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

BACKGROUND: The 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. METHODS: We 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. RESULTS: Lake 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. CONCLUSIONS: Increased 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

Characterization of immune responses of carp (Cyprinus carpio L.) to the hemoflagellate Trypanoplasma borreli Laveran and Mesnil, 1901

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Induction of diploid gynogenesis in an evolutionary model organism, the three-spined stickleback (Gasterosteus aculeatus)

Background: Rapid advances in genomics have provided nearly complete genome sequences for many different species. However, no matter how the sequencing technology has improved, natural genetic polymorphism complicates the production of high quality reference genomes. To address this problem, researchers have tried using artificial modes of genome manipulation such as gynogenesis for fast production of inbred lines. Results: Here, we present the first successful induction of diploid gynogenesis in an evolutionary model system, the three-spined sticklebacks (Gasterosteus aculeatus), using a combination of UV-irradiation of the sperm and heat shock (HS) of the resulting embryo to inhibit the second meiotic division. Optimal UV irradiation of the sperm was established by exposing stickleback sperm to a UV- light source at various times. Heat shock parameters like temperature, duration, and time of initiation were tested by subjecting eggs fertilized with UV inactivated sperm 5, 10, 15, 20, 25, or 30 minutes post fertilization (mpf) to 30°C, 34°C, or 38°C for 2, 4, 6 or 8 minutes. Gynogen yield was highest when stickleback eggs were activated with 2 minutes UV-irradiated sperm and received HS 5 mpf at 34°C for 4 minutes. Conclusions: Diploid gynogenesis has been successfully performed in three-spined stickleback. This has been confirmed by microsatellite DNA analysis which revealed exclusively maternal inheritance in all gynogenetic fry tested. Ploidy verification by flow cytometry showed that gynogenetic embryos/larvae exhibiting abnormalities were haploids and those that developed normally were diploids, i.e., double haploids that can be raised until adult size

Environmental temperature variation influences fitness trade-offs and tolerance in a fish-tapeworm association

Background Increasing temperatures are predicted to strongly impact host-parasite interactions, but empirical tests are rare. Host species that are naturally exposed to a broad temperature spectrum offer the possibility to investigate the effects of elevated temperatures on hosts and parasites. Using three-spined sticklebacks, Gasterosteus aculeatus L., and tapeworms, Schistocephalus solidus (Müller, 1776), originating from a cold and a warm water site of a volcanic lake, we subjected sympatric and allopatric host-parasite combinations to cold and warm conditions in a fully crossed design. We predicted that warm temperatures would promote the development of the parasites, while the hosts might benefit from cooler temperatures. We further expected adaptations to the local temperature and mutual adaptations of local host-parasite pairs. Results Overall, S. solidus parasites grew faster at warm temperatures and stickleback hosts at cold temperatures. On a finer scale, we observed that parasites were able to exploit their hosts more efficiently at the parasite’s temperature of origin. In contrast, host tolerance towards parasite infection was higher when sticklebacks were infected with parasites at the parasite’s ‘foreign’ temperature. Cold-origin sticklebacks tended to grow faster and parasite infection induced a stronger immune response. Conclusions Our results suggest that increasing environmental temperatures promote the parasite rather than the host and that host tolerance is dependent on the interaction between parasite infection and temperature. Sticklebacks might use tolerance mechanisms towards parasite infection in combination with their high plasticity towards temperature changes to cope with increasing parasite infection pressures and rising temperatures

Comparative transcriptomics of stickleback immune gene responses upon infection by two helminth parasites, Diplostomum pseudospathaceum and Schistocephalus solidus

Immune systems of vertebrates are much more diverse than previously thought, in particular at the base of the vertebrate clade. RNA-seq was used to describe in detail the transcriptomic response of stickleback hosts to infection by two helminth parasites, the trematode . Diplostomum pseudospathaceum (2 genotypes plus a genotype mix) and the cestode . Schistocephalus solidus. Based on a global transcription profiling, we present immune genes that are active during chronic or multiple repeated infection. We found that the transcription profiles of . D. pseudospathaceum genotypes were as divergent as those of the two parasite species. When comparing the host immune response, only 5 immune genes were consistently upregulated upon infection by both species. These genes indicated a role for enhanced toll like receptor (TLR) activity (CTSK, CYP27B1) and an associated positive regulation of macrophages (CYP27B1, THBS1) for general helminth defense. We interpret the largely differentiated gene expression response among parasite species as general redundancy of the vertebrate immune system, which was also visible in genotype-specific responses among the different . D. . pseudospathaceum infections. The present study provides the first evidence that IL4-mediated activation of T-helper lymphocyte cells is also important in anti-helminthic immune responses of teleost fish

Differences on the level of hepatic transcriptome between two flatfish species in response to liver cancer and environmental pollution levels

Environmental factors can cause cancer in both wild animals and humans. In ecological settings, genetic variation and natural selection can sometimes produce resilience to the negative impacts of environmental change. An increase in oncogenic substances in natural habitats has therefore, unintentionally, created opportunities for using polluted habitats to study cancer defence mechanisms. The Baltic and North Sea are among the most contaminated marine areas, with a long history of pollution. Two flatfish species (flounder, Platichthys flesus and dab, Limanda limanda) are used as ecotoxicological indicator species due to pollution-induced liver cancer. Cancer is more prevalent in dab, suggesting species-specific differences in vulnerability and/or defence mechanisms. We conducted gene expression analyses for 30 flatfishes. We characterize between- and within-species patterns in potential cancer-related mechanisms. By comparing cancerous and healthy fishes, and noncancerous fishes from clean and polluted sites, we suggest also genes and related physiological mechanisms that could contribute to a higher resistance to pollution-induced cancer in flounders. We discovered changes in transcriptome related to elevated pollutant metabolism, alongside greater tumour suppression mechanisms in the liver tissue of flounders compared to dabs. This suggests either hormetic upregulation of tumour suppression or a stronger natural selection pressure for higher cancer resistance for flounders in polluted environment. Based on gene expression patterns seen in cancerous and healthy fish, for liver cancer to develop in flounders, genetic defence mechanisms need to be suppressed, while in dabs, analogous process is weak or absent. We conclude that wild species could offer novel insights and ideas for understanding the nature and evolution of natural cancer defence mechanisms.We are grateful to the crew of RW Walther Herwig III for all-round help during the fieldwork. This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 951963.We are grateful to the crew of RW Walther Herwig III for all-round help during the fieldwork. This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 951963

Absence of major histocompatibility complex class II mediated immunity in pipefish, Syngnathus typhle: evidence from deep transcriptome sequencing

The major histocompatibility complex (MHC)-mediated adaptive immune system is the hallmark of gnathostome immune defence. Recent work suggests that cod-like fishes (Gadidae) lack important components of the MHC class II mediated immunity. Here, we report a putative independent loss of functionality of this pathway in another species, the pipefish Syngnathus typhle, that belongs to a distantly related fish family (Syngnathidae). In a deep transcriptome sequencing approach comprising several independent normalized and non-normalized expressed sequence tag (EST) libraries with approximately 7.5 × 108 reads, sequenced with two next generation platforms (454 and Illumina), we were unable to identify MHC class IIα/β genes as well as genes encoding associated receptors. Along with the recent findings in cod, our results suggest that immune systems of the Euteleosts may be more variable than previously assumed

Data from: Consequences of divergent temperature optima in a host-parasite system

It was suggested that parasite infections become more severe with rising temperature, as expected during global warming. In ectothermic systems, the growth of a parasite and therefore its reproductive capacity is expected to increase with temperature. However, the outcome of the interaction depends on the temperature optima of both host and parasite. Here we used experimental infections of three-spined stickleback fish (Gasterosteus aculeatus) with its specific tapeworm parasite Schistocephalus solidus to investigate in detail the temperature optima for both host and parasite. We analyzed the fitness consequences thereof, focusing on growth and immunity of the host, and growth and offspring production of the parasite as fitness correlates. We checked for potential differences among populations, using the offspring of hosts and parasites derived from four study sites in Iceland, Germany and Spain that differ in average annual temperature ranging between 4.8 °C and 18.4 °C. We found differences in temperature optima of host and parasites that were quite consistent across the populations: while sticklebacks grew faster and had higher immune activity at low temperatures, the parasites did not even grow fast enough to reach sexual maturity in these conditions. By contrast, with increasing temperatures, parasite growth, egg production and offspring hatching increased strongly while host immunity and growth were impaired. Our results show that divergent temperature optima of hosts and parasites can have drastic fitness consequences and support the expectation that some parasites will benefit from global warming

Data_Exp_4_Temperature_for_Dryad

Exel file containing data of individual sticklebacks and parasites investigated in the present study. Corresponding columns indicate treatment, origin and tank of each individual

Effects of climate change on marine dumped munitions and possible consequence for inhabiting biota

Marine environments are contaminated with enormous amounts of warfare agents due to military activity and exercise, and the disposal of unused ordnance. Due to corrosion of munition shells, substances are leaking from the warfare materials into the environment. It has to be expected that climate change will influence munition corrosion and distribution of their content. Although there is no doubt about the principle toxicity of many of the munition compounds, including their transformation/degradation products, the impact of munition compounds on marine environments, including their biota are yet only at the beginning to be understood. Recently the intake of munition compounds has been confirmed in mussels and fish collected from contaminated areas. It has become clear that dumped munitions are a continuous source of toxic substances leaking into the environment and that ongoing corrosion will worsen the problem. The present review intends to evaluate the available literature on how climate change might influence the contamination of marine environments and inhabiting biota with munition compounds. Direct testing (or modelling) of climate change scenarios in the context of the marine munition problems has yet not been undertaken. Nevertheless, it can be predicted that climate change effects such as rising temperature and higher frequencies of extreme weather events will accelerate the rates at which disposed ordnance corrodes and consequently accelerate the rate at which munition compounds are leaking out. Climate change will cause elevated stress to biota, ranging from temperature stress and lower availability of oxygen to shifts in salinity and pH. In combination, elevated release of munition related compounds and elevated environmental stress, will put biota under threat, in particular in areas with high munition contamination and limited water exchange, such as the Baltic Sea. On a positive side, biodegradation of organic munition compounds by biota and microorganisms is likely to be accelerated with rising temperature