Free-living Stages Research Articles

Search, find, and penetrate: ultrastructural data of furcocercariae of Cardiocephaloides longicollis (Digenea, Strigeidae) explain their transmission and infection strategy into fish hosts.

The present study provides an overview of the structures linked to fish host finding, recognition, and invasion of one of the most commonly occurring morphotypes among trematodes, furcocercariae. For this, we use free-swimming cercariae of the strigeid Cardiocephaloides longicollis (Rudolphi 1819) Dubois, 1982. Their elongated cercarial body and bifurcated tail are covered by a tegument with an irregular surface, showing numerous folds arranged in different directions and a typical syncytial organization. Both the body and the bifurcated tail are covered with short spines, rose-thorn shaped, as well as four types of sensory papillae, distinguished by the presence or absence of a cilium, its length, and their position on the cercarial body. These papillae are especially important for free-living stages that rely on external stimuli to locate and adhere to the host. A specialized anterior organ is located at the anterior part of the cercariae and is encircled by a triangle-shaped group of enlarged pre-oral spines followed by a transverse row of enlarged post-oral spines that, together with the sensory papillae, allow active finding, recognition, and penetration into fish. The ventral sucker, covered with inner-oriented spines, sensory papillae, and cilia, helps during this process. The cercariae of C. longicollis possess three types of gland cells (a head gland and two types of penetration glands), each containing different types of secretory granules that play a role in host invasion. The protonephridial excretory system consists of an excretory bladder, a system of collecting tubules, flame cells, and two excretory pores in the middle of each furcae, which serve to control osmoregulation in their marine environment, as well as to eliminate metabolic waste. Together with the four types of sensory endings, the central ganglion forms the nervous system. Our results add novel information on the ultrastructure of strigeid furcocercariae, being essential to interpret these data in relation of their functional role to better understand the transmission and penetration strategies that cercariae display to infect their fish hosts.

Pasture rewetting in the context of nature conservation shows no long-term impact on endoparasite infections in sheep and cattle

BackgroundNature conservation with reduced drainage of pastures has been increasingly promoted in agriculture in recent years. However, moisture on pastures is a crucial factor for the development of free-living stages of many parasite species in ruminants. Hence, for the first time, we conducted a field study between 2015 and 2017 at the German North Sea coast to investigate the long-term effect of pasture rewetting (since 2004) on endoparasite infections in sheep and cattle.MethodsWe examined faecal samples of 474 sheep and 646 cattle from five farms in spring, summer and autumn each year for the presence of endoparasite infections. Animals were kept on conventionally drained, undrained and rewetted pastures. The association between pasture rewetting and endoparasite infection probability was analysed in generalized linear mixed models and including further potential confounders.ResultsInfection frequencies for gastrointestinal strongyles, Eimeria spp. and Strongyloides papillosus were significantly higher in sheep (62.9%, 31.7% and 16.7%) than in cattle (39.0%, 19.7% and 2.6%). Fasciola hepatica was detected with a frequency of 13.3% in sheep and 9.8% in cattle, while rumen fluke frequency was significantly higher in cattle (12.7%) than in sheep (3.8%). Nematodirus spp., lungworms (protostrongylids, Dictyocaulus viviparus), Moniezia spp., Trichuris spp. and Dicrocoelium dendriticum were identified in less than 7% of samples. Co-infection with more than three endoparasite taxa was present significantly more often in sheep than in cattle. We identified significant positive correlations above 0.2 for excretion intensities between S. papillosus with strongyles, Eimeria spp. and Nematodirus spp. in sheep and between strongyles and Nematodirus spp. in cattle. Pasture rewetting had no long-term effect on endoparasite infections, neither in sheep nor in cattle. Interestingly, F. hepatica infections decreased significantly in sheep and cattle from 2015 (10.9% and 13.9%) to 2017 (1.4% and 2.1%).ConclusionsPasture rewetting for nature conservation did not increase endoparasite infection probability in ruminants in the long term. This finding should be confirmed in ongoing studies aimed at further animal welfare parameters. The rapid decrease in F. hepatica infections over 3 years may suggest climatic impact or competition with rumen flukes in addition to potential anthelmintic treatment after feedback of the results to the farmers.Graphical

The enemy of my enemy is my friend: Consumption of parasite infectious stages benefits hosts and predators depending on transmission mode.

Research Highlight: Hobart, B. K., Moss, W. E., McDevitt-Galles, T., Stewart Merrill, T. E., Johnson, P. T. J. (2021). It's a worm-eat-worm world: Consumption of parasite free-living stages protects hosts and benefits predators. Journal of Animal Ecology, https://doi.org/10.1111/1365-2656.13591 Many parasites and pathogens have infectious stages that are vulnerable to consumption by non-host organisms. This consumption can benefit both the predators that consume this unusual food resource and the hosts which are less likely to encounter infectious propagules. Yet the importance of these benefits may also depend on parasite transmission, which can influence consumer opportunities to feed upon free-living infectious stages. Hobart et al. (2021) report that freshwater snails with high densities of symbiotic oligochaetes are less likely to be parasitized by trematodes (Platyhelminthes) with an 'active' versus 'passive' (motile or stationary, respectively) mode of transmission, supporting a protective effect via oligochaete predation upon infectious propagules. However, these predators benefit from snails harbouring passively acquired infections, and likely achieve their higher abundance from easy access to prey in the form of a second infectious stage emerging from these hosts. Consumption of free-living infectious stages is thus beneficial to hosts and predators, but varies with parasite life history and ecology.

Models of reproduction, development, and growth in invertebrates

Growing and producing offspring are inherent to all living organisms. The life cycle of an invertebrate species typically consists of four distinct stages: egg, larva, juvenile, and adult. In spite of this, there can be variations in this life cycle. Marine invertebrates are composed of microscopic, free-living dispersive stages, which may feed (planktotrophic) or not feed (lecithotrophic). This paper outlines a simple model to explain how reproductive patterns have evolved in marine benthic invertebrates.

Risk of Human Exposure to the Intestinal Schistosome, Schistosoma mansoni, across Seasons along the Senegal River

Background: Schistosomiasis is an emerging disease associated with changes to the environment that have increased human contact rates with disease-causing parasites, flatworms that are released from freshwater snails. For example, schistosomiasis remains a major public health problem in Northern Senegal, where prevalence in schoolchildren often reaches 90%. Aim: This study focuses on the impact of seasonality on the risk of human exposure (RHE) to Schistosoma mansoni, defined as the total number of cercariae (the free-living life stage that infects humans) shed from all Biomphalaria pfeifferi snails collected at a site using standardized methods. We focus on RHE because it is rarely quantified and a recent study demonstrated that snails stop shedding cercariae when snail densities increase and thus per capita snail resources become limited [2], suggesting that densities of snails might not be directly proportional to RHE to schistosomes. Method: We sampled four water access points in three villages every other week during the early (Dry1) and later dry seasons (Dry2) and the rainy season, quantifying the abundance of infected and non-infected snail intermediate hosts, cercariae released per infected snail, and water chemistry. We used simple and multiple linear regressions to assess how seasonality and environmental parameters affect non-infected and infected snail abundance and RHE. Results: Although RHE was found across all seasons, the abundance of infected and non-infected snail intermediate hosts and cercariae, as well as prevalence (23.71%), were all highest in the rainy season. In the rainy season, RHE was positively associated with the density of snail hosts and their periphyton food resource. Conclusion: Although previous studies have examined the influence of seasonality on snail densities, few studies have explored the effects of seasonality on cercarial densities, which is the primary source of infection to humans. Our study demonstrates that cercarial densities are greater in the rainy season than in the early or late dry seasons. Given that cercarial densities directly pose risk of infection to humans, unlike non-infected or infected snails, these finding should help to inform decision making and schistosomiasis control efforts in West Africa.

Assessment of ecological conditions for growing the market-size fish in the Kantivka fattening pond (Khmelnychchyna, Ukraine)

Abiotic (water quality) and biotic (Phyto-, zooplankton and benthos) factors in the Kantivka fattening pond the Khmelnitskrybhosp Private Joint-Stock Company were studied during the vegetation seasons in 2020. The aquaculture objects in the pond are cyprinid fish species (carp, grass carp, silver carp, pike). The ecological conditions of the pond were evaluated as good enough for the fish fattening purpose. The critical water quality parameters were following traditional values accepted for fish farming in Ukraine. The taxonomic composition of plankton and benthos was represented by widespread and Ubiquiti species mainly. The 124 species and intraspecies taxa of microalgae and the 34 species of zooplankton were identified in the Kantivka fattening pond. The mean abundance and biomass for the phytoplankton were 183.8 million cells/dm3 and 35.61 mg/dm3, respectively; for zooplankton, those were as 336600 ind/m3 and 14.83 g/m3, and for zoobenthos, those values were as 309.6 ind/m2, and biomass as 5.84 g/m2. Such parameters of natural forage base correspond to the optimal values for natural fish food according to fish farming standards in Ukraine. In the late summer, the free-living copepod stages and mature adult females of the parasitic copepod crustacean Ergasilus sieboldi Nordmann were registered in plankton. This copepod causes the invasion of fish diseases like infection and destruction of gill arches, which leads to a severe disturbance in fish respiration, emaciation, delaying in growth, and the potential death of commercial fish even. The development of natural fish food in the Kantivka fattening pond is shown an intensive level of biomass and abundance of phytoplankton, zooplankton, and zoobenthos. It is considered sufficient to meet the natural nutritional needs (in combination with artificial feeding) for older age groups of the cultivated cyprinid fish species.

First report of the order Mysida (Crustacea) in Antarctic marine ice caves, with description of a new species of Pseudomma and investigations on the taxonomy, morphology and life habits of Mysidetes species.

SCUBA diving explorations of three islands off Dumont d’Urville Station at the coast of Adélie Land, East Antarctica, enabled the observation of marine ice caves. Sampling in this unusual habitat yielded a total of three species of Mysidae, altogether previously poorly known or unknown to science. Pseudommakryotroglodytumsp. nov. is described, based on the structure of the antennal scale, telson and on cornea-like lateral portions set off against the main body of eyeplates. Mysidetesilligi is re-established at species level after almost a century in synonymy. Re-descriptions are provided for M.illigi and M.hanseni, based on types and ice cave materials. Keys to the Southern Ocean species of Pseudomma and to the world-wide species of Mysidetes are given.Phylogenetic trees are provided for the genera Pseudomma and Mysidetes. 18S rDNA sequences of P.kryotroglodytum differ from GenBank sequences of other Pseudomma species. First sequence data are given for species of the genus Mysidetes: 18S differs between the two examined species and COI is quite diverse between and within species.We found previously unknown, probably sensorial structures in these ice cave species: in P.kryotroglodytum, the basal segment of the antennula shows a pit-like depression with striated pad on the bottom and a median cyst, connected with the bottom of the eyeplate cleft. M.illigi shows a female homologue of the appendix masculina bearing a field of modified setae. Subsequent investigations demonstrated these structures also in species from other habitats.The feeding apparatus and stomach contents of the three ice cave species point to brushing of small particles (detritus, microalgae) from available surfaces, such as sediment, rock and the ice surface. Differences in the feeding apparatus are very subtle between the two Mysidetes species. The high content of fat bodies in M.hanseni could help it to survive periods of starvation. The large storage volume of the foregut in P.kryotroglodytum points to the collection of food with low nutritional quality and could help to balance strongly fluctuating food availability.Summer specimens of M.hanseni showed a bimodal frequency of developmental stages in the marsupium and bimodal size-frequency distribution of free-living stages. The females with younger brood (embryos) were, on average, larger and carried more marsupial young than those with older brood (nauplioid larvae). All examined incubating and spent females showed (almost) empty foreguts and empty ovarian tubes, suggesting possible semelparity and death following the release of young. The absence of juveniles and immature females from summer samples suggests that growth and accumulation of fat and yolk occur outside ice caves, while such caves could be used by fattened adults as shelter for brooding. A provisional interpretation proposes a biannual life cycle for M.hanseni, superimposed with shifted breeding schedules, the latter characterised by early breeding and late breeding females, probably in response to harsh physical and trophic conditions along the continental coast of Antarctica.

Features of bacterial diseases of rainbow trout in industrial farms in North-West Russia

Studies of the parasite fauna of rainbow trout have revealed a number of pathogens of parasitic diseases at enterprises that use open water bodies as sources of water supply. Of the parasitic protozoa, the most dangerous is Ichthyophthirius multifiliis. This is due to the presence of a free-living stage in this ciliate, as well as to the possibility of localization under the fish scales. Of the helminthiases that occur in rainbow trout during artificial cultivation on the farms of the North-West region of Russia, the most problematic are monogenes and cestodes, which cause gyrodactylosis and trienophorosis. The plerocercoids of the cestode Triaenophorus crassus, in particular, are localized in the musculature of fish and, with a high degree of invasion, trout loses its commercial value. In the presence of a focus of parasitic diseases in natural reservoirs, wild fish are a reservoir of invasion, aquatic invertebrates (planktonic crustaceans, molluscs) are the most important element of the parasite-host system, and fish-eating birds, being the final host, contribute to the spread of invasions. In cage farms, due to the peculiarities of the cultivation technology, the fight against parasitic protozoa is extremely difficult and requires significant labor costs. In basin farms, on the contrary, the design of fish-breeding tanks and the water supply system allows for the most efficient implementation of therapeutic and prophylactic measures. Nevertheless, outbreaks of parasitic diseases can be observed in violation of the cultivation technology (overcompaction of the planting, violation of the feeding regime, etc.). Control of the epizootic state makes it possible to identify infection and carry out a set of necessary therapeutic and preventive measures.

Biological Invasion of Fish Parasite Neoergasilus japonicus (Harada, 1930) (Copepoda: Ergasilidae) in Lake Grand Laoucien, France: A Field Study on Life Cycle Parameters and Reasons for Unusual High Population Density.

The fish parasite, Neoergasilus japonicus (Harada, 1930), native to Eastern Siberia and the Amur River catchment area, invaded European water bodies in the middle of the last century, possibly due to the human-mediated distribution of fish in the Amur complex (i.e., the genera Hypophthalmichthys and Ctenopharyngodon). In the deep karst lake, Grand Laoucien (Marseille area, France), this species had an unusually high population density (from 1000 ind./ m3 in zooplankton to 4000 ind./ m3 in the nearshore area) during the free-living period of its life cycle. The annual cycle of N. japonicus includes a 5-month overwintering of fertilized females attached to fish fins and, following this, a five- to six-generation chain from March to November, when the free-living stages in the population alternate with parasite females which attach to their hosts for breeding. The population density of the parasites in zooplankton increased exponentially from spring to autumn, which positively correlated with temperature. We found a strong correlation between N. japonicus density and the community development of microphytobenthos, but not between N. japonicus and phyto- or zooplankton dynamics. The local contributing factors included a seasonal three-fold decrease in water levels and the development of anoxia in profundal waters, which led to a high ambient fish density and thus susceptibility to the parasite. Although the free-living parasite represented only 1% of zooplankton production, it consumed up to 25% of small invertebrate productivity. The maximum intensity of infection reached 140 parasites per fish, or 4.14 per g of weight. The high infection of fish with this parasite, in our opinion, indicated the danger it poses to the local ichthyofauna, which first encountered this new parasite.

Review: Schistosoma mansoni phosphatidylinositol 3 kinase (PI3K)/Akt/mechanistic target of rapamycin (mTOR) signaling pathway.

Schistosoma mansoni worms are under a milieu of external and internal signaling pathways. The life-cycle stages are exposed to enormous stimuli within the mammalian and the snail hosts and as free-living stages in the fresh water. Furthermore, there is a unique interplay between the male and the female worms involving many stimuli from the male essential for full development of the female. PI3K/Akt/mTOR is an evolutionarily divergent signal transduction pathway universal to nearly every multicellular organism. This work reviews the Schistosoma mansoni PI3K/Akt/mTOR signal pathways and the involvement of the signal in the worms' physiology concerning the uptake of glucose, reproduction and survival. The inhibitors of the signal pathway used against Schistosoma mansoni were summarized. Given the importance of the PI3K/Akt/mTOR signal pathway, its inhibition could be a promising control strategy against schistosomiasis.

Comparative phylogeography of two bat species and their mites in Iran shows impact of host sociality and vagility on population structure

Parasite species lacking a free-living stage rely on their hosts for dispersal. Their population genetic structure depends on the host's vagility and dispersal rate. To gain more insight into the drivers responsible for shaping the spatio-temporal population structure in host–parasite systems, we used mitochondrial DNA sequences to compare patterns of genetic diversity in two closely related and contact-transmitted parasitic wing mites Spinturnix psi and S. myoti with their bat hosts Miniopterus pallidus and Myotis blythii, respectively, across vast distances in Iran. We observed almost no genetic differentiation between mites living on bats in different colonies even from distant locations, whereas we found some level of genetic differentiation and isolation by distance in each host species, particularly in the less vagrant M. blythii. Despite Iran's high spatial divergence and long distance between the sampled locations, local genetic diversity and inter-population gene flow in the parasites were high, even between different sides of the Zagros and Alborz Mountains. The genetic similarity that was observed among mite populations likely reflects genetic exchanges between colonies at swarming places of bats, as well as the possible occupation of other host species, resulting in a higher effective population size and more dispersal opportunities for the mites.

High-throughput sequencing of Fasciola spp. shows co-infection and intermediate forms in Balochistan, but only Fasciola gigantica in the Punjab province of Pakistan.

Fasciola gigantica and Fasciola hepatica are digenetic trematodes causing fasciolosis in ruminants. The host and geographical distribution of both Fasciola species are influenced by environmental and climatic conditions favouring survival and development of free-living stages and intermediate hosts, and livestock management practices. The aim of the present study was to describe the host distribution of the two Fasciola species in buffalo, cattle, goats, and sheep in the Balochistan and Punjab provinces of Pakistan. 359 flukes were collected from a total of 32 livers from the four livestock species. Deep amplicon sequencing of the internal transcribed spacer region 2 of ribosomal DNA (rDNA ITS-2) and mitochondrial nicotinamide adenine dinucleotide dehydrogenase 1 (mtDNA ND-1) loci confirmed co-infection of F. hepatica and F. gigantica in Balochistan and single species F. gigantica infection in Punjab. In Balochistan, co-infections and hybrids of both Fasciola species were identified in cattle, with more F. hepatica detected than F. gigantica. However, F. hepatica was the only species identified in goats, and F. gigantica was the only species identified in buffalo. In Punjab, all flukes were confirmed as F. gigantica in each of the four livestock species. Overall, the results indicate differences in the host and geographical distribution of F. gigantica and F. hepatica, and provide useful knowledge for the development of control strategies for livestock and humans.

It's a worm-eat-worm world: Consumption of parasite free-living stages protects hosts and benefits predators.

Predation on parasites is a common interaction with multiple, concurrent outcomes. Free-living stages of parasites can comprise a large portion of some predators' diets and may be important resources for population growth. Predation can also reduce the density of infectious agents in an ecosystem, with resultant decreases in infection rates. While predator-parasite interactions likely vary with parasite transmission strategy, few studies have examined how variation in transmission mode influences contact rates with predators and the associated changes in consumption risk. To understand how transmission mode mediates predator-parasite interactions, we examined associations between an oligochaete predator Chaetogaster limnaei that lives commensally on freshwater snails and nine trematode taxa that infect snails. Chaetogaster is hypothesized to consume active (i.e. mobile), free-living stages of trematodes that infect snails (miracidia), but not the passive infectious stages (eggs); it could thus differentially affect transmission and infection prevalence of parasites, including those with medical or veterinary importance. Alternatively, when infection does occur, Chaetogaster can consume and respond numerically to free-living trematode stages released from infected snails (cercariae). These two processes lead to contrasting predictions about whether Chaetogaster and trematode infection of snails correlate negatively ('protective predation') or positively ('predator augmentation'). Here, we tested how parasite transmission mode affected Chaetogaster-trematode relationships using data from 20,759 snails collected across 4years from natural ponds in California. Based on generalized linear mixed modelling, snails with more Chaetogaster were less likely to be infected by trematodes that rely on active transmission. Conversely, infections by trematodes with passive infectious stages were positively associated with per-snail Chaetogaster abundance. Our results suggest that trematode transmission mode mediates the net outcome of predation on parasites. For trematodes with active infectious stages, predatory Chaetogaster limited the risk of snail infection and its subsequent pathology (i.e. castration). For taxa with passive infectious stages, no such protective effect was observed. Rather, infected snails were associated with higher Chaetogaster abundance, likely owing to the resource subsidy provided by cercariae. These findings highlight the ecological and epidemiological importance of predation on free-living stages while underscoring the influence of parasite life history in shaping such interactions.

Characterizing parasitic nematode faunas in faeces and soil using DNA metabarcoding

BackgroundGastrointestinal parasitic nematodes can impact fecundity, development, behaviour, and survival in wild vertebrate populations. Conventional monitoring of gastrointestinal parasitic nematodes in wild populations involves morphological identification of eggs, larvae, and adults from faeces or intestinal samples. Adult worms are typically required for species-level identification, meaning intestinal material from dead animals is needed to characterize the nematode community with high taxonomic resolution. DNA metabarcoding of environmental samples is increasingly used for time- and cost-effective, high-throughput biodiversity monitoring of small-bodied organisms, including parasite communities. Here, we evaluate the potential of DNA metabarcoding of faeces and soil samples for non-invasive monitoring of gastrointestinal parasitic nematode communities in a wild ruminant population.MethodsFaeces and intestines were collected from a population of wild reindeer, and soil was collected both from areas showing signs of animal congregation, as well as areas with no signs of animal activity. Gastrointestinal parasitic nematode faunas were characterized using traditional morphological methods that involve flotation and sedimentation steps to concentrate nematode biomass, as well as using DNA metabarcoding. DNA metabarcoding was conducted on bulk samples, in addition to samples having undergone sedimentation and flotation treatments.ResultsDNA metabarcoding and morphological approaches were largely congruent, recovering similar nematode faunas from all samples. However, metabarcoding provided higher-resolution taxonomic data than morphological identification in both faeces and soil samples. Although concentration of nematode biomass by sedimentation or flotation prior to DNA metabarcoding reduced non-target amplification and increased the diversity of sequence variants recovered from each sample, the pretreatments did not improve species detection rates in soil and faeces samples.ConclusionsDNA metabarcoding of bulk faeces samples is a non-invasive, time- and cost-effective method for assessing parasitic nematode populations that provides data with comparable taxonomic resolution to morphological methods that depend on parasitological investigations of dead animals. The successful detection of parasitic gastrointestinal nematodes from soils demonstrates the utility of this approach for mapping distribution and occurrences of the free-living stages of gastrointestinal parasitic nematodes.Graphical abstract

Phylogenomics of a new fungal phylum reveals multiple waves of reductive evolution across Holomycota

Compared to multicellular fungi and unicellular yeasts, unicellular fungi with free-living flagellated stages (zoospores) remain poorly known and their phylogenetic position is often unresolved. Recently, rRNA gene phylogenetic analyses of two atypical parasitic fungi with amoeboid zoospores and long kinetosomes, the sanchytrids Amoeboradix gromovi and Sanchytrium tribonematis, showed that they formed a monophyletic group without close affinity with known fungal clades. Here, we sequence single-cell genomes for both species to assess their phylogenetic position and evolution. Phylogenomic analyses using different protein datasets and a comprehensive taxon sampling result in an almost fully-resolved fungal tree, with Chytridiomycota as sister to all other fungi, and sanchytrids forming a well-supported, fast-evolving clade sister to Blastocladiomycota. Comparative genomic analyses across fungi and their allies (Holomycota) reveal an atypically reduced metabolic repertoire for sanchytrids. We infer three main independent flagellum losses from the distribution of over 60 flagellum-specific proteins across Holomycota. Based on sanchytrids’ phylogenetic position and unique traits, we propose the designation of a novel phylum, Sanchytriomycota. In addition, our results indicate that most of the hyphal morphogenesis gene repertoire of multicellular fungi had already evolved in early holomycotan lineages.

Cytochrome P450s Are Essential for Insecticide Tolerance in the Endoparasitoid Wasp Meteorus pulchricornis (Hymenoptera: Braconidae).

Simple SummaryP450s are involved in the detoxification process in insects; however, less attention has been paid for their roles in parasitoid wasps. Here, we identified 28 P450s from the previous constructed transcriptome dataset of Meteorus pulchricornis and validated their expression patterns under insecticides exposure. Furthermore, we silenced CYP369B3 in adult wasps and demonstrated that the knocking down CYP369B3 significantly increased of the mortality of adult wasps when they were exposed to each chemical insecticide, phoxim, cypermethrin, and chlorfenapyr, respectively. The present study provides a theoretical base for functional research on P450 genes in parasitoid wasps.With the widespread application of insecticides, parasitoid wasps may also be under risk when exposed to insecticides directly at their free-living stages. The endoparasitoid wasp Meteorus pulchricornis is the predominant natural enemy of many lepidopteran pests, such as Spodoptera litura and Helicoverpa armigera. The cytochrome P450 monooxygenases constitute a ubiquitous and complex superfamily of hydrophobic, haem-containing enzymes. P450s are involved in the detoxification of many xenobiotics. However, their exact roles in the tolerance mechanism in parasitoids toward insecticides has received less attention. Here, 28 P450 genes in M. pulchricornis were identified from a previously constructed transcriptome dataset. These P450 genes belonged to CYP2, -3, and -4, and mitochondrial clans. Subsequently, eight candidate MpulCYPs were selected from four CYP clans to validate their expression patterns under phoxim, cypermethrin, and chlorfenapyr exposure by qRT-PCR. The results showed that all three insecticides had significant effects on the expression of MpulCYPs. To further study the function of P450s, CYP369B3 was silenced, and its expression levels of CYP369B3 were significantly decreased. Survival analysis indicated that after dsRNA injection, the mortality rate of wasps was significantly increased when M. pulchricornis females were exposed to insecticides compared to control groups. Our findings provide a theoretical base for elucidating the mechanism of insecticide tolerance and promote functional research on P450 genes in parasitoid wasps.

Dinophyceae can use exudates as weapons against the parasite Amoebophrya sp. (Syndiniales)

Parasites in the genus Amoebophrya sp. infest dinoflagellate hosts in marine ecosystems and can be determining factors in the demise of blooms, including toxic red tides. These parasitic protists, however, rarely cause the total collapse of Dinophyceae blooms. Experimental addition of parasite-resistant Dinophyceae (Alexandrium minutum or Scrippsiella donghaienis) or exudates into a well-established host-parasite coculture (Scrippsiella acuminata-Amoebophrya sp.) mitigated parasite success and increased the survival of the sensitive host. This effect was mediated by waterborne molecules without the need for a physical contact. The strength of the parasite defenses varied between dinoflagellate species, and strains of A. minutum and was enhanced with increasing resistant host cell concentrations. The addition of resistant strains or exudates never prevented the parasite transmission entirely. Survival time of Amoebophrya sp. free-living stages (dinospores) decreased in presence of A. minutum but not of S. donghaienis. Parasite progeny drastically decreased with both species. Integrity of the dinospore membrane was altered by A. minutum, providing a first indication on the mode of action of anti-parasitic molecules. These results demonstrate that extracellular defenses can be an effective strategy against parasites that protects not only the resistant cells producing them, but also the surrounding community.

Exposure to dexamethasone modifies transcriptomic responses of free-living stages of Strongyloides stercoralis.

Hyperinfection and disseminated infection by the parasitic nematode Strongyloides stercoralis can be induced by iatrogenic administration of steroids and immunosuppression and lead to an elevated risk of mortality. Responses of free-living stages of S. stercoralis to the therapeutic corticosteroid dexamethasone (DXM) were investigated using RNA-seq transcriptomes of DXM-treated female and male worms. A total of 17,950 genes representing the transcriptome of these free-living adult stages were obtained, among which 199 and 263 were differentially expressed between DXM-treated females and DXM-treated males, respectively, compared with controls. According to Gene Ontology analysis, differentially expressed genes from DXM-treated females participate in developmental process, multicellular organismal process, cell differentiation, carbohydrate metabolic process and embryonic morphogenesis. Others are involved in signaling and signal transduction, including cAMP, cGMP-dependent protein kinase pathway, endocrine system, and thyroid hormone pathway, as based on Kyoto Encyclopedia of Genes and Genomes analysis. The novel findings warrant deeper investigation of the influence of DXM on growth and other pathways in this neglected tropical disease pathogen, particularly in a setting of autoimmune and/or allergic disease, which may require the clinical use of steroid-like hormones during latent or covert strongyloidiasis.

Direct release of embryonic sporophytes from adult Nereocystis luetkeana (Laminariales, Ochrophyta) in a high latitude estuary

Kelp life history pathways alternate between macroscopic sporophytes that produce spores and microscopic gametophytes that produce gametes. Occasionally, an alternative pathway is seen. This study examined the circumstances by which the high latitude estuarine bull kelp, Nereocystis luetkeana, foregoes the “free-living microscopic stages by releasing embryonic sporophytes directly from sori. Sori were collected from adult N. luetkeana sporophytes from eight locations within Kachemak Bay, Alaska in 2018 and 2020 to examine spatial and temporal development of embryonic sporophytes on sori. Distinctions were made between sori collected from first-generation and overwintered adults to assess the influence of parental age on embryonic sporophyte release. Further distinctions were made between sori collected from attached and drifting individuals to assess the influence of the status of parental attachment to substrate on embryonic sporophyte release. Inspection of propagules released from sori after 48-h incubations indicated that embryonic sporophytes were occasionally released alongside viable spores. Though embryonic sporophytes were released from sori as early as spring, it was not evident that they were bound by seasonal or spatial limits. The percent of propagules that were embryonic sporophytes ranged from 0% to 100% but were not significantly different between first-generation and overwintered adults, nor were they different between attached and drifting individuals. Nevertheless, the characteristic of directly releasing embryonic sporophytes from adult sporophytes might have ecological advantages for N. luetkeana. Key words: Alaska; alternative; bull kelp; embryonic sporophytes; life history; Nereocystis luetkeana

Invaders as Diluents of the Cercarial Dermatitis Etiological Agent

Research on alien and invasive species focuses on the direct effects of invasion on native ecosystems, and the possible positive effects of their presence are most often overlooked. Our aim was to check the suitability of selected alien species (the snail Physa acuta, the bivalve Dreissena polymorpha, and the gammarid Dikerogammarus villosus) as diluents for infectious bird schistosome cercariae—the etiological factor of swimmer’s itch. It has been hypothesized that alien species with different feeding habits (scrapers, filterers and predators) that cohabit the aquatic environment with intermediate hosts of the schistosomatid trematodes are capable of feeding on their free-swimming stages—cercariae. In the laboratory conditions used, all experimental animals diluted the cercariae of bird schistosome. The most effective diluents were P. acuta and D. villosus. However, a wide discrepancy in the dilution of the cercariae between replicates was found for gammarids. The obtained results confirm the hypothesis that increased biodiversity, even when alien species are involved, creates the dilution effect of the free-living stages of parasites. Determining the best diluent for bird schistosome cercariae could greatly assist in the development of current bathing areas protection measures against swimmer’s itch.