Overcoming research challenges: In vitro cultivation of Ameson portunus (Phylum Microsporidia).

Ameson portunus is an intracellular pathogen that infects marine crabs Portunus trituberculatus and Scylla paramamosain, causing significant economic losses. However, research into this important parasite has been limited due to the absence of an in vitro culture system. To address this challenge, we developed an in vitro cultivation model of A. portunus using RK13 cell line in this study. The fluorescent labeling assay indicated a high infection rate (∼60 %) on the first day post-infection and quantitative PCR (qPCR) detection demonstrated successful infection as early as six hours post-inoculation. Fluorescence in situ hybridization (FISH) and qPCR were used for the detection of A. portunus infected cells. The FISH probe we designed allowed detection of A. portunus in infected cells and qPCR assay provided accurate quantification of A. portunus in the samples. Transmission electron microscopy (TEM) images revealed that A. portunus could complete its entire life cycle and produce mature spores in RK13 cells. Additionally, we have identified novel life cycle characteristics during the development of A. portunus in RK 13 cells using TEM. These findings contribute to our understanding of new life cycle pathways of A. portunus. The establishment of an in vitro culture model for A. portunus is critical as it provides a valuable tool for understanding the molecular and immunological events that occur during infection. Furthermore, it will facilitate the development of effective treatment strategies for this intracellular pathogen.

Variation in Paramarteilia canceri infections in velvet crab Necora puber.

Sustainable management of crustacean populations requires an understanding of the range of factors affecting different crustacean species. Recently, a high prevalence of a paramyxid parasite, Paramarteilia canceri, was reported in velvet crabs Necora puber in Ireland. Similar parasites have been known to cause mass mortalities in bivalves and, as velvet crabs are an important commercial species, these parasite infections are cause for concern. The main objective of this study was to examine variation in P. canceri infections in relation to host biology and season over a 2 yr period. In addition, we tested a range of host tissues and organs to gain more information on the host-parasite interaction. The parasite was present in all tissues and organs investigated, including the gonad and eggs of a berried female. Parasite prevalence was highest in the cuticular epithelium and hepatopancreas. Both annual and seasonal variation was found in parasite prevalence and parasite load. No difference was found in parasite prevalence or parasite load with either crab size or crab sex. Granulomas as a response to infection were significantly more abundant in infected velvet crab individuals. The results of this study provide important information on the host-parasite interaction between P. canceri and the velvet crab and highlight the importance of including parasite monitoring in the management of crustacean fisheries.

Microbial communities are indicators of parasite infection status.

Growing evidence suggests that microbiomes have been shaping the evolutionary pathways of macroorganisms for millennia and that these tiny symbionts can influence, and possibly even control, species interactions like host-parasite relationships. Yet, while studies have investigated host-parasites and microbiomes separately, little has been done to understand all three groups synergistically. Here, we collected infected and uninfected Eurypanopeus depressus crab hosts from a coastal North Carolina oyster reef three times over 4 months. Infected crabs demonstrated an external stage of the rhizocephalan parasite, Loxothylacus panopaei. Community analyses revealed that microbial richness and diversity were significantly different among tissue types (uninfected crab, infected crab, parasite externae and parasite larvae) and over time (summer and fall). Specifically, the microbial communities from parasite externae and larvae had similar microbiomes that were consistent through time. Infected crabs demonstrated microbial communities spanning those of their host and parasite, while uninfected crabs showed more distinctive communities with greater variability over time. Microbial communities were also found to be indicators of early-stage infections. Resolving the microbial community composition of a host and its parasite is an important step in understanding the microbiome's role in the host-parasite relationship and determining how this tripartite relationship impacts coevolutionary processes.

Sacculina-Induced Morphological Feminization in the Grapsid Crab Pachygrapsus crassipes.

Rhizocephalan barnacles (Thecostraca: Cirripedia) are parasitic crustaceans that lack appendages, segmentation, and a digestive system in adults, while instead infiltrating their hosts with a nutrient-absorbing system of rootlets. Sacculinids, belonging to the Rhizocephala order, are known for their various parasitization-induced effects on their decapod hosts, such as parasitic castration, reduction in the growth of secondary sexual characteristics, feminization of male crabs, and alteration of host behavior. In this study, we conducted field surveys in Japan at Manazuru Town (Kanagawa) on the Pacific coast, and on Sado Island and Noto Peninsula on the Sea of Japan side, and found that sacculinid-parasite-ratios on the grapsid crab Pachygrapsus crassipes were particularly high on the Sea of Japan coast. Molecular phylogenetic analysis revealed that the Manazuru population forms a single clade with Sacculina yatsui, and both Sado and Noto populations form a single clade with S. confragosa. We further demonstrated that external morphologies of male P. crassipes parasitized by sacculinids were changed to female phenotypes. This host-parasite interaction will be a useful model for understanding molecular mechanisms underlying rhizocephalan-driven morphological and behavioral feminization and castration.

Microsporidia Ameson earli sp. n. and A. michaelis (Sprague 1965) infecting blue crabs Callinectes sapidus from shedding facilities in Louisiana.

During a survey for pathogens and commensals of blue crabs in commercial soft shell shedding facilities in Louisiana, we discovered an occurrence of microsporidiosis in two of forty examined crabs. Judging from spore shape and size, tissue tropism and external signs of muscle pathology, the causative agent of infections was identified as Ameson michaelis, a muscle-infecting species that has been repeatedly detected in populations of Callinectes sapidus in Louisiana since 1965. However, retrospective ultrastructural examination revealed that in one of Ameson-infected crabs, infection was caused by a parasite with ultrastructural characters not completely compliant with the ones of A. michaelis. The major difference was the absence of microtubule-like appendages attached to the exospore, typical of A. michaelis and other Ameson spp. SSUrDNA-inferred pairwise evolutionary distances between the novel species and other Ameson spp. ranged from 0.006 to 0.051; it was 0.039 in the case of A. michaelis. Hence, we describe here a new species in the genus Ameson, and name it after Prof. Earl Weidner, our colleague and friend, an outstanding microsporidiologist and the author of pioneer papers on the ultrastructure and physiology of A. michaelis.

Natural transmission of Hematodinium perezi in juvenile blue crabs (Callinectes sapidus) in the laboratory.

Hematodinium perezi is a dinoflagellate endoparasitic in marine crustaceans, primarily decapods. It occurs in juvenile blue crabs, Callinectes sapidus, at high prevalence levels and has severe pathogenic effects in this host. The life history outside the host has not been experimentally investigated and, until now, transmission using dinospores has not been successful. We investigated the natural transmission dynamics of H. perezi in the laboratory using small juvenile crabs, which are highly susceptible to infection in the field, and elevated temperatures, which are known to stimulate dinospore production. Natural water-borne transmission to naïve crabs varied between 7 and 100% and was not correlated with dinospore densities measured from their aquaria water. Infections appeared to develop quickly in naïve hosts at 25 °C, suggesting that elevated temperatures as seen in the late summer and early autumn have a strong influence on the transmission of H. perezi in natural systems.

Comparative genomics for Agmasoma sp. (Microsporidia) parasitising invasive Carcinus aestuarii and Carcinus maenas in Argentina.

Carcinus spp. are global aquatic invaders and carriers of several parasites, including a taxonomically unrecognised microsporidian recently observed from Argentina. We provide genome drafts for two parasite isolates, one from Carcinus maenas and one from Carcinus aestuarii, and use multi-gene phylogenetics and genome comparison methods to outline their similarities. Their SSU genes are 100 % similar and other genes have an average similarity of 99.31 %. We informally name the parasite Agmasoma carcini, terming the isolates Ac. var. aestuarii and Ac. var. maenas, following the wealth of genomic data available for each. This study follows on from Frizzera et al. (2021), where this parasite was first histologically identified.

Pseudohepatospora borealis n. gen. n. sp. (Microsporidia: Enterocytozoonida): A microsporidian pathogen of the Jonah crab (Cancer borealis).

The microsporidian diversity catalogued so far has resulted in the development of several taxonomic groups, one of which is the Enterocytozoonida - a group of generalist 'ultimate opportunists', which infect many fished and aquacultured animals, as well as a broad suite of host taxa, including humans. In this study, we provide phylogenetic, ultrastructural, developmental, and pathological evidence for the creation of a new genus and species to hold a microsporidian parasite of the Jonah crab, Cancer borealis. Cancer borealis represents a species of commercial interest and has become the target of a recently developed fishery on the USA and Canadian Atlantic coast. This species was found to harbour a microsporidian parasite that develops in the cytoplasm of alpha and beta cells of the hepatopancreas. We retrieved a 937 bp fragment of the parasite SSU region, alongside developmental and ultrastructural data that suggests this species is âˆ¼ 87 % similar to Parahepatospora carcini and develops in a similar manner in direct association with the host cell cytoplasm. The mature spores are ovoid in shape and measure 1.48 ± 0.15 µm (SD) in length and 1.00 ± 0.11 µm (SD) in width. Phylogenetically, the new parasite clades in the Enterocytozoonida on the same branch as P. carcini. We provide a new genus and species to hold the parasite: Pseudohepatospora borealis n. gen. n. sp. (Microsporidia: Enterocytozoonida) and explore the likelihood that this species may fit into the Hepatoporidae family.

Host-parasite interactions: a study on the pathogenicity of different Mesanophrys sp. densities and hemocytes-mediated parasitic resistance of swimming crabs (Portunus trituberculatus).

Mesanophrys sp. is a parasitic ciliate that invades and destroys the hemocytes of the swimming crab (Portunus trituberculatus). In the present study, we employed an in vitro model to elucidate how Mesanophrys sp. destroys crab hemocytes. We also evaluated the relationship between the parasite's density, the destruction rate of the hemocytes, and the rapid proliferation pattern of parasites in host crabs. We found that the survival rate and cell integrity of crab hemocytes decreased with an increase in Mesanophrys sp. density, depicting a negative correlation between hemocyte viability and parasite density. Further analyses revealed that crab hemocytes could resist destruction by a low density (10 ind/mL) of Mesanophrys sp. for a long time (60 h). Mesanophrys sp. and its culture medium (containing the ciliate secretions) destroy the host hemocytes. The natural population growth rate of Mesanophrys sp. decreased with an increase in the parasite density, but the Mesanophrys sp. density did not affect the generation time of the parasites. In summary, Mesanophrys sp. can destroy crab hemocytes, and the degree of destruction is directly proportional to the parasite density. The resistance of crab hemocytes to Mesanophrys sp. decreased gradually with an increase in the parasite density.

Parasitic dinoflagellate Hematodinium in marine decapod crustaceans: a review on current knowledge and future perspectives.

Parasitic dinoflagellates of the genus Hematodinium are known to infect various marine crustaceans worldwide, especially crabs and several species of shrimp and lobster. Some of these species are new host species and components of commercial fishery products. These parasitic species are predominantly found in the hemolymph of the host and cause pathological changes and functional damage to organs and tissues, leading to death. In recent years, these parasites have infected important commercially valuable species, particularly in European waters, US waters, Australian waters, and recently in Shandong Peninsula in China. These Hematodinium pathogens were also reported to affect wild shrimp in Chinese waters and in the English North Sea. These rapid spreads affect crustacean aquaculture industries, where they are indeed a significant threat to the sustainability of the aquaculture of important crustaceans. The fishery products industries are also under pressure from the invasion of this pathogen, as the crab meat produced has a bitter taste, which may reduce its marketability. In response to these threats, this review was aimed at providing a broader understanding of the development of parasite distribution and ecological aspects of Hematodinium. In addition, the interaction of these pathogens with their hosts, the environmental drivers of Hematodinium disease, and future research perspectives were discussed.

Sacculina carcini impact on energy content of the shore crab Carcinus maenas L.

The impact of Sacculina carcini infection on the nutritional status of the shore crab Carcinus maenas was investigated in the western Dutch Wadden Sea for a period of 20 months. About 3.3% of the population was sacculinized, i.e. externally infected with S. carcini and only 0.7% presented scars of previous infection. The results of mixed linear models showed that sacculinized and non-sacculinized crabs had similar morphometric condition, while the energy density of parasitized crabs (externa excluded) was significantly reduced by about 4.3% overall, and by up to 5.8% in crabs under 40 mm carapace width. However, when Sacculina externa was included in the energy determinations, the difference in energy density decreased to 1.2%, while total energy content of the pair infected crab-parasite including externa was 30.8% higher than non-sacculinized crabs of similar size. The total energy content of ovigerous females (eggs included) was even higher, near doubling the energy of similar-sized crabs. The same way, total energy content of Sacculina externa was about 4 times lower than total energy of egg mass. The results suggest that the rhizocephalan parasite is efficient in consuming the energy that the host may allocate for growth and maintenance, but require future studies to disentangle the impact of the degree of internal infection and the implications for the dynamics of the population.

ß-1,3-Glucan from Euglena gracilis as an immunostimulant mediates the antiparasitic effect against Mesanophrys sp. on hemocytes in marine swimming crab (Portunus trituberculatus).

ß-1,3-glucans, natural polysaccharide groups, exert immunomodulatory effects to improve the innate response and disease resistance in aquatic species and mammals. However, this ß-glucan stimulant is yet to be assayed in swimming crab (Portunus trituberculatus) hemocytes. In this study, we explored the immunomodulatory effect of ß-1,3-glucans (derived from Euglena gracilis) via in vitro 24 h stimulation assays in swimming crab hemocytes. We found that this algal ß-1,3-glucans in crab hemocytes significantly elevated cellular enzymes related parameters, including phenoloxidase (PO), lysozyme, acid phosphatase (ACP) activities, and superoxide anion generation (O2-) rate both at intracellular (P < 0.05) and extracellular (P < 0.05) levels. Besides, alkaline phosphatase (AKP) in hemocytes exhibited no significant differences across the groups (P > 0.05). ß-glucan significantly influenced (P < 0.05) the activities of the antioxidant enzyme, superoxide dismutase (SOD) in hemocytes. Moreover, the relative mRNA expression of numerous immune-related genes, including proPO, TLR-2, Alf-1, NOX, Lysozyme, Crustin-1, and Cuznsod, was significantly higher stimulated hemocytes than in control (P < 0.05). We also reported the dose-dependent antiparasitic activity against Mesanophyrs sp., in stimulated hemocytes than in the control (P < 0.05). The present study collectively demonstrated that ß-glucan potentially stimulates innate immunity by elevating cellular enzyme responses and up-regulating the mRNA expression of genes associated with crab innate immunity. Thus, ß-glucan is a promising immunostimulant for swimming crab farming in crustaceans aquaculture.

Effect of environmental factors on survival and population growth of ciliated parasite, Mesanophrys sp. (Ciliophora: Scuticociliatia) infecting Portunus trituberculatus.

Mesanophrys sp. is a newly identified parasitic ciliate infecting farmed swimming crab. To demonstrate the correlation between parasite development and environmental conditions, this study aimed to investigate the effect of temperature, salinity, pH and frequency of passage of parasite on survival, growth and body size of Mesanophrys sp. in vitro. The results revealed that survival, population density and growth rate of the parasite were highest at 12°C and decreased with increasing temperature from 16 to 26°C. In addition, the survival, population density and growth rate of Mesanophrys sp. were high at 20‰. When salinity was adjusted to levels lower (0-10‰) and higher (40-60‰) than 20‰, the parasite's survival and growth rate gradually declined. The optimal pH for parasite survival was 8.0, whereas its survival was inhibited at <4.5 or >9.5. Our result also showed that parasite body proportions (length:width) were significantly smaller at the highest temperature compared to the lower temperature, whereas different salinities had no significant effect. Furthermore, we introduced dynamic parasite culture systems in vitro where Mesanophrys sp. was cultured in medium-containing culture plates through continually reducing and halving the old medium into fresh. Application of this optimized dilution timing technique with fresh medium and sub-cultured enabled a continuous culture of parasites. Under this optimized condition, the highest population density and exponential growth rate of the parasite were achieved than that of a control group. This study will help to understand the ciliated parasite infection dynamics and provides new possibilities for in vitro parasite-associated studies.

Parasitism-Induced Intersexuality in a Sexually Dimorphic Varunid Crab, Ptychognathus ishii (Decapoda: Varunidae).

Although many animals that perform sexual reproduction exhibit sexual dimorphism, individuals with intersex traits between the traits of males and females appear in some species, depending on environmental factors. Ptychognathus ishii, a varunid crab, exhibits distinctive sexual dimorphism in the morphology of its abdomen, chelipeds and setal tufts on the chelipeds. In this study, however, we report for the first time that intersex individuals with intermediate characters between those of males and females were occasionally found in wild populations. Morphological features of intersex individuals are described. Their taxonomic positions are identified based on DNA sequences of part of the mitochondrial cytochrome c oxidase I (COI) gene. It was shown that the intersexuality was induced by entoniscid parasites, because all intersex individuals were parasitized by entoniscid isopods, identified as Entionella sp. The apparent correlation between parasitism and morphological anomalies suggests that the parasitic isopods affect physiological conditions, leading to the feminization of male hosts.

Panopeispora mellora n. gen. n. sp. (microsporidia) infecting Say's crab (Dyspanopeus sayi) from the Atlantic shoreline of Canada.

Say's mud crab, Dyspanopeus sayi (Brachyura: Panopeidae) is a native shallow subtidal and inter-tidal inhabitant of the Atlantic coastline of North America and an invasive species in the Mediterranean and Black Seas. Little is known about the microparasites of this host and the broader Panopeidae. We describe a novel microsporidian parasite infecting the musculature of D. sayi from Malagash, Nova Scotia (Canada), at a prevalence of 7%. Histopathology and molecular diagnostics were used to describe pathology and parasite phylogenetics, respectively. Based on SSU rDNA gene sequencing we propose that the microsporidian requires establishment of a new genus (Panopeispora n. gen.) and species (Panopeispora mellora n. sp.), due to significant differences to closest known taxa (e.g. Facilispora margolisi [81% similarity] and Thelohania butleri [80% similarity]), residing in Clade V of the Microsporidia. Archived, wax-embedded histological material was re-processed for transmission electron microscopy to obtain preliminary details of its intracellular development cycle and ultrastructure within the host musculature. The discovery of this pathogen is discussed with relevance to microsporidian taxonomy and the potential for achieving ultrastructural data from archived material.

First detection of Hematodinium sp. In spiny king crab Paralithodes brevipes, and new geographic areas for the parasite in tanner crab Chionoecetes bairdi, and red king crab Paralithodes camtschaticus.

A parasitic dinoflagellate of the genus Hematodinium was found off the Pacific coast of Kamchatka in three species of crabs: red king crab Paralithodes camtschaticus, tanner crab Chionoecetes bairdi, and spiny king crab Paralithodes brevipes. This is the first detection of Hematodinium in spiny king crab. The results of the genetic analysis showed that the pathogen found in P. brevipes, P. camtschaticus, and C bairdi from the Avacha and Kronotsky bays off the Pacific coast of Kamchatka was the same or very close to the Hematodinium sp., which infects many species of crustaceans in the Northern Hemisphere. The prevalence of infection was 0.2% for tanner crabs and 2.7% for red king crabs. Due to a limited sample size, we were unable to calculate the prevalence for spiny king crabs and female red king crabs. Both the macroscopic and microscopic signs of the pathology were similar in all diseased crabs. The differences in the micromorphology of the Hematodinium cells we found in the three crab species, including the presence or absence of trichocysts, the shape of the plasmodia, and the structure of pore complexes, are most likely related to the life cycle and the physiology of the parasite. The results of the genetic analysis showed that the pathogen found in P. brevipes, P. camtschaticus, and C. bairdi from the Avacha and Kronotsky bays of the Pacific coast of Kamchatka was the same or very close to the Hematodinium sp., which infects many species of crustaceans in the Northern Hemisphere.

Symbionts of invasive and native crabs, in Argentina: The most recently invaded area on the Southwestern Atlantic coastline.

Biological invasions have the capacity to introduce non-native parasites. This study aimed to determine whether the invasive green crab population, Carcinus spp., on the Southwestern Atlantic coast of Argentina harbours any symbionts, and whether these may spillover or spillback between native crabs, Cyrtograpsus altimanus and C. angulatus. Macroscopy, histology, and molecular analyses of some parasites were used to describe and compare their diversity across the three species of crab. We also evaluated the susceptibility of invasive Carcinus spp. to a native digenean, Maritrema madrynense, via experimental infections (exposure and cohabitation). Our results revealed that the green crab pathobiome included similar symbiotic groups to native crabs. This included putative viral, bacterial, and protozoan parasites. Haplosporidium-like observations were recorded in all crab species, and a single green crab was found to be parasitized by an Agmasoma-like microsporidium. Metagenomic analysis of one individual revealed additional symbiotic diversity (46 bacteria, 5 eukaryotic species). The green crabs were infected by more microparasite taxa than the native crabs (5:3). Wild populations of Carcinus spp. were free of metazoan parasites and are shown not to be susceptible to M. madryense under experimental conditions. Our results suggest a reduction/escape of macroparasites (trematode Maritrema madrynense; acanthocephalan Profilicollis chasmagnathi) in invasive Carcinus spp. compared to their native competitors.

The parasitic dinoflagellate Hematodinium infects multiple crustaceans in the polyculture systems of Shandong Province, China.

The parasitic dinoflagellates of the Hematodinium genus have impacted wild and cultured stocks of commercial crustaceans worldwide. In the past decade, outbreaks of Hematodinium epizootics resulted in substantial mortalities in cultured Chinese swimming crabs Portunus trituberculatus in the polyculture ponds located in Shandong Peninsula, whereas the source and transmission of the parasite in the polyculture pond system remains to be determined. During April to December of 2018, 2034 crabs and 108 shrimps were collected from the polyculture pond systems in the highly endemic area of Hematodinium diseases in Qingdao, Shandong Province. Among those, 188 individuals of the 6 crab species were infected by the parasite, including 4 novel host species (Uca arcuate, Hemigrapsus penicillatus, Helice wuana and Macrophthalmus japonicas). No infection was identified in Penaeus monodon. Further phylogenetic analyses indicated that the Hematodinium isolate infecting the six crab hosts, together with other isolates reported from China, composed the genotype II of Hematodinium perezi. The parasite was more infectious to cultured Portunus trituberculatus and the dominant wild crab Helice tientsinensis dwelling in the waterways connecting to the polyculture ponds, even though it was found to be a host generalist pathogen. The prevalence of Hematodinium perezi infection in Helice tientsinensis was higher than that of other wild crabs and showed significant positive correlation with that of the cultured Portunus trituberculatus. The results indicated that the wild crabs, particularly Helice tientsinensis, were the important alternate hosts closely involved in transmission and spreading of the Hematodinium disease in the polyculture pond systems.

Two new species of Unikaryon (Microsporidia) hyperparasitic in microphallid metacercariae (Digenea) from Florida intertidal crabs.

The genus Unikaryon (Microsporidia) holds exclusively hyperparasites of Platyhelminthes. Four species of Unikaryon are presently known from trematodes infecting mollusks and fish, and one from a cestode infecting a fish. Here we report two species of Unikaryon from microphallid trematode metacercariae parasitizing the brachyuran crabs, Panopeus herbstii and Pachygrapsus transversus, collected from intertidal habitats in Florida. The first microsporidium, which we assign here to a new species, Unikaryon panopei sp. n., was isolated from Microphallus sp. encysted in Panopeus herbstii from Tampa Bay. The specific designation for the second Unikaryon sp. (Unikaryon sp. 2), which occurred in metacercaria of Diacetabulum sp. found in P. transversus from the Florida Keys, is pending due to the lack of SSrDNA sequence data. Light and electron microscopy demonstrates that both species display characteristics of the genus Unikaryon including the arrangement of spores in sets of two, large posterior vacuole, and eccentric position of the polar filament. Spores of Unikaryon panopei sp. n., unlike those of Unikaryon sp. 2, assemble in large membrane-bound masses containing hundreds of organisms, and display a larger number of polar filament coils - 7-8, compared to 4-5 in Unikaryon sp. 2 The SSUrDNA-inferred phylogenetic analysis places Unikaryon panopei in one clade with Unikaryon legeri, the only other molecularly characterized member of the genus, with 94% of SSUrDNA similarity. These findings increase the number of species parasitizing trematodes and broaden the host range of Unikaryon spp.

Dynamic transcriptome landscape of Paragonimus proliferus developmental stages in the rat lungs.

Paragonimus proliferus, a lung fluke of the genus Paragonimus, was first reported in Yunnan province, China. P. proliferus can infect Sprague-Dawley (SD) rats and cause lung damage, but there is still no direct evidence of human infection. Until now, there has been a lack of studies on P. proliferus parasitism and development in mammalian lung tissue. The aim of this study was to perform transcriptomic profiling of P. proliferus at different developmental stages. SD rats were infected with P. proliferus metacercariae obtained from crabs; worms isolated from the lungs at different time points as well as metacercariae were subjected to whole transcriptome sequencing. Overall, 34,403 transcripts with the total length of 33,223,828 bp, average length of 965 bp, and N50 of 1833 bp were assembled. Comparative analysis indicated that P. proliferus, similar to other Paragonimus spp., expressed genes related to catabolism, whereas P. proliferus-specific transcripts were related to the maintenance of cellular redox homeostasis, sensitivity to bacteria, and immune response. Transcriptional dynamics analysis revealed that genes involved in the regulation of catabolism and apoptosis had stable expression over the P. proliferus life cycle, whereas those involved in development and immune response showed time-dependent changes. High expression of genes associated with immune response corresponded to that of genes regulating the sensitivity to bacteria and immune protection. We constructed a P. proliferus developmental model, including the development of the body, suckers, blood cells, reproductive and tracheal systems, lymph, skin, cartilage, and other tissues and organs, and an immune response model, which mainly involved T cells and macrophages. Our study provides a foundation for further research into the molecular biology and infection mechanism of P. proliferus.