Eye Fluke Diplostomum Pseudospathaceum Research Articles

Trematode Diplostomum pseudospathaceum inducing differential immune gene expression in sexual and gynogenetic gibel carp (Carassius gibelio): parasites facilitating the coexistence of two reproductive forms of the invasive species.

Parasite-mediated selection is considered one of the potential mechanisms contributing to the coexistence of asexual-sexual complexes. Gibel carp (Carassius gibelio), an invasive fish species in Europe, often forms populations composed of gynogenetic and sexual specimens. The experimental infection was induced in gynogenetic and sexual gibel carp using eye-fluke Diplostomum pseudospathaceum (Trematoda), and the transcriptome profile of the spleen as a major immune organ in fish was analyzed to reveal the differentially expressed immunity-associated genes related to D. pseudospathaceum infection differing between gynogenetic and sexual gibel carp. High parasite infection was found in gynogenetic fish when compared to genetically diverse sexuals. Although metacercariae of D. pseudospathaceum are situated in an immune-privileged organ, our results show that eye trematodes may induce a host immune response. We found differential gene expression induced by eye-fluke infection, with various impacts on gynogenetic and sexual hosts, documenting for the majority of DEGs upregulation in sexuals, and downregulation in asexuals. Differences in gene regulation between gynogenetic and sexual gibel carp were evidenced in many immunity-associated genes. GO analyses revealed the importance of genes assigned to the GO terms: immune function, the Notch signaling pathway, MAP kinase tyrosine/threonine/phosphatase activity, and chemokine receptor activity. KEGG analyses revealed the importance of the genes involved in 12 immunity-associated pathways - specifically, FoxO signaling, adipocytokine signaling, TGF-beta signaling, apoptosis, Notch signaling, C-type lectin receptor signaling, efferocytosis, intestinal immune network for IgA production, insulin signaling, virion - human immunodeficiency virus, Toll-like receptor signaling, and phosphatidylinositol signaling system. Our study indicates the limited potential of asexual fish to cope with higher parasite infection (likely a loss of capacity to induce an effective immune response) and highlights the important role of molecular mechanisms associated with immunity for the coexistence of gynogenetic and sexual gibel carp, potentially contributing to its invasiveness.

Enriched rearing environment enhances fitness-related traits of salmonid fishes facing multiple biological interactions

To improve stocking success in threatened populations, captive-reared animals are often familiarized to natural environment. However, whether such actions improve the ability to cope with multiple biological interactions, such as competition, predation, and parasitism that impose contradicting pressures on decision-making, is not well understood. Here, we explored short-term (4 weeks) and long-term (10 months) effects of enriched rearing on fitness-related traits of Atlantic salmon ( Salmo salar). Salmon with different backgrounds of enrichment and parasite infection (eye fluke Diplostomum pseudospathaceum) were released to semi-natural ponds and monitored for activity, growth, and predation susceptibility. Fish from enriched rearing showed lower short-term activity and higher short-term growth, suggesting that they coped better with novel conditions. However, predation susceptibility and longer-term growth and survival were unaffected by rearing treatment. Importantly, parasitism did not remove the positive effect of enrichment on growth, although the infection decreased both short-term and long-term growth and survival. These results suggest that enriched rearing can enhance fitness-related traits, such as growth, of stocked fish, particularly during the critical early days, which can have important implications for stock enhancement activities.

Responses towards eyefluke (Diplostomum pseudospathaceum) in different genetic lineages of rainbow trout.

Marker-assisted selective breeding of fish with higher levels of resistance towards specific pathogens may improve fish health, but the impact of host genotype on susceptibility to multiple pathogen infections is still poorly investigated. This study examined the resistance in rainbow trout Oncorhynchus mykiss towards infection with the eye fluke Diplostomum pseudospathaceum. We used genetically selected rainbow trout, carrying SNPs associated with resistance towards the parasitic ciliate Ichthyophthirius multifiliis, and exposed the fish to eye fluke cercariae. We showed that fish partly resistant to I. multifiliis were more susceptible to eye fluke invasion. The expression of immune relevant genes (encoding innate and adaptive factors) was also affected as these genotypes responded less strongly to a secondary fluke infection. The complexity of genome architecture in disease resistance towards multiple pathogens is discussed.

Cold water reduces the severity of parasite-inflicted damage: support for wintertime recuperation in aquatic hosts.

The reduction in host fitness caused by parasite infections (virulence) depends on infection intensity and the degree of damage caused per parasite. Environmental conditions can shape both virulence components, but in contrast to infection intensity, environmental impacts on per-parasite damage are poorly understood. Here, we studied the effect of ambient temperature on per-parasite damage, which is jointly determined by the ability of parasites to induce harm (per-parasite pathogenicity) and the ability of hosts to limit damage (tolerance). We experimentally exposed two salmonid species, Atlantic salmon (Salmo salar) and sea trout (Salmo trutta), to replicated genotypes of the eye fluke Diplostomum pseudospathaceum. After development of health damage (eye cataracts) in warm water (16°C) during the first 12weeks post exposure, we maintained the fish at either 5°C (cold water) or 16°C for another 8weeks and quantified changes in cataracts as a function of parasite load. We found that per-parasite damage was reduced in cold compared to warm water, suggesting that cold temperatures improved host health. Per-parasite damage was also affected by parasite genotype and host species, but these effects did not change with temperature. Our findings suggest that cold-water seasons, which are often neglected in host-parasite studies due to low infection risk, could allow hosts to recuperate and thus, may have important implications for the ecology and epidemiology of parasite infections.

Negative associations between parasite avoidance, resistance and tolerance predict host health in salmonid fish populations.

Genetic variation in defence against parasite infections is fundamental for host-parasite evolution. The overall level of defence of a host individual or population includes mechanisms that reduce parasite exposure (avoidance), establishment (resistance) or pathogenicity (tolerance). However, how these traits operate and evolve in concert is not well understood. Here, we investigated genetic variation in and associations between avoidance, resistance and tolerance in a natural host-parasite system. Replicated populations of Atlantic salmon (Salmo salar) and sea trout (an anadromous form of brown trout, Salmo trutta) were raised under common garden conditions and infected with the eye fluke Diplostomum pseudospathaceum. We demonstrate significant genetic variation in the defence traits across host populations and negative associations between the traits, with the most resistant populations showing the weakest avoidance and the lowest infection tolerance. These results are suggestive of trade-offs between different components of defence and possibly underlie the genetic variation in defence traits observed in the wild. Because the three defence mechanisms affect host-parasite evolution in profoundly different ways, we emphasize the importance of studying these traits in concert.

Limited impacts of chronic eye fluke infection on the reproductive success of a fish host

AbstractParasitic infections may affect the reproductive success of the host either directly, through behavioural modification, or indirectly, by altering their reproductive investment in response to infection. We determined the effects of infection with the eye fluke Diplostomum pseudospathaceum (Trematoda) on the reproductive traits of European bitterling (Rhodeus amarus, Cyprinidae), an intermediate fish host with a resource-based mating system. Male bitterling infected by Diplostomum exhibited a larger but less pronounced red eye spot (sexually selected signal) than control males, suggesting that infected males were less preferred by females. The frequency of female ovulation and number of offspring were comparable between the infected and the control group, although there was a 1–2 week delay in the peak of ovulation and offspring production in infected fish, which is known to coincide with higher juvenile mortality. Chronic eye fluke infection had minimal metabolic costs (measured as oxygen consumption) and, consistent with these results, reproductive activity did not differ between infected and control fish in an experimental test of intersexual selection. Overall, the impact of eye fluke infection on the reproduction of European bitterling was limited. We consider the potential effect of favourable conditions during experiments (abundant food, access to spawning substrate and lack of predators and co-infections) on experimental outcomes and recognize that the effects of chronic eye fluke infection in natural conditions might be more pronounced.

The right response at the right time: Exploring helminth immune modulation in sticklebacks by experimental coinfection.

Parasites are one of the strongest selective agents in nature. They select for hosts that evolve counter‐adaptive strategies to cope with infection. Helminth parasites are special because they can modulate their hosts’ immune responses. This phenomenon is important in epidemiological contexts, where coinfections may be affected. How different types of hosts and helminths interact with each other is insufficiently investigated. We used the three‐spined stickleback (Gasterosteus aculeatus) – Schistocephalus solidus model to study mechanisms and temporal components of helminth immune modulation. Sticklebacks from two contrasting populations with either high resistance (HR) or low resistance (LR) against S. solidus, were individually exposed to S. solidus strains with characteristically high growth (HG) or low growth (LG) in G. aculeatus. We determined the susceptibility to another parasite, the eye fluke Diplostomum pseudospathaceum, and the expression of 23 key immune genes at three time points after S. solidus infection. D. pseudospathaceum infection rates and the gene expression responses depended on host and S. solidus type and changed over time. Whereas the effect of S. solidus type was not significant after three weeks, T regulatory responses and complement components were upregulated at later time points if hosts were infected with HG S. solidus. HR hosts showed a well orchestrated immune response, which was absent in LR hosts. Our results emphasize the role of regulatory T cells and the timing of specific immune responses during helminth infections. This study elucidates the importance to consider different coevolutionary trajectories and ecologies when studying host‐parasite interactions.

Within-host interactions shape virulence-related traits of trematode genotypes.

Within-host interactions between co-infecting parasites can significantly influence the evolution of key parasite traits, such as virulence (pathogenicity of infection). The type of interaction is expected to predict the direction of selection, with antagonistic interactions favouring more virulent genotypes and synergistic interactions less virulent genotypes. Recently, it has been suggested that virulence can further be affected by the genetic identity of co-infecting partners (G×G interactions), complicating predictions on disease dynamics. Here, we used a natural host-parasite system including a fish host and a trematode parasite to study the effects of G×G interactions on infection virulence. We exposed rainbow trout (Oncorhynchus mykiss) either to single genotypes or to mixtures of two genotypes of the eye fluke Diplostomum pseudospathaceum and estimated parasite infectivity (linearly related to pathogenicity of infection, measured as coverage of eye cataracts) and relative cataract coverage (controlled for infectivity). We found that both traits were associated with complex G×G interactions, including both increases and decreases from single infection to co-infection, depending on the genotype combination. In particular, combinations where both genotypes had low average infectivity and relative cataract coverage in single infections benefited from co-infection, while the pattern was opposite for genotypes with higher performance. Together, our results show that infection outcomes vary considerably between single and co-infections and with the genetic identity of the co-infecting parasites. This can result in variation in parasite fitness and consequently impact evolutionary dynamics of host-parasite interactions.

Spatiotemporal and gender-specific parasitism in two species of gobiid fish.

Parasitism is considered a major selective force in natural host populations. Infections can decrease host condition and vigour, and potentially influence, for example, host population dynamics and behavior such as mate choice. We studied parasite infections of two common marine fish species, the sand goby (Pomatoschistus minutus) and the common goby (Pomatoschistus microps), in the brackish water Northern Baltic Sea. We were particularly interested in the occurrence of parasite taxa located in central sensory organs, such as eyes, potentially affecting fish behavior and mate choice. We found that both fish species harbored parasite communities dominated by taxa transmitted to fish through aquatic invertebrates. Infections also showed significant spatiotemporal variation. Trematodes in the eyes were very few in some locations, but infection levels were higher among females than males, suggesting differences in exposure or resistance between the sexes. To test between these hypotheses, we experimentally exposed male and female sand gobies to infection with the eye fluke Diplostomum pseudospathaceum. These trials showed that the fish became readily infected and females had higher parasite numbers, supporting higher susceptibility of females. Eye fluke infections also caused high cataract intensities among the fish in the wild. Our results demonstrate the potential of these parasites to influence host condition and visual abilities, which may have significant implications for survival and mate choice in goby populations.

Immunity comes first: The effect of parasite genotypes on adaptive immunity and immunization in three-spined sticklebacks

Adaptive immunity in vertebrates can confer increased resistance against invading pathogens upon re-infection. But how specific parasite genotypes affect the temporal transition from innate to adaptive immunity under continual exposure to parasites is poorly understood. Here, we investigated the effects of homologous and heterologous exposures of genetically distinct parasite lineages of the eye fluke Diplostomum pseudospathaceum on gene expression patterns of adaptive immunity in sticklebacks (Gasterosteus aculeatus). Observable differences in gene expression were largely attributable to final exposures while there was no transcription pattern characteristic for a general response to repeated infections with D. pseudospathaceum. None of the final exposure treatments was able to erase the distinct expression patterns resulting from a heterologous pre-exposed fish. Interestingly, heterologous final exposures showed similarities between different treatment groups subjected to homologous pre-exposure. The observed pattern was supported by parasite infection rates and suggests that host immunization was optimized towards an adaptive immune response that favored effectiveness against parasite diversity over specificity.

Experimental evidence for parasite-induced over-winter mortality in juvenile Rhodeus amarus.

In this study, the effects of the eye fluke Diplostomum pseudospathaceum (Trematoda) infection on over-winter survival of young-of-the-year (YOY) European bitterling Rhodeus amarus (Cyprinidae) were examined between September 2010 and April 2011. The fish were reared in semi-natural conditions to ensure that results were not confounded by other parasite infections. The cumulative mortality of R. amarus from November until April was significantly higher in D. pseudospathaceum-infected fish (57·3%) compared to controls (42·1%). Infection of the parental generation did not have any effect on the mortality of juveniles. The results indicate that D. pseudospathaceum infection increases over-winter mortality of YOY R. amarus. The possible mechanisms causing mortality are discussed.

Differences in susceptibility and immune responses of three-spined sticklebacks (Gasterosteus aculeatus) from lake and river ecotypes to sequential infections with the eye fluke Diplostomum pseudospathaceum.

BackgroundThe eye fluke Diplostomum pseudospathaceum is a frequent parasite of many fresh-water fish species, among those three-spined sticklebacks, particularly in lakes with lymnaeid snails, its first intermediate hosts. Cercariae released from host-snails, penetrate the skin of their fish hosts and within 24 h migrate to the immunologically inert eye lenses. Thus, individual D. pseudospathaceum are exposed to the fish immune system only for a short time, suggesting that only innate immunity can be active against the parasite. However, in nature sticklebacks are exposed to D. pseudospathaceum repeatedly since snails are shedding cercariae from late spring to autumn. Therefore, acquired immunity after initial infection would be advantageous against subsequent parasite encounters.MethodsWe investigated if sticklebacks originating from a lake with high and from a river with low prevalence of D. pseudospathaceum differ in susceptibility to repeated exposure to the parasite. We compared infection success and immune functions in laboratory-bred sticklebacks from both habitats in naïve fish with fish that had been pre-exposed to eye flukes. Head kidney leukocytes (HKL) from experimental sticklebacks were investigated for respiratory burst activity and the proliferation of lymphocytes and monocytes 1.5, 5 and 15 days after infection.ResultsLake sticklebacks were less susceptible than river sticklebacks, however, in both populations pre-exposure led to a similar relative reduction in infection success. The respiratory burst activity was higher with HKL from lake sticklebacks and was up-regulated in pre-exposed fish but dropped 1.5d after an additional exposure, suggesting that activation of phagocytic cells is crucial for the defense against D. pseudospathaceum. Changes in lymphocyte proliferation were only detectable 1.5d after the last exposure in lake sticklebacks, but not 5 and 15d post exposure, indicating that a lymphocyte mediated acquired immune response was not induced. Proliferation of monocytes was significantly increased 1.5d after the last exposure with HKL from both stickleback populations.ConclusionsIncreased resistance to D. pseudospathaceum in sticklebacks from both populations upon pre-exposure cannot be explained by a prominent adaptive immune response. Monocytic leukocytes were more responsive, suggesting that rather cells of the innate than the adaptive immune system are active in the defense of D. pseudospathaceum.

Genotype-Specific vs. Cross-Reactive Host Immunity against a Macroparasite

Vertebrate hosts often defend themselves against several co-infecting parasite genotypes simultaneously. This has important implications for the ecological dynamics and the evolution of host defence systems and parasite strategies. For example, it can drive the specificity of the adaptive immune system towards high genotype-specificity or cross-reactivity against several parasite genotypes depending on the sequence and probability of re-infections. However, to date, there is very little evidence on these interactions outside mammalian disease literature. In this study we asked whether genotype-specific or cross-reactive responses dominate in the adaptive immune system of a fish host towards a common macroparasite. In other words, we investigated if the infection success of a parasite genotype is influenced by the immunization genotype. We reciprocally immunized and re-exposed rainbow trout (Oncorhynchus mykiss) to a range of genotypes of the trematode eye fluke Diplostomum pseudospathaceum, and measured infection success of the parasite. We found that the infection success of the parasite genotypes in the re-exposure did not depend on the immunization genotype. While immunization reduced average infection success by 31%, the reduction was not larger against the initial immunization genotype. Our results suggest significant cross-reactivity, which may be advantageous for the host in genetically diverse re-exposures and have significant evolutionary implications for parasite strategies. Overall, our study is among the first to demonstrate cross-reactivity of adaptive immunity against genetically diverse macroparasites with complex life cycles.

Local differences in immunocompetence reflect resistance of sticklebacks against the eye fluke Diplostomum pseudospathaceum

We investigated population differences in immunological adaptation of three-spined sticklebacks (Gasterosteus aculeatus) to one of their most abundant macroparasites, the eye fluke Diplostomum pseudospathaceum. We compared infection success in lab-bred fish of 2 populations in northern Germany, from a lake, where eye flukes are prevalent, and a river, where these parasites do not occur. In order to discriminate between protection through innate and acquired immunity, we exposed fish either only once or repeatedly. Lake fish were significantly less susceptible than river sticklebacks already after a single exposure, indicating that in sympatric hosts innate immunity plays the major role in the defence against this helminth infection. In both habitat types, previous exposures only marginally decreased infection rates within 12 weeks. Lake fish showed higher immunocompentence by means of respiratory burst activity and spleen size, regardless of the infection status. Furthermore, they were in a better energy status than river fish, as indicated by a higher hepatosomatic index and haematocrit value. Interestingly, F1 hybrid fish of both populations ranged between the pure habitat types in parasite susceptibility as well as in immunological and condition parameters. Our results suggest that sticklebacks from lakes are better adapted to cope with higher parasite abundance in this habitat.

Polymorphic microsatellite loci for the trematode Diplostomum pseudospathaceum

AbstractWe present primers for five polymorphic microsatellite loci in the eye fluke Diplostomum pseudospathaceum (Trematoda), a widely distributed parasite with a complex life cycle used as a model for parasitology and fish immunology. The loci were identified using a GA/CT‐enriched genomic library by subtractive hybridization with magnetic particles. All five loci were highly polymorphic, displaying 17 to 61 alleles and heterozygosities ranging from 0.53 to 0.92. We isolated populations of parasites within the first (snail) and second (fish) intermediate host and found small but significant genetic differentiation (FST = 0.012) between the two life stages of the parasite.