein ligands, and characterisation of those with functional relevance, advance our understanding of neuropeptide signalling to support exploitation of the neuropeptidergic system as an anthelmintic target.     

Further studies on Capillaria philippinensis: development of the parasite in the Mongolian gerbil

Capillaria philippinensis larvae from the digestive tract of a Northern Luzon freshwater fish (Hypselotris bipartita) experimentally exposed to embryonated eggs, were given by stomach tube to Mongolian gerbils (Meriones unguiculatus). The larvae developed into adults within 10 to 11 days and female worms produced larvae within 13 to 14 days. These larvae developed into second generation adults by days 22 to 24 and the second generation females produced eggs that were present in the feces of the animal on the average 26 days after infection. Most females were oviparous but a few larviparous females were always present. The gerbils died on an average of 46 days after infection, with the highest numbers of worms recovered between days 36 and 46. All stages of the parasite were generally found at necropsy. Gerbils developed patent infections after receiving 2 or 3 laeval from fish, and 852 to 5,353 worms were recovered at necropsy. These studies show that autoinfection in an integral part of the life cycle of C. philippinensis, both initially and in maintaining the infection. The natural transmission of the parasite was demonstrated when H. bipartita from a lagoon in the endemic area were fed to gerbils and 3 became infected. The parasite can also be maintained in the laboratory by transfer of worms by stomach tube from the small intestines of an infected gerbil to a clean gerbil.     

Molecular characterization of Calodium hepaticum in grey dwarf hamster (Cricetulus migratorius)

Calodium hepaticum (Bancroft, 1893) Moravec, 1982 (Syn. Capillaria hepatica) is a zoonotic nematode that causes hepatic capillariosis, an uncommon zoonotic infection. The nematode is globally distributed and parasitizes the liver of mammals, mainly Muroidea. Cricetulus migratorius Pallas, 1773 (Cricetinae) was rarely reported as a host for C. hepaticum. In Turkey, C. hepaticum was recorded in three rodent species; Rattus rattus, R. norvegicus, and Apodemus flavicollis. In this study, C. migratorius (grey dwarf hamster) has been identified as a new host species for C. hepaticum in Turkey. The parasite was identified by morphological, histological, and molecular methods and the phylogenetic relationships of C. hepaticum collected from different hosts were revealed. This is the first molecular characterization of C. hepaticum from a grey dwarf hamster.     

Heritable transgenesis of Parastrongyloides trichosuri: a nematode parasite of mammals

Germline transformation of a parasitic nematode of mammals has proven to be an elusive goal. We report here the heritable germline transformation of Parastrongyloides trichosuri, a nematode parasite whose natural hosts are Australian possums of the genus Trichosurus. This parasite can undergo multiple free-living life cycles and these replicative cycles can be maintained indefinitely in the laboratory. Transformation was achieved by microinjection of DNA into the ovary syncytium of either free-living or parasitic adult females. By selecting for the transgenic progeny of successive free-living life cycles, it was possible to establish and maintain transgenic lines. All three transgenic lines tested were shown capable of establishing patent infections in possums and to transmit the functional transgene to their progeny. The transgene, driven by the Pt hsp-1 promoter, was constitutively expressed in intestinal cells at all stages of both parasitic and free-living life cycles, although gene silencing appears to occur in some transgenic progeny. This is the first report of heritable transgenesis in a parasitic nematode of a mammal and we discuss a variety of previously inaccessible experimental avenues that will now be possible with this powerful model system.     

Effects of precipitation on parasite burden along a natural climatic gradient in southern Africa – implications for possible shifts in infestation patterns due to global changes

As a consequence of environmental change, it is expected that shifts in temperature and precipitation patterns will influence parasite communities and their hosts with unpredictable impact. Parasites play a vital role in ecosystems but there is only limited quantitative data which describe the effects of environmental parameters under natural conditions. We investigated the influence of rainfall, relative humidity and temperature on the prevalence, abundance and infection intensity of nematodes in southern Africa by studying the gastro‐intestinal helminth community of the striped mouse Rhabdomys pumilio. Along a precipitation gradient from the Cape of South Africa to northern Namibia we trapped 470 mice over a geographical distance of about 1400 km. Faecal egg counts of 439 sampled individuals and dissections of 161 gastro‐intestinal tracts revealed 15 different helminth species. The most abundant nematode species harboured in 62.6% of all infected mice were the oxyurid Syphacia obvelata followed jointly by two species (Heligmonina spira and Neoheligmonella capensis) of the subfamily Nippostrongylinae (43.7%). We found a significant positive correlation between mean annual precipitation (rainfall and relative humidity) and nematode infestation rates of animals and a negative correlation with temperature. In addition, we found associations between precipitation and different qualitative measurements of parasite burden (mean nematode species richness, mean number of nematode worms and infection intensity per individual host). The similarity in nematode species composition decreased with distance between all study sites. Our study indicates for the first time an association between climatic variables and parasite prevalence and abundance along a continuous natural climatic gradient in a small mammal. These results might be incorporated in the development of models which can predict possible threats for the balance of ecosystems and shifts in infestation patterns due to global changes.     

Virulence and competitive ability in an obligately killing parasite

Mixed infections are thought to have a major influence on the evolution of parasite virulence. During a mixed infection, higher within‐host parasite growth is favored under the assumption that it is critical to the competitive success of the parasite. As within‐host parasite growth may also increase damage to the host, a positive correlation is predicted between virulence and competitive success. However, when parasites must kill their hosts in order be transmitted, parasites may spend energy on directly attacking their host, even at the cost of their within‐host growth. In such systems, a negative correlation between virulence and competitive success may arise. We examined virulence and competitive ability in three sympatric species of obligately killing nematode parasites in the genus Steinernema. These nematodes exist in a mutualistic symbiosis with bacteria in the genus Xenorhabdus. Together the nematodes and their bacteria kill the insect host soon after infection, with reproduction of both species occurring mainly after host death. We found significant differences among the three nematode species in the speed of host killing. The nematode species with the lowest and highest levels of virulence were associated with the same species of Xenorhabdus, indicating that nematode traits, rather than the bacterial symbionts, may be responsible for the differences in virulence. In mixed infections, host mortality rate closely matched that associated with the more virulent species, and the more virulent species was found to be exclusively transmitted from the majority of coinfected hosts. Thus, despite the requirement of rapid host death, virulence appears to be positively correlated with competitive success in this system. These findings support a mechanistic link between parasite growth and both anti‐competitor and anti‐host factors.     

The occurrence of black spots in the tongue sole, Cynoglossus browni Chabanaud, due to nematode eggs (Capillaria spinosa) previously described in the shark Carcharhinus mttberti Müller & Henle

The occurrence of black spots in fillets of the tongue sole, Cynoglossus browni , due to the occurrence of nematode eggs is reported. The eggs were found to be identical with Capillaria spinosa egg type previously described from the shark, Carcharhinus milberti . The presence of the capillarid eggs in the tongue sole is discussed as a new observation in relation to gaps in knowledge of the adult nematode and the biology of the shark.     

Severe parasitism in an opossum.

Chronic debilitation and anemia were observed in a free-living opossum (Didelphis marsupialis) heavily parasitized by Physaloptera turgida, Brachylaima virginianum, and Cruzia americana. Chronic interstitial pneumonia associated with Capillaria aerophila and a metastrongyloid nematode also was present.     

Parasite abundance contributes to condition‐dependent dispersal in a wild population of large herbivore

Parasite abundance has been shown to have major consequences for host fitness components such as survival and reproduction. However, although natal dispersal is a key life history trait, whether an individual's decision to disperse or not is influenced by the abundance of parasites it carries remains mostly unknown. Current and opposing hypotheses suggest that infected individuals should either be philopatric to avoid the energetic costs of dispersal (condition dependence) or disperse to escape from heavily parasitised habitats. From intensive monitoring of a roe deer population inhabiting a multi‐use and spatially heterogeneous agricultural landscape, we evaluated the link between an individual's parasite abundance and its propensity to disperse, while accounting for confounding effects of body mass. Dispersal propensity generally decreased with both increasing nematode abundance and with decreasing body mass. Within the dispersing segment of the population, individuals with high nematode abundance left their natal home range later in the season than less parasitised deer. These results clearly show that parasite abundance is an important component of condition‐dependent dispersal in large herbivores. However, unexpectedly, three individuals that were both heavily parasitised and of low body mass dispersed. We suggest that this ‘leave it’ response to high parasite levels in the natal habitat could represent a last ditch attempt to improve reproductive prospects, constituting a form of emergency life history strategy.     

Monkeys in the Middle: Parasite Transmission through the Social Network of a Wild Primate

In wildlife populations, group-living is thought to increase the probability of parasite transmission because contact rates increase at high host densities. Physical contact, such as social grooming, is an important component of group structure, but it can also increase the risk of exposure to infection for individuals because it provides a mechanism for transmission of potentially pathogenic organisms. Living in groups can also create variation in susceptibility to infection among individuals because circulating levels of immunosuppressive hormones like glucocorticoids often depend on an individual’s position within the group’s social structure. Yet, little is known about the relative roles of socially mediated exposure versus susceptibility in parasite transmission among free-living animal groups. To address this issue, we investigate the relationship between host dominance hierarchy and nematode parasite transmission among females in a wild group of Japanese macaques (Macaca fuscata yakui). We use social network analysis to describe each individual female’s position within the grooming network in relation to dominance rank and relative levels of infection. Our results suggest that the number of directly-transmitted parasite species infecting each female, and the relative amount of transmission stages that one of these species sheds in faeces, both increase with dominance rank. Female centrality within the network, which shows positive associations with dominance hierarchy, is also positively associated with infection by certain parasite species, suggesting that the measured rank-bias in transmission may reflect variation in exposure rather than susceptibility. This is supported by the lack of a clear relationship between rank and faecal cortisol, as an indicator of stress, in a subset of these females. Thus, socially mediated exposure appears to be important for direct transmission of nematode parasites, lending support to the idea that a classical fitness trade-off inherent to living in groups can exist.     

Anthelmintic treatment alters the parasite community in a wild mouse host

Individuals are often co-infected with several parasite species, yet the consequences of drug treatment on the dynamics of parasite communities in wild populations have rarely been measured. Here, we experimentally reduced nematode infection in a wild mouse population and measured the effects on other non-target parasites. A single oral dose of the anthelmintic, ivermectin, significantly reduced nematode infection, but resulted in a reciprocal increase in other gastrointestinal parasites, specifically coccidial protozoans and cestodes. These results highlight the possibility that drug therapy may have unintended consequences for non-target parasites and that host–parasite dynamics cannot always be fully understood in the framework of single host–parasite interactions.     

Estimating transmission potential in gastrointestinal nematodes (order: Strongylida)

Microparasite virulence (the potential to cause harm in the host) is thought to be regulated by a direct trade-off with pathogen transmission potential, but it is unclear whether similar trade-offs occur in macroparasites (helminths). In this analysis, the transmission potentials of 5 nematode species (order Strongylida), known to differ in their virulence, were estimated using an index based on egg production and larval survivability. Virulence estimates were based on the minimum number of worms that cause host death. In nematode species where mature adults cause pathology (trichonematidic development), there is a direct relationship between virulence and transmission, suggesting that high virulence is related to parasite fitness in these worms. However, in nematodes where the juvenile stages produce pathology during migration and development (strongylidic development), virulence is not correlated with transmission. These data suggest that trade-offs between transmission and virulence in nematode parasites are not analogous for all species and may depend on the developmental strategy and mechanism of pathogenicity of the parasites.     

Possible commercial formulations of insect-parasitic nematodes

At least two experimental nematode biological insecticide formulations are currently being produced in small quantities around the world, These formulations are efficacious and people are willing to pay money for them. Advantages for the use of these parasites can be: nonpolluting, self-perpetuating, safe, and not harmful to beneficial organisms. The disadvantages are: relatively short shelf-life, acceptability, moisture, price, and dispersal techniques. One of the preparations is a nematode that vectors a bacterium. It can be produced at the rate of 100 × 10⁶ infective nematodes/2 liter container at a cost of 2 cents/10⁶. It can kill over 1000 species of insects. The other product kills over 60 species of mosquitoes and has been sold as mermithid nematode eggs in moist sand. This mosquito parasite can be established in the new site and self-perpetuate to suppress mosquito parasite can be established in the new site and self-perpetuate to suppress mosquito populations. It kills the mosquito before pupating. Increased research activity, using these parasites in the last ten years, has stimulated interest in these organisms for pest insect control.     

An experimental type II mixed cryoglobulinemia with renal glomerulopathy in ICR mice triggered by Capillaria hepatica infection

Type II mixed cryoglobulinemia is characterized by systemic vasculitis with deposition of cryoprecipitatable-immunoglobulins containing rheumatoid factor. Pathogenesis of type II mixed cryoglobulinemia has not yet been completely clarified because of the lack of an experimental animal. Here, we report an animal model of type II mixed cryoglobulinemia that is induced by experimental infection with Capillaria hepatica in ICR mice. Capillaria hepatica is a nematode that causes necrotic hepatitis in several mammals. In this study, mice experimentally infected with C. hepatica eggs developed cryoglobulinemia at 20 and 30 days post injection. Using immunological analysis, cryoglobulinemia in infected mice was classified as type II mixed cryoglobulinemia by detection of monoclonal IgM rheumatoid factor and IgA in the cryoprecipitate of serum. Using immunofluorescence, we observed an increase in the number of double-positive cells for μ heavy and κ light chains of immunoglobulin in the spleens of infected mice. Histopathologically, this model was characterized by glomerulopathy associated with intense deposition of IgM and IgA filling in capillary lumina. Ultrastructural analysis showed that glomerular deposits consisted of stacks of twisted microtubular structures. These serological and histological features resembled those of type II mixed cryoglobulinemia in human. This is the first experimental animal model of type II mixed cryoglobulinemia that will enable detailed studies on the pathogenesis of cryoglobulinemia.