Parasite Release Research Articles

A malaria parasite subtilisin propeptide-like protein is a potent inhibitor of the egress protease SUB1.

Subtilisin-like serine peptidases (subtilases) play important roles in the life cycle of many organisms, including the protozoan parasites that are the causative agent of malaria, Plasmodium spp. As with other peptidases, subtilase proteolytic activity has to be tightly regulated in order to prevent potentially deleterious uncontrolled protein degradation. Maturation of most subtilases requires the presence of an N-terminal propeptide that facilitates folding of the catalytic domain. Following its proteolytic cleavage, the propeptide acts as a transient, tightly bound inhibitor until its eventual complete removal to generate active protease. Here we report the identification of a stand-alone malaria parasite propeptide-like protein, called SUB1-ProM, encoded by a conserved gene that lies in a highly syntenic locus adjacent to three of the four subtilisin-like genes in the Plasmodium genome. Template-based modelling and ab initio structure prediction showed that the SUB1-ProM core structure is most similar to the X-ray crystal structure of the propeptide of SUB1, an essential parasite subtilase that is discharged into the parasitophorous vacuole (PV) to trigger parasite release (egress) from infected host cells. Recombinant Plasmodium falciparum SUB1-ProM was found to be a fast-binding, potent inhibitor of P. falciparum SUB1, but not of the only other essential blood-stage parasite subtilase, SUB2, or of other proteases examined. Mass-spectrometry and immunofluorescence showed that SUB1-ProM is expressed in the PV of blood stage P. falciparum, where it may act as an endogenous inhibitor to regulate SUB1 activity in the parasite.

Exosome Biogenesis in the Protozoa Parasite Giardia lamblia: A Model of Reduced Interorganellar Crosstalk.

Extracellular vesicles (EVs) facilitate intercellular communication and are considered a promising therapeutic tool for the treatment of infectious diseases. These vesicles involve microvesicles (MVs) and exosomes and selectively transfer proteins, lipids, mRNAs, and microRNAs from one cell to another. While MVs are formed by extrusion of the plasma membrane, exosomes are a population of vesicles of endosomal origin that are stored inside the multivesicular bodies (MVBs) as intraluminal vesicles (ILVs) and are released when the MVBs fuse with the plasma membrane. Biogenesis of exosomes may be driven by the endosomal sorting complex required for transport (ESCRT) machinery or may be ESCRT independent, and it is still debated whether these are entirely separate pathways. In this manuscript, we report that the protozoan parasite, Giardia lamblia, although lacking a classical endo-lysosomal pathway, is able to produce and release exosome-like vesicles (ElV). By using a combination of biochemical and cell biology analyses, we found that the ElVs have the same size, shape, and protein and lipid composition as exosomes described for other eukaryotic cells. Moreover, we established that some endosome/lysosome peripheral vacuoles (PVs) contain ILV during the stationary phase. Our results indicate that ILV formation and ElV release depend on the ESCRT-associated AAA+-ATPase Vps4a, Rab11, and ceramide in this parasite. Interestingly, EIV biogenesis and release seems to occur in Giardia despite the fact that this parasite has lost most of the ESCRT machinery components during evolution and is unable to produce ceramide de novo. The differences in protozoa parasite EV composition, origin, and release may reveal functional and structural properties of EVs and, thus, may provide information on cell-to-cell communication and on survival mechanisms.

Rapid transmission of Bonamia exitiosa by cohabitation causes mortality in Ostrea angasi.

The haplosporidian Bonamia was first detected in Australian shellfish in 1991. Australian isolates in Ostrea angasi Sowerby,1871 were identified as Bonamia exitiosa Hine, Cochennac and Berthe, 2001, which threatens development of an O. angasi aquaculture industry. European field data suggest that Bonamia ostreae Pichot, Comps, Tigé, Grizel and Rabouin,1980 infections in Ostreaedulis Linnaeus, 1758 build slowly, but infection dynamics of B.exitiosa in O.angasi are unknown. We investigated B. exitiosa infection in O.angasi by cohabiting uninfected juvenile O.angasi with adults infected with B.exitiosa. Oysters were sampled at 10, 21 and 40days after cohabitation, and B. exitiosa prevalence and intensity were assessed. Bonamia exitiosa rapidly infected and caused disease in O.angasi. Mortalities began at 12days, with ˜50% mortality by day 21 and >85% mortality by day 40. Mortalities displayed pathology consistent with clinical B. exitiosa infection. Time to first infection is likely influenced by a combination of parasite infectivity, host exposure and host immune capacity. Host death is not required for transmission, but probably facilitates release of parasites from decaying tissue. Understanding B. exitiosa transmission informs design and interpretation of field studies and aids development of management strategies for oyster aquaculture.

Actions of Angeli's salt, a nitroxyl (HNO) donor, on Trypanosoma cruzi infection

Chagas disease, caused by Trypanosoma cruzi infection, is endemic to much of Latin America, but also present in the United States (U.S.) The goal of treatment is to eliminate the parasite and decrease the probability of cardiomyopathy and interrupt the cycle of disease transmission. Currently, only benznidazole (BZN) and nifurtimox are recognized by the World Health Organization as effective drugs for treatment of Chagas disease, but both present low cure rates in the chronic phase and often have serious side effects. T. cruzi infection produces an intense inflammatory response in several tissues which is critical for controlling parasites proliferation and evolution of Chagas disease. Compounds liberating nitric oxide (NO donors) have been used as therapeutic agents against T. cruzi. Here we test HNO donor, Angeli's salt (AS) in the outcome of T. cruzi infection. C57BL/6 mice were infected with T. cruzi (Y strain, 5 × 103 trypomastigotes) intraperitoneally (ip). After 15 minutes of infection and the subsequent 12 days, the animals were treated with different concentrations of AS ip. The control group received 100 uL of PBS by the same route. Treatment with AS showed decreased parasitemia and in the number of nests in the cardiac tissue, decreased oxidative stress in erythrocytes and improved leukopenia and thrombocytopenia resulting in reduced disease severity. The in vitro treatment was able to reduce the parasite internalization rate and release of infective trypomastigotes by these cells. Our results suggest for the first time the therapeutic potential of AS in a mouse model of Chagas disease.Support or Funding InformationCAPES, Fundação Araucária, CNPQ, UniZambeze and MCTESTP Mozambique.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Cytomegalovirus vectors expressing Plasmodium knowlesi antigens induce immune responses that delay parasitemia upon sporozoite challenge.

The development of a sterilizing vaccine against malaria remains one of the highest priorities for global health research. While sporozoite vaccines targeting the pre-erythrocytic stage show great promise, it has not been possible to maintain efficacy long-term, likely due to an inability of these vaccines to maintain effector memory T cell responses in the liver. Vaccines based on human cytomegalovirus (HCMV) might overcome this limitation since vectors based on rhesus CMV (RhCMV), the homologous virus in rhesus macaques (RM), elicit and indefinitely maintain high frequency, non-exhausted effector memory T cells in extralymphoid tissues, including the liver. Moreover, RhCMV strain 68–1 elicits CD8+ T cells broadly recognizing unconventional epitopes exclusively restricted by MHC-II and MHC-E. To evaluate the potential of these unique immune responses to protect against malaria, we expressed four Plasmodium knowlesi (Pk) antigens (CSP, AMA1, SSP2/TRAP, MSP1c) in RhCMV 68–1 or in Rh189-deleted 68–1, which additionally elicits canonical MHC-Ia-restricted CD8+ T cells. Upon inoculation of RM with either of these Pk Ag expressing RhCMV vaccines, we obtained T cell responses to each of the four Pk antigens. Upon challenge with Pk sporozoites we observed a delayed appearance of blood stage parasites in vaccinated RM consistent with a 75–80% reduction of parasite release from the liver. Moreover, the Rh189-deleted RhCMV/Pk vectors elicited sterile protection in one RM. Once in the blood, parasite growth was not affected. In contrast to T cell responses induced by Pk infection, RhCMV vectors maintained sustained T cell responses to all four malaria antigens in the liver post-challenge. The delayed appearance of blood stage parasites is thus likely due to a T cell-mediated inhibition of liver stage parasite development. As such, this vaccine approach can be used to efficiently test new T cell antigens, improve current vaccines targeting the liver stage and complement vaccines targeting erythrocytic antigens.

Lessepsian migration and parasitism: richness, prevalence and intensity of parasites in the invasive fishSphyraena chrysotaeniacompared to its native congenerSphyraena sphyraenain Tunisian coastal waters

BackgroundParasites can play various roles in the invasion of non-native species, but these are still understudied in marine ecosystems. This also applies to invasions from the Red Sea to the Mediterranean Sea via the Suez Canal, the so-called Lessepsian migration. In this study, we investigated the role of parasites in the invasion of the Lessepsian migrant Sphyraena chrysotaenia in the Tunisian Mediterranean Sea.MethodsWe compared metazoan parasite richness, prevalence and intensity of S. chrysotaenia (Perciformes: Sphyraenidae) with infections in its native congener Sphyraena sphyraena by sampling these fish species at seven locations along the Tunisian coast. Additionally, we reviewed the literature to identify native and invasive parasite species recorded in these two hosts.ResultsOur results suggest the loss of at least two parasite species of the invasive fish. At the same time, the Lessepsian migrant has co-introduced three parasite species during the initial migration to the Mediterranean Sea, that are assumed to originate from the Red Sea of which only one parasite species has been reported during the spread to Tunisian waters. In addition, we found that the invasive fish has acquired six parasite species that are native in the Mediterranean Sea. However, parasite richness, prevalence and intensity were overall much lower in the invasive compared to the native fish host in the Mediterranean Sea.DiscussionThese results suggest that the Lessepsian migrant may affect native fish hosts by potentially altering the dynamics of native and invasive parasite-host interactions via parasite release, parasite co-introduction and parasite acquisition. They further suggest that the lower infection levels in the invasive fish may result in a competitive advantage over native fish hosts (enemy release hypothesis). This study demonstrates that cross-species comparisons of parasite infection levels are a valuable tool to identify the different roles of parasites in the course of Lessepsian migrations.

Extracellular Vesicles Shed By Trypanosoma cruzi Potentiate Infection and Elicit Lipid Body Formation and PGE2 Production in Murine Macrophages.

During the onset of Trypanosoma cruzi infection, an effective immune response is necessary to control parasite replication and ensure host survival. Macrophages have a central role in innate immunity, acting as an important trypanocidal cell and triggering the adaptive immune response through antigen presentation and cytokine production. However, T. cruzi displays immune evasion mechanisms that allow infection and replication in macrophages, favoring its chronic persistence. One potential mechanism is the release of T. cruzi strain Y extracellular vesicle (EV Y), which participate in intracellular communication by carrying functional molecules that signal host cells and can modulate the immune response. The present work aimed to evaluate immune modulation by EV Y in C57BL/6 mice, a prototype resistant to infection by T. cruzi strain Y, and the effects of direct EV Y stimulation of macrophages in vitro. EV Y inoculation in mice prior to T. cruzi infection resulted in increased parasitemia, elevated cardiac parasitism, decreased plasma nitric oxide (NO), reduced NO production by spleen cells, and modulation of cytokine production, with a reduction in TNF-α in plasma and decreased production of TNF-α and IL-6 by spleen cells from infected animals. In vitro assays using bone marrow-derived macrophages showed that stimulation with EV Y prior to infection by T. cruzi increased the parasite internalization rate and release of infective trypomastigotes by these cells. In this same scenario, EV Y induced lipid body formation and prostaglandin E2 (PGE2) production by macrophages even in the absence of T. cruzi. In infected macrophages, EV Y decreased production of PGE2 and cytokines TNF-α and IL-6 24 h after infection. These results suggest that EV Y modulates the host response in favor of the parasite and indicates a role for lipid bodies and PGE2 in immune modulation exerted by EVs.

The impact of parasites during range expansion of an invasive gecko.

Host-parasite dynamics can play a fundamental role in both the establishment success of invasive species and their impact on native wildlife. The net impact of parasites depends on their capacity to switch effectively between native and invasive hosts. Here we explore host-switching, spatial patterns and simple fitness measures in a slow-expanding invasion: the invasion of Asian house geckos (Hemidactylus frenatus) from urban areas into bushland in Northeast Australia. In bushland close to urban edges, H. frenatus co-occurs with, and at many sites now greatly out-numbers, native geckos. We measured prevalence and intensity of Geckobia mites (introduced with H. frenatus), and Waddycephalus (a native pentastome). We recorded a new invasive mite species, and several new host associations for native mites and geckos, but we found no evidence of mite transmission between native and invasive geckos. In contrast, native Waddycephalus nymphs were commonly present in H. frenatus, demonstrating this parasite's capacity to utilize H. frenatus as a novel host. Prevalence of mites on H. frenatus decreased with distance from the urban edge, suggesting parasite release towards the invasion front; however, we found no evidence that mites affect H. frenatus body condition or lifespan. Waddycephalus was present at low prevalence in bushland sites and, although its presence did not affect host body condition, our data suggest that it may reduce host survival. The high relative density of H. frenatus at our sites, and their capacity to harbour Waddycephalus, suggests that there may be impacts on native geckos and snakes through parasite spillback.

Enhanced fecundity and parasite release in the first amphipod invader on the Iberian Peninsula

Crangonyx pseudogracilis is a North American amphipod recently detected in Portugal and it is the first invasive freshwater amphipod detected in the Iberian Peninsula. The aim of this study was to assess the population structure and its invasion range in this area, but also to assess its pathogen profile and its origin through genetic analyses. Crangonyx pseudogracilis was only found in the lowlands of the Tagus River basin, a restricted area of the three sampled river basins (Tagus, Guadiana and Sado). Only one mitochondrial DNA (mtDNA) gene cytochrome oxidase subunit 1 (COI) haplotype, identical to the one already detected in Europe and in the Lake Charles, USA, was identified. The obtained data show that the studied populations are well-established, have a high female fecundity and a reproduction period extended throughout the year. No parasites were detected in the 236 dissected specimens. The dissimilarity of parasites prevalence to other European invasive populations opens the possibility of a direct introduction from North America but it also does not exclude the possibility of a secondary introduction from another European country.

Circadian rhythms of trematode parasites: applying mixed models to test underlying patterns.

Circadian rhythms of parasites and their hosts can influence processes such as transmission, pathology and life cycle evolution. For trematode parasites that depend on free-living infectious stages (i.e. cercariae) to move among host species, the timing of parasite release is hypothesized to increase the likelihood of contacting a host. Yet, a persistent challenge in studying such biorhythms involves selection of appropriate analytical techniques. Here, we extend a generalized linear mixed modelling (GLMM) framework to cosinor analyses, thereby allowing flexibility in the statistical distribution of the response variable, incorporation of multiple covariates and inclusion of hierarchical grouping effects. By applying this approach to 93 snails infected with trematode parasites from freshwater pond ecosystems, we detected non-random rhythms in six of eight species, with variation in both the timing of peak cercariae release (between 5:10 and 21:46 h) and its magnitude (between 13 and 386). The use of GLMM yielded more accurate and precise estimates of the cosinor parameters compared with classical least-squares (LS) based on a simulation-based sensitivity analysis. The sensitivity analysis revealed that the amplitude and rhythm-adjusted mean values from the LS models diverged from the true values at some limits. We highlight the importance of novel analytical approaches for evaluating parasite circadian rhythms and investigating their underlying mechanisms.

Liver cystic echinococcosis and human host immune and autoimmune follow-up: A review.

Cystic echinococcosis (CE) is an infectious disease caused by the larvae of parasite Echinococcus granulosus (E. granulosus). To successfully establish an infection, parasite release some substances and molecules that can modulate host immune functions, stimulating a strong anti-inflammatory reaction to carry favor to host and to reserve self-survival in the host. The literature was reviewed using MEDLINE, and an open access search for immunology of hydatidosis was performed. Accumulating data from animal experiments and human studies provided us with exciting insights into the mechanisms involved that affect all parts of immunity. In this review we used the existing scientific data and discuss how these findings assisted with a better understanding of the immunology of E. granulosus infection in man. The aim of this study is to point the several facts that challenge immune and autoimmune responses to protect E. granulosus from elimination and to minimize host severe pathology. Understanding the immune mechanisms of E. granulosus infection in an intermediate human host will provide, we believe, a more useful treatment with immunomodulating molecules and possibly better protection from parasitic infections. Besides that, the diagnosis of CE has improved due to the application of a new molecular tool for parasite identification by using of new recombinant antigens and immunogenic peptides. More studies for the better understanding of the mechanisms of parasite immune evasion is necessary. It will enable a novel approach in protection, detection and improving of the host inflammatory responses. In contrast, according to the “hygiene hypothesis”, clinical applications that decrease the incidence of infection in developed countries and recently in developing countries are at the origin of the increasing incidence of both allergic and autoimmune diseases. Thus, an understanding of the immune mechanisms of E. granulosus infection is extremely important.

Temperature-related parasite infection dynamics: the case of proliferative kidney disease of brown trout.

Climate change, in particular rising temperature, is suspected to be a major driver for the emergence of many wildlife diseases. Proliferative kidney disease of salmonids, caused by the myxozoan Tetracapsuloides bryosalmonae, was used to evaluate how temperature dependence of host-parasite interactions modulates disease emergence. Brown trout (Salmo trutta fario) kept at 12 and 15 °C, were experimentally infected with T. bryosalmonae. Parasite development in the fish host and release of spores were quantified simultaneously to unravel parasite transmission potential from the vertebrate to the invertebrate host. A change to a stable plateau in infection intensity of the kidney coincided with a threshold at which spore shedding commenced. This onset of parasite release was delayed at the low temperature in accordance with reaching this infection intensity threshold, but the amount of spores released was irrespective of temperature. The production of parasite transmission stages declined with time. In conclusion, elevated temperature modifies the parasite transmission opportunities by increasing the duration of transmission stage production, which may affect the spread and establishment of the parasite in a wider range of rivers.

Modeling the transmission of Perkinsus marinus in the Eastern oyster Crassostrea virginica

Dermo disease caused by the protistan Perkinsus marinus in Eastern oysters Crassotrea virginica is an important source of mortality impacting oyster population dynamics resulting in substantial losses in fisheries and aquaculture. The rapid transmission and spread of the disease minimized the importance of transmission models and past models (proliferation-based models) assumed simple density-dependent transmission or rapid infection post-settlement. This approach is a good approximation only for low population densities. A transmission model was developed for P. marinus in Eastern oysters that accounts for the seasonal change in disease dynamics and density-dependent foraging (of suspended particles) interference among hosts. The model, verified and evaluated against field observations, incorporates parasite release to the water column from live and dead individuals, parasite consumption by living oysters, the diffusion of parasites in the water, body burden-based dose-dependent transmission, recruitment, and disease-caused mortality. The model returns a basic reproduction number R0 for Dermo much greater than unity (R0=90) in accordance with the current persistence and pandemic nature of this disease in oysters. No population density is obtained that is low enough to suppress R0 below 1 (i.e. disease extinction). R0 is also estimated for high oyster densities (>300 individuals m−2) and particularly for relatively large oysters (∼90mm), today rare but once common before generalized overfishing occurred on healthy oyster reefs. In this scenario, R0 drops below 1, indicating that high oyster density can limit disease invasion through foraging interference and depletion of parasites in the water column. High intensity recruitment events allow the oyster population to attain such densities and limit the development of epizootics. These results provide insight into the transition from past populations, where Dermo is inferred to have been limited in its impact, to the current persistent and pandemic nature of this disease. Further coupling of this model into metapopulation and hydrodynamic models could be a promising tool to support management decision-making for bivalve populations impacted by Dermo disease.

Quantitative analysis of Cryptosporidium growth in in vitro culture--the impact of parasite density on the success of infection.

Cryptosporidium is an important waterborne pathogen for which no treatment or vaccination is available. This study set out to quantify DNA replication of Cryptosporidium parvum in vitro. Cryptosporidium DNA could be detected at up to 60 % of input level in both host-cell-free and host cell containing cultures 6 days after infection with living sporozoites, but was lost within 2 days in cultures inoculated with UV-inactivated sporozoites. Total DNA increased between days 2 and 6, evidence of successful DNA replication in both cell-free and host-cell-containing cultures. Overall however, only a small fraction (up to 5 %) of parasite DNA could be found associated with host cells or bound to plastic of the cell-free cultures, and the majority of parasite DNA was present in the cell culture medium, separable by simple decantation. After 2 days, in host-cell-containing cultures, the parasite DNA could be concentrated by slow centrifugation, suggesting that it was associated with intact parasite cells, but at 6 days, the majority could not be centrifuged and is therefore thought to have represented copies associated with dead and degraded parasites. In cell-free cultures and in larger plates, the majority of DNA was in this form. Performance of the parasite was best in small culture plates, and least in the largest plate sizes. We interpret these results as suggesting that Cryptosporidium sporozoites first bind to the host cell monolayer or to the plasticware, but then by 2 days, there has been a substantial release of parasites back into the medium. Host-cell-free cultures also supported modest replication and may have represented DNA synthesis in cells beginning merogony. The role of the host cells is unclear, as so much of the parasite DNA is released into the medium. Host cells may provide a feeder role, conditioning the medium for Cryptosporidium development.

Parting ways: parasite release in nature leads to sex-specific evolution of defence.

We evaluated the extent to which males and females evolve along similar or different trajectories in response to the same environmental shift. Specifically, we used replicate experimental introductions in nature to consider how release from a key parasite (Gyrodactylus) generates similar or different defence evolution in male vs. female guppies (Poecilia reticulata). After 4-8 generations of evolution, guppies were collected from the ancestral (parasite still present) and derived (parasite now absent) populations and bred for two generations in the laboratory to control for nongenetic effects. These F2 guppies were then individually infected with Gyrodactylus, and infection dynamics were monitored on each fish. We found that parasite release in nature led to sex-specific evolutionary responses: males did not show much evolution of resistance, whereas females showed the evolution of increased resistance. Given that male guppies in the ancestral population had greater resistance to Gyrodactylus than did females, evolution in the derived populations led to reduction of sexual dimorphism in resistance. We argue that previous selection for high resistance in males constrained (relative to females) further evolution of the trait. We advocate more experiments considering sex-specific evolutionary responses to environmental change.

Global parasite and Rattus rodent invasions: The consequences for rodent-borne diseases.

We summarize the current knowledge on parasitism-related invasion processes of the globally invasive Rattus lineages, originating from Asia, and how these invasions have impacted the local epidemiology of rodent-borne diseases. Parasites play an important role in the invasion processes and successes of their hosts through multiple biological mechanisms such as "parasite release," "immunocompetence advantage," "biotic resistance" and "novel weapon." Parasites may also greatly increase the impact of invasions by spillover of parasites and other pathogens, introduced with invasive hosts, into new hosts, potentially leading to novel emerging diseases. Another potential impact is the ability of the invader to amplify local parasites by spillback. In both cases, local fauna and humans may be exposed to new health risks, which may decrease biodiversity and potentially cause increases in human morbidity and mortality. Here we review the current knowledge on these processes and propose some research priorities.

Host ER stress during malaria parasite infection

After transmission by Anopheles mosquitoes, malaria parasite sporozoites target the liver, where they infect hepatocytes and multiply thousands of times. The release of new parasites into the blood stream then initiates symptomatic red blood cell infection. Although successful replication within hepatocytes is critical for host infection, little is known about parasite–hepatocyte interactions that ensure parasite survival and development. In this issue of EMBO Reports , the Mota group describes a beneficial role of the host ER stress pathway for Plasmodium survival in infected hepatocytes [[1][1]]. They demonstrate that proteins and transcripts that act in the unfolded protein response (UPR) are elevated in hepatocytes in response to infection. Reversing these perturbations by eliminating the splicing of XBP1 or knockdown of CREBH is detrimental to parasite development. These findings are of significant interest in light of recent findings that elucidate other aspects of liver‐stage parasite–hepatocyte interactions and raise new, intriguing questions for the field (Fig [1][2]). [1]: #ref-1 [2]: #F1

Tetracapsuloides bryosalmonae infection affects the expression of genes involved in cellular signal transduction and iron metabolism in the kidney of the brown trout Salmo trutta.

Tetracapsuloides bryosalmonae is an enigmatic endoparasite which causes proliferative kidney disease in various species of salmonids in Europe and North America. The life cycle of the European strain of T. bryosalmonae generally completes in an invertebrate host freshwater bryozoan and vertebrate host brown trout (Salmo trutta) Linnaeus, 1758. Little is known about the gene expression in the kidney of brown trout during the developmental stages of T. bryosalmonae. In the present study, quantitative real-time PCR was applied to quantify the target genes of interest in the kidney of brown trout at different time points of T. bryosalmonae development. PCR primers specific for target genes were designed and optimized, and their gene expression levels were quantified in the cDNA kidney samples using SYBR Green Supermix. Expression of Rab GDP dissociation inhibitor beta, integral membrane protein 2B, NADH dehydrogenase 1 beta subcomplex subunit 6, and 26S protease regulatory subunit S10B were upregulated significantly in infected brown trout, while the expression of the ferritin M middle subunit was downregulated significantly. These results suggest that host genes involved in cellular signal transduction, proteasomal activities, including membrane transporters and cellular iron storage, are differentially upregulated or downregulated in the kidney of brown trout during parasite development. The gene expression pattern of infected renal tissue may support the development of intraluminal sporogonic stages of T. bryosalmonae in the renal tubular lumen of brown trout which may facilitate the release of viable parasite spores to transmit to the invertebrate host bryozoan.

A Plasmodium phospholipase is involved in disruption of the liver stage parasitophorous vacuole membrane.

The coordinated exit of intracellular pathogens from host cells is a process critical to the success and spread of an infection. While phospholipases have been shown to play important roles in bacteria host cell egress and virulence, their role in the release of intracellular eukaryotic parasites is largely unknown. We examined a malaria parasite protein with phospholipase activity and found it to be involved in hepatocyte egress. In hepatocytes, Plasmodium parasites are surrounded by a parasitophorous vacuole membrane (PVM), which must be disrupted before parasites are released into the blood. However, on a molecular basis, little is known about how the PVM is ruptured. We show that Plasmodium berghei phospholipase, PbPL, localizes to the PVM in infected hepatocytes. We provide evidence that parasites lacking PbPL undergo completely normal liver stage development until merozoites are produced but have a defect in egress from host hepatocytes. To investigate this further, we established a live-cell imaging-based assay, which enabled us to study the temporal dynamics of PVM rupture on a quantitative basis. Using this assay we could show that PbPL-deficient parasites exhibit impaired PVM rupture, resulting in delayed parasite egress. A wild-type phenotype could be re-established by gene complementation, demonstrating the specificity of the PbPL deletion phenotype. In conclusion, we have identified for the first time a Plasmodium phospholipase that is important for PVM rupture and in turn for parasite exit from the infected hepatocyte and therefore established a key role of a parasite phospholipase in egress.

How temperature shifts affect parasite production: testing the roles of thermal stress and acclimation

Summary Changes in the magnitude and frequency of temperature shifts with climate change will influence species interactions if species have differential acclimation responses. For example, if parasites acclimate to temperature shifts faster than their hosts, as might be expected due to their smaller sizes and faster metabolisms, temperature variability could lead to increased infection. However, this assumption might not hold if benefits of acclimation are counteracted by energetic costs or thermal stress, underscoring the need for empirical efforts to assess how temperature variability will influence host–parasite interactions. We used an array of replicate incubators to test how temperature shifts from five acclimation temperatures (13–25 °C) to five performance temperatures (16–28 °C) influenced release of infective stages by the trematode parasite Ribeiroia ondatrae from its snail intermediate host (Helisoma trivolvis) at four time‐points after a temperature shift. Initially, parasite release was higher at warm temperatures and increased temporarily after infected snails were shifted to higher temperatures, particularly for hosts acclimated to cooler temperatures. However, these effects were transient, such that parasite release at warm temperatures declined steadily over the 7 days following the shift. Warmer temperatures also increased snail mortality. Parasite release was strongly influenced not only by ambient temperature but also by the thermal history of the host. Prior acclimation to warm temperatures reduced parasite release at warm performance temperatures, contrary to the beneficial acclimation hypothesis. Rather, the observed pattern was likely driven by: (i) energetic costs of prolonged exposure to high temperatures (‘thermal stress’) or (ii) parasites' capacity to ‘store’ infectious stages at cooler temperatures. The time‐dependent nature of thermal effects on parasite performance highlights the importance of considering the amplitude and frequency of temperature variability for understanding future changes to disease dynamics.