The physical soldier caste of an invasive, human-infecting flatworm is morphologically extreme and obligately sterile.

We show that the globally invasive, human-infectious flatworm, Haplorchis pumilio, possesses the most physically specialized soldier caste yet documented in trematodes. Soldiers occur in colonies infecting the first intermediate host, the freshwater snail Melanoides tuberculata, and are readily distinguishable from immature and mature reproductive worms. Soldiers possess a pharynx five times absolutely larger than those of immature and mature reproductives, lack a germinal mass, and have a different developmental trajectory than reproductives, indicating that H. pumilio soldiers constitute a reproductively sterile physical caste. Neither immature nor mature reproductives showed aggression in in vitro trials, but soldiers readily attacked heterospecific trematodes that coinfect their host. Ecologically, we calculate that H. pumilio caused ~94% of the competitive deaths in the guild of trematodes infecting its host snail in its invasive range in southern California. Despite being a dominant competitor, H. pumilio soldiers did not attack conspecifics from other colonies. All prior reports documenting division of labor and a trematode soldier caste have involved soldiers that may be able to metamorphose to the reproductive stage and have been from nonhuman-infectious marine species; this study provides clear evidence for an obligately sterile trematode soldier, while extending the phenomenon of a trematode soldier caste to freshwater and to an invasive species of global public health concern.

Different metazoan parasites, different transcriptomic responses, with new insights on parasitic castration by digenetic trematodes in the schistosome vector snail Biomphalaria glabrata.

BACKGROUND: Gastropods of the genus Biomphalaria (Family Planorbidae) are exploited as vectors by Schistosoma mansoni, the most common causative agent of human intestinal schistosomiasis. Using improved genomic resources, overviews of how Biomphalaria responds to S. mansoni and other metazoan parasites can provide unique insights into the reproductive, immune, and other systems of invertebrate hosts, and their responses to parasite challenges. RESULTS: Using Illumina-based RNA-Seq, we compared the responses of iM line B. glabrata at 2, 8, and 40 days post-infection (dpi) to single infections with S. mansoni, Echinostoma paraensei (both digenetic trematodes) or Daubaylia potomaca (a nematode parasite of planorbid snails). Responses were compared to unexposed time-matched control snails. We observed: (1) each parasite provoked a distinctive response with a predominance of down-regulated snail genes at all time points following exposure to either trematode, and of up-regulated genes at 8 and especially 40dpi following nematode exposure; (2) At 2 and 8dpi with either trematode, several snail genes associated with gametogenesis (particularly spermatogenesis) were down-regulated. Regarding the phenomenon of trematode-mediated parasitic castration in molluscs, we define for the first time a complement of host genes that are targeted, as early as 2dpi when trematode larvae are still small; (3) Differential gene expression of snails with trematode infection at 40dpi, when snails were shedding cercariae, was unexpectedly modest and revealed down-regulation of genes involved in the production of egg mass proteins and peptide processing; and (4) surprisingly, D. potomaca provoked up-regulation at 40dpi of many of the reproduction-related snail genes noted to be down-regulated at 2 and 8dpi following trematode infection. Happening at a time when B. glabrata began to succumb to D. potomaca, we hypothesize this response represents an unexpected form of fecundity compensation. We also document expression patterns for other Biomphalaria gene families, including fibrinogen domain-containing proteins (FReDs), C-type lectins, G-protein coupled receptors, biomphalysins, and protease and protease inhibitors. CONCLUSIONS: Our study is relevant in identifying several genes involved in reproduction that are targeted by parasites in the vector snail B. glabrata and that might be amenable to manipulation to minimize their ability to serve as vectors of schistosomes.

Hooked on fish blood: the reliance of a gill parasite on haematophagy.

Parasitism involves diverse evolutionary strategies, including adaptations for blood feeding, which provides essential nutrients for growth and reproduction. Sparicotyle chrysophrii (Polyopisthocotyla: Microcotylidae), an ectoparasitic flatworm, infects the gills of gilthead seabream (Sparus aurata), significantly affecting fish health, welfare and Mediterranean cage farm profitability. Despite its impact, limited information exists on its feeding behaviour. This study demonstrates the presence of blood and exogenous haem groups in S. chrysophrii and explores its digestive tract using light and electron microscopy, elucidating its internal morphology and spatial arrangement. Elemental analysis of the digestive haematin cells shows residual oxidized haem depots as haematin crystals. Additionally, we studied the impact of the blood feeding on the host by estimating the average volume of blood intake for an adult parasite (2.84 ± 2.12µl·24h-1) and we described the significant drop of the plasmatic free iron levels in infected hosts. Overall, we demonstrate the parasite's reliance on its host blood, the parasite's buccal and digestive morphological adaptations for blood feeding and the provoked effect on the fish host's health.

Littoporins: Novel actinoporin-like proteins in caenogastropod genus Littorina.

In the course of searching for genes controlling the immune system in caenogastropod mollusks, we characterized and phylogenetically placed five new actinoporin-like cytolysins expressed in periwinkles of the genus Littorina. These newly discovered proteins, named littoporins (LitP), contain a central cytolysin/lectin domain and exhibit a predicted protein fold that is almost identical to the three-dimensional structures of actinoporins. Two of these proteins, LitP-1 and LitP-2, were found to be upregulated in L. littorea kidney tissues and immune cells in response to natural and experimental infection with the trematode Himasthla elongata, suggesting their potential role as perforins in the systemic anti-trematode immune response. The primary sequence divergence of littoporins is hypothesized to be attributed to the taxonomic range of cell membranes they can recognize and permeabilize.

Understanding the host response of farmed fish to blood flukes (Trematoda: Aporocotylidae) for developing new treatment strategies.

Aporocotylids (Trematoda: Digenea), also known as fish blood flukes infect the circulatory system of fish leading to serious health problems and mortality. Aporocotylids are a particular concern for farmed fish as infection intensity can increase within the farming environment and lead to mortalities. In the context of managing these infections, one of the most crucial aspects to consider is the host response of the infected fish against these blood flukes. Understanding the response is essential to improving current treatment strategies that are largely based on the use of anthelmintic praziquantel to manage infections in aquaculture. This review focuses on the current knowledge of farmed fish host responses against the different life stages of aporocotylids. New treatment strategies that are able to provide protection against reinfections should be a long-term goal and is not possible without understanding the fish response to infection and the interactions between host and parasite.

Behave yourself: effects of exogenous-glucocorticoid exposure on larval amphibian anti-parasite behaviour and physiology.

Parasites represent a ubiquitous threat for most organisms, requiring potential hosts to invest in a range of strategies to defend against infection-these include both behavioural and physiological mechanisms. Avoidance is an essential first line of defence, but this behaviour may show a trade-off with host investment in physiological immunity. Importantly, while environmental stressors can lead to elevated hormones in vertebrates, such as glucocorticoids, that can reduce physiological immunity in certain contexts, behavioural defences may also be compromised. Here, we investigate anti-parasite behaviour and immune responses against a trematode (flatworm) parasite by larval amphibians (tadpoles) exposed or not to a simulated general stressor in the form of exogenous corticosterone. Tadpoles that were highly active in the presence of the trematode infectious stage (cercariae) had lower infection loads, and parasite loads from tadpoles treated only with dechlorinated water were significantly lower than those exposed to corticosterone or the solvent control. However, treatment did not affect immunity as measured through white blood-cell profiles, and there was no relationship between the latter and anti-parasite behaviour. Our results suggest that a broad range of stressors could increase host susceptibility to infection through altered anti-parasite behaviours if they elevate endogenous glucocorticoids, irrespective of physiological immunity effects. How hosts defend themselves against parasitism in the context of multiple challenges represents an important topic for future research, particularly as the risk posed by infectious diseases is predicted to increase in response to ongoing environmental change.

Ecological consequences of hidden pathology by larval digeneans in South American mollusks.

The study of digeneans and their association with mollusks commenced in Europe and South America during the mid-19th to early 20th centuries. Digenean infestation can severely degrade host tissue, leading to diminished energy resources and eventual host mortality. However, these parasites can also induce various non-lethal effects, including changes in growth rates, survival rates, and reproductive capabilities, alongside physiological and behavioral alterations. While numerous studies have explored the ecological effects of digeneans on hosts in Europe and North America, our understanding of these dynamics in South America, particularly in first intermediate hosts, remains limited. Therefore, this paper aims to provide an overview of ecological investigations into digenean-mollusk systems in South America, emphasizing the importance of robust sampling designs and statistical analyses to address key ecological inquiries. Although fascinating examples exist of parasitism influencing different hierarchical levels of digenean-mollusk systems, particularly at the individual, population, and community levels, documentation of their ecosystem-level impacts is comparatively sparse. As South American studies of digenean-mollusk systems from an ecological perspective are still in their early stages, there is immense potential for uncovering unique ecological patterns in this largely unexplored region, propelling us toward further developmental strides in the parasite ecology.

Characterisation of Nematoda and Digenea in selected Australian freshwater snails.

Freshwater snails are integral to local ecosystems as a primary food source for various vertebrate species, thereby contributing significantly to ecological food webs. However, their role as intermediate hosts also makes them pivotal in the transmission of parasites. In Australia, research on freshwater snails has predominantly focused on their role as intermediate hosts for livestock parasites, while there has been limited exploration of the impact of these parasites on snail health and population dynamics. The aim of this study was to determine parasitic infection in freshwater snails. This study was conducted in the south-eastern region of Australia, in 2022. A total of 163 freshwater snails from four different species were collected and examined in the Murrumbidgee catchment area in the southeastern part of Australia during the Southern Hemisphere summer and autumn months (February to May). The species included Isidorella hainesii, Glyptophysa novaehollandica, Bullastra lessoni (endemic species), and Physella acuta (an introduced species). Through the analysis of sequence data from the various regions of the nuclear ribosomal DNA, we determined that the Digenea species in this study belonged to three distinct species, including Choanocotyle hobbsi, Petasiger sp. and an unidentified species belonging to Plagiorchioidea. Additionally, analysis of the sequences from Nematoda found in this study, revealed they could be categorized into two separate taxa, including Krefftascaris sp. and an unidentified nematode closely associated with plant and soil nematodes. This research holds significant implications for the future understanding and conservation of Australian freshwater ecosystems. Most parasites found in the present study complete their life cycle in snails and turtles. As many of freshwater snail and turtle species in Australia are endemic and face population threats, exploring the potential adverse impacts of parasitic infections on snail and turtle health, is crucial for advancing our understanding of these ecosystems and also paving the way for future research and conservation efforts. While none of the native snail species in the present study have been listed as endangered or threatened, this may simply be attributed to the absence of regular population surveys.

Morphological and molecular characterization of brown-banded broodsacs and metacercariae of Leucochloridium (Trematoda: Leucochloridiidae) parasitizing the semi-slug Omalonyx unguis (Succineidae) in Argentina.

Trematodes of the genus Leucochloridium exhibit an unusual transmission strategy among mollusks (intermediate host). The fully developed sporocyst, housing encysted metacercariae, displays vivid coloration and rhythmic activity in the snail's tentacle, mimicking insect larvae. These strategies attract insectivorous birds, their final hosts, thereby increasing the chances of completing their life cycle. In South America, the reports of adults and larval stages of Leucochloridium are scarce. Brown-banded broodsac of Leucochloridium sp. were obtained from Omalonyx unguis collected in a shallow lake from Corrientes Province, Argentina. Here, we morphologically characterized the larval stages (broodsac and metacercaria), identified the parasite through DNA sequences from nuclear 28S-rRNA (28S) and the mitochondrial cytochrome c oxidase I (COI) genes, and explored its evolutionary affinities with the Leucochloridium species available in GenBank. The present broodsac displays brown bands, with a yellowish background in the first two-thirds and yellowish-white in the last third. Based on morphological comparisons, the broodsac and metacercaria described in this study could not be conclusively categorized under any known South American species of Leucochloridium. In relation to the phylogenetic reconstructions, Leucochloridium sp. consistently clustered with L. perturbatum, and species delimitation analyses resulted in recognized Leucochloridium sp. from Argentina as a distinct species. The DNA sequences obtained in this study constitute the first genetic data generated for sporocyst broodsacs in South America. Future studies, incorporating morphology, genetic, and biological data, will be essential for both species identification and the elucidation of leucochloridiid diversity in the region.

Form and Function in the Digenea, with an Emphasis on Host-Parasite and Parasite-Bacteria Interactions.

This review covers the general aspects of the anatomy and physiology of the major body systems in digenetic trematodes, with an emphasis on new knowledge of the area acquired since the publication of the second edition of this book in 2019. In addition to reporting on key recent advances in the morphology and physiology of tegumentary, sensory, neuromuscular, digestive, excretory, and reproductive systems, and their roles in host-parasite interactions, this edition includes a section discussing the known and putative roles of bacteria in digenean biology and physiology. Furthermore, a brief discussion of current trends in the development of novel treatment and control strategies based on a better understanding of the trematode body systems and associated bacteria is provided.

Other Schistosomatoidea and Diplostomoidea.

Trematodes of the order Diplostomida are well known as serious pathogens of man, and both farm and wild animals; members of the genus Schistosoma (Schistosomatidae) are responsible for human schistosomosis (schistosomiasis) affecting more than 200 million people in tropical and subtropical countries, and infections of mammals and birds by animal schistosomes are of great veterinary importance. The order Diplostomida is also rich in species parasitizing other major taxa of vertebrates. The "Aporocotylidae" sensu lato are pathogenic in fish, "Spirorchiidae" sensu lato in reptiles. All these flukes have two-host life cycles, with asexually reproducing larvae usually in mollusks and occasionally in annelids, and adults usually live in the blood vessels of their vertebrate hosts. Pathology is frequently associated with inflammatory reactions to eggs trapped in various tissues/organs. On the other hand, the representatives of Diplostomidae and Strigeidae have three- or four-host life cycles in which vertebrates often serve not only as definitive but also as intermediate or paratenic hosts. Pathology is usually associated with migration of metacercariae and mesocercariae within the host tissues. The impact of these trematode infections on both farm and wild animals may be significant.

Amphistomes.

Amphistomes, commonly referred to as 'stomach' or 'rumen' flukes because of the localization of these flukes in the stomach of ruminants, are digenetic trematodes distinguished by the absence of an oral sucker and the position of the ventral sucker or acetabulum at the posterior end of the body. The body is characterized by a leaf-like fleshy structure, pink or red with a large posterior sucker. Amphistomes are an important group of parasites since they cause 'amphistomiasis' (variously known as paramphistomosis/amphistomosis), a serious disease of great economic importance in ruminants worldwide. These parasites have a broad spectrum of definitive hosts together with a wide geographical distribution. Though they form a continuous evolutional lineage from fishes to mammals, amphistomes mainly inhabit the rumen and reticulum of ruminant mammals, while some species occur in the large intestine or parenteric sites of ruminants, pigs, equines and man.

Selected Wildlife Trematodes.

The trematodes are a species-rich group of parasites, with some estimates suggesting that there are more than 24,000 species. However, the complexities associated with their taxonomic status and nomenclature can hinder explorations of the biology of wildlife trematodes, including fundamental aspects such as host use, life cycle variation, pathology, and disease. In this chapter, we review work on selected trematodes of amphibians, birds, mammals, and their snail intermediate hosts, with the goal of providing a tool kit on how to study trematodes of wildlife. We provide a brief introduction to each group of wildlife trematodes, followed by some examples of the challenges each group of trematodes has relative to the goal of their identification and understanding of the biology and interactions these organisms have with their wildlife hosts.

Echinostomes and Other Intestinal Trematode Infections.

Intestinal trematodes are among the most common types of parasitic worms. About 76 species belonging to 14 families have been recorded infecting humans. Infection commonly occurs when humans eat raw or undercooked foods that contain the infective metacercariae. These parasites are diverse in regard to their morphology, geographical distribution and life cycle, which make it difficult to study the parasitic diseases that they cause. Many of these intestinal trematodes have been considered as endemic parasites in the past. However, the geographical limits and the population at risk are currently expanding and changing in relation to factors such as growing international markets, improved transportation systems, new eating habits in developed countries and demographic changes. These factors make it necessary to better understand intestinal trematode infections. This chapter describes the main features of human intestinal trematodes in relation to their biology, epidemiology, host-parasite relationships, pathogenicity, clinical aspects, diagnosis, treatment and control.

Effect of trematode infection on cardiac activity of marine gastropod Littorina littorea in situ and under laboratory conditions.

Physiological mechanisms underlying relationships between the trematode parthenitae and their molluscan hosts are poorly understood. In this study, we estimated the cardiac function of gastropods Littorina littorea L. infected with Himasthla elongata and Cryptocotyle lingua under laboratory conditions and in situ. The heart rate (HR) of the infected periwinkles was significantly lower than in the uninfected ones. HR of uninfected periwinkles in the natural environment followed a clear tidal pattern, with high values at high tide and low values at low tide. However, HR in most of the infected molluscs did not follow this rhythm. The decrease in HR was more pronounced in the periwinkles infected with C. lingua than with H. elongata. Thus, infection with C. lingua had a greater effect on the physiology and metabolic rate of periwinkles than infection with H. elongata.

Littorina snails and Microphallus trematodes: Diverse consequences of the trematode-induced metabolic shifts.

The intricate relationships between parasites and hosts encompass a wide range of levels, from molecular interactions to population dynamics. Parasites influence not only the physiological processes in the host organism, but also the entire ecosystem, affecting mortality of individuals, the number of offspring through parasitic castration, and matter and energy cycles. Understanding the molecular mechanisms that govern host-parasite relationships and their impact on host physiology and environment remains challenging. In this study, we analyzed how infection with Microphallus trematodes affects the metabolome of two Littorina snail species inhabiting different intertidal zone shore levels. We applied non-targeted GC-MS-based metabolomics to analyze biochemical shifts induced by trematode infection in a host organism. We have identified changes in energy, amino acid, sugar, and lipid metabolism. In particular, we observed intensified amino acid catabolism and nitrogenous catabolites (glutamine, urea) production. These changes primarily correlated with infection and interspecies differences of the hosts rather than shore level. The changes detected in the host metabolism indicate that other aspects of life may have been affected, both within the host organism and at a supra-organismal level. Therefore, we explored changes in microbiota composition, deviations in the host molluscs behavior, and acetylcholinesterase activity (ACE, an enzyme involved in neuromuscular transmission) in relation to infection. Infected snails displayed changes in their microbiome composition. Decreased ACE activity in snails was associated with reduced mobility, but whether it is associated with trematode infection remains unclear. The authors suggest a connection between the identified biochemical changes and the deformation of the shell of molluscs, changes in their behavior, and the associated microbiome. The role of parasitic systems formed by microphallid trematodes and Littorina snails in the nitrogen cycle at the ecosystem level is also assumed.

Season, size, and sex: factors influencing monogenean prevalence and intensity on Gambusia affinis in New Zealand.

A number of studies have been conducted on monogenean seasonality, though primarily in continental regions with wide annual temperatures ranges. We investigated seasonal changes in the prevalence and intensity of Salsuginus seculus infesting sexually dimorphic western mosquitofish (Gambusia affinis) in New Zealand. This represents the first examination of seasonality for this species globally, and the first seasonal assessment of any monogenean population in New Zealand, a temperate country with a mild oceanic climate. Prevalence and intensity of S. seculus with respect to fish size and sex was also examined. Prevalence of S. seculus changed temporally, peaking in summer, and was strongly positively correlated with algal concentrations. This relationship may be associated with increasing food levels, leading to an increase in fish courting and mating, resulting in high numbers and close physical associations of G. affinis individuals, facilitating transmission of the monogeneans. Thus, biotic factors may be important in determining temporal changes in S. seculus prevalence in New Zealand. Female G. affinis had a significantly higher prevalence and mean intensity of S. seculus than males. Longer fish had a higher mean intensity and prevalence of S. seculus. Female G. affinis likely host disproportionately more monogeneans as they are larger than males. Alternatively, females may have a compromised immune response during reproductive periods. Overall, seasonal change was observed in S. seculus prevalence and intensity under New Zealand's mild climatic conditions, and the larger female G. affinis in this dimorphic species supported a greater prevalence and intensity of infestation than males.

Parasite abundance distribution as a model of host-parasite relationships between monogeneans Gyrodactylus spp. and cage-reared rainbow trout Oncorhynchus mykiss.

Aggregation is a fundamental feature of parasite distribution in the host population, but the biological implications of the aggregation indices used to describe the relationships between the populations of parasites and hosts are not evident. It is speculated that the form of distribution in each case is predicated on the host's varying resistance to the infection, which is hard to control, making it difficult to adequately interpret the index values. This paper examines several cases from trout farms in Russian Karelia to explore the monogenean Gyrodactylus spp. infection in rainbow trout of varying ages. The genetic homogeneity of cage-reared fish and the direct life cycle of the helminths make the relationship between the species more lucid than in natural host-parasite systems. The results give no ground to speak of any specific patterns: as well as in the natural systems, the infection rates in trout vary widely, i.e., the helminth distribution has not become more uniform; the observed distributions in all cases are adequately approximated by the negative binomial model; the positive abundance-occupancy relationships (AORs) and abundance-variance relationships (AVRs) common for parasitic systems apply to the basic infection parameters. The form of the negative binomial distribution is shaped by two parameters-k and θ, the former being a metric of the infection variability, which depends on the host's individual resistance, and the latter representing the parasites' reproduction and establishment success rates. A rise in the parameter k indicates increased aggregation and a higher parameter θ points to a more uniform frequency distribution. These parameters can be used as a representative tool for monitoring the parasite communities in salmonid fishes, including in aquaculture.

Gyrodactylus in the spotlight: how exposure to light impacts disease and the feeding behavior of the freshwater tropical guppy (Poecilia reticulata).

Artificial light at night (ALAN) negatively impacts organisms in many ways, from their feeding behaviors to their response and ability to deal with disease. Our knowledge of ALAN is focused on hosts, but we must also consider their parasites, which constitute half of all described animal species. Here, we assessed the impact of light exposure on a model host-parasite system (Poecilia reticulata and the ectoparasitic monogenean Gyrodactylus turnbulli). First, parasite-free fish were exposed to 12:12 h light:dark (control) or 24:0 h light:dark (ALAN) for 21 days followed by experimental infection. Second, naturally acquired G. turnbulli infections were monitored for 28 days during exposure of their hosts to a specified light regime (6:18 h, 12:12 h, or 24:0 h light:dark). Experimentally infected fish exposed to constant light had, on average, a greater maximum parasite burden than controls, but no other measured parasite metrics were impacted. Host feeding behavior was also significantly affected: fish under ALAN fed faster and took more bites than controls, whilst fish exposed to reduced light fed slower. Thus, ALAN can impact parasite burdens, even in the short term, and altering light conditions will impact fish feeding behavior. Such responses could initiate disease outbreaks or perturb food-webs with wider ecological impacts.

Manipulative neuroparasites: uncovering the intricacies of neurological host control.

Manipulative neuroparasites are a fascinating group of organisms that possess the ability to hijack the nervous systems of their hosts, manipulating their behavior in order to enhance their own survival and reproductive success. This review provides an overview of the different strategies employed by manipulative neuroparasites, ranging from viruses to parasitic worms and fungi. By examining specific examples, such as Toxoplasma gondii, Leucochloridium paradoxum, and Ophiocordyceps unilateralis, we highlight the complex mechanisms employed by these parasites to manipulate their hosts' behavior. We explore the mechanisms through which these parasites alter the neural processes and behavior of their hosts, including the modulation of neurotransmitters, hormonal pathways, and neural circuits. This review focuses less on the diseases that neuroparasites induce and more on the process of their neurological manipulation. We also investigate the fundamental mechanisms of host manipulation in the developing field of neuroparasitology, which blends neuroscience and parasitology. Finally, understanding the complex interaction between manipulative neuroparasites and their hosts may help us to better understand the fundamentals of behavior, neurology, and host-parasite relationships.