Spread of An Avian Eye Fluke, Philophthalmus gralli, Through Biological Invasion of An Intermediate Host.

Philophthalmus is a genus of globally distributed parasitic eye flukes with some members of the genus found in disparate locales. In particular, Philophthalmus gralli, a zoonotic trematode, appears to be a relatively new introduction to the Americas, facilitated by spillover from the invasive snails Melanoides tuberculata (red-rimmed melania) and Tarebia granifera (quilted melania), which were introduced via the aquarium trade, and perhaps furthered by avian dispersal. Given that two known intermediate hosts of Philophthalmus flukes are actively expanding their range as a result of human activities, we hypothesize that this spread is also associated with the spread of Philophthalmus flukes. To address this, we systematically reviewed the literature and examined whether the global expansion of P. gralli flukes is associated with the spread of invasive snails M. tuberculata and T. granifera. Here, we show that (1) specimens of P. gralli are only found in intermediate snail hosts M. tuberculata or T. granifera, suggesting intermediate host specificity for these 2 species, and (2) specimens of P. gralli have rarely been found outside the ranges (native and introduced) of M. tuberculata or T. granifera. Given the importance of distribution information of parasites in the role of identifying parasite invasions, we also review the known distribution of all Philophthalmus species. Considering recent outbreaks in humans and wild and domestic animal species, the continued spread of Philophthalmus presents a potential threat to veterinary and public health and conservation.

Timing and order of exposure to two echinostome species affect patterns of infection in larval amphibians.

The study of priority effects with respect to coinfections is still in its infancy. Moreover, existing coinfection studies typically focus on infection outcomes associated with exposure to distinct sets of parasite species, despite that functionally and morphologically similar parasite species commonly coexist in nature. Therefore, it is important to understand how interactions between similar parasites influence infection outcomes. Surveys at seven ponds in northwest Pennsylvania found that multiple species of echinostomes commonly co-occur. Using a larval anuran host (Rana pipiens) and the two most commonly identified echinostome species from our field surveys (Echinostoma trivolvis and Echinoparyphium lineage 3), we examined how species composition and timing of exposure affect patterns of infection. When tadpoles were exposed to both parasites simultaneously, infection loads were higher than when exposed to Echinoparyphium alone but similar to being exposed to Echinostoma alone. When tadpoles were sequentially exposed to the parasite species, tadpoles first exposed to Echinoparyphium had 23% lower infection loads than tadpoles first exposed to Echinostoma. These findings demonstrate that exposure timing and order, even with similar parasites, can influence coinfection outcomes, and emphasize the importance of using molecular methods to identify parasites for ecological studies.

The effect of trematode infection on the markers of oxidative stress in the offspring of the freshwater snail Lymnaea stagnalis.

Most invertebrate species exhibit immunological responses that can inactivate and eliminate penetrating parasites. Such immune responses in particular involve the formation of potentially toxic reactive oxygen species (ROS). We explored the immune capabilities of the first-generation (F1) offspring of naturally infected freshwater snails, Lymnaea stagnalis, in response to infection by trematode cercariae under laboratory conditions. The rates of ROS formation and peroxidase activity in the hemolymph of the F1 offspring of L. stagnalis parents infected by an asexual stage of trematodes were significantly higher than in F1 offspring of uninfected parents. Compared to offspring from uninfected parents, the growth rate of F1 snails from infected parents was higher, but survival was lower. After infection of F1 snails by trematode cercariae of Echinoparyphium aconiatum under laboratory conditions, the rate of ROS formation and peroxidase activity in the hemolymph of F1 offspring of uninfected parents increased compared to control snails. This pattern persisted throughout the entire 3-week observation period. In contrast, the rate of ROS formation in the hemolymph of F1 snails from infected parents after experimental infection by E. aconiatum cercariae did not differ from controls, and peroxidase activity even decreased. Thus, trematode parthenitae infection of parents could alter the immune response of their offspring.

First molecular identification and phylogenetic tree of Petasiger exaeretus Dietz, 1909 (Digenea: Echinostomatidae) from an intermediate host Radix auricularia (L., 1758) in Greater Zab river, Iraq.

INTRODUCTION: Radix auricularia (Linnaeus, 1758) is a freshwater gastropod belongs to the Lymnaeidae (pond snails) family which act as intermediate hosts or vectors of various parasitic flukes. No study has yet been undertaken on the prevalence of Petasiger spp. infection in R. auricularia. Species of Petasiger (Dietz, 1909) are a cosmopolitan parasite that utilize snails as the first intermediate host, with vertebrates like amphibians larvae and fish as the second intermediate host, followed by fish-eating birds. The current paper is considered to be the first report of Petasiger exaeretus parasitized R. auricularia in Iraq, which is supported with molecular and phylogenetic analysis. MATERIAL AND METHODS: Freshwater snails R. auricularia were collected during October 2016 - September 2017 from different locations of Sufaia village on the Greater Zab river, Erbil province, Iraq. RESULTS: A total of 307 freshwater snails R. auricularia were collected, only five of them were infected with a prevalence of Petasiger exaeretus (1.62%). CONCLUSIONS: The current study agrees with the opinion of Selbach, Soldánová (26), which suggested the possibility of a much higher morphological diversity within Petasiger species, based on the number of described cercariae, compared with adult forms.It is clear that P. phalacrocoracis specimens have often been erroneously designated as P. exaeretus by many authors (Nasincová et al., 1994). Certain morphological similarities and dissimilarities between P. exaeretus and P. phalacrocoracis can be detected: the pear-shaped body resembles P. exaeretus, whereas, P. phalacrocoracis have an elongated body.

Molecular biological studies of adult and metacercarial stages of Petasiger exaeretus Dietz, 1909 (Digenea: Echinostomatidae).

Molnár et al. (2015) reported two types of echinostomatid metacercariae in the lateral line organ of Hungarian fish species. Type 1 metacercariae possessed 27 collar spines and 16 uniform and three larger dorsal spines, whereas Type 2 metacercariae bore 27 collar spines and 19 equal-sized dorsal spines. In the recent work, molecular studies carried out on the ITS region and partial 28S rDNA sequences of two types of echinostomatid metacercariae and the sequences of adult stages of the species of Petasiger Dietz, 1909 collected from cormorants (Phalacrocorax carbo L.) showed that some of the Type 2 metacercariae corresponded to Petasiger exaeretus Dietz, 1909, whereas other morphologically similar metacercariae were identified as Petasiger phalacrocoracis (Yamaguti, 1939). The sequences of the Type 1 metacercariae with three larger dorsal spines could not be identified with any of the known sequences from echinostomatid trematodes.

Immediate and lag effects of pesticide exposure on parasite resistance in larval amphibians.

Across host-parasite systems, there is evidence that pesticide exposure increases parasite loads and mortality following infection. However, whether these effects are driven by reductions in host resistance to infection or slower rates of parasite clearance is often unclear. Using controlled laboratory experiments, we examined the ability of larval northern leopard frogs (Lithobates pipiens) and American toads (Anaxyrus americanus) to resist and clear trematode (Echinoparyphium sp.) infections following exposure to the insecticide carbaryl. Northern leopard frogs exposed to 1 mg L-1 of carbaryl had 61% higher parasite loads compared with unexposed individuals, while there was no immediate effect of carbaryl on parasite encystment in American toads. However, when tadpoles were exposed to carbaryl and moved to freshwater for 14 days before the parasite challenge, we recovered 37 and 63% more parasites from carbaryl-exposed northern leopard frogs and American toads, respectively, compared with the control. No effects on clearance were found for either species. Collectively, our results suggest that pesticide exposure can reduce the ability of amphibians to resist parasite infections and that these effects can persist weeks following exposure. It is critical for researchers to incorporate species interactions into toxicity studies to improve our understanding of how contaminants affect ecological communities.

Trematodes with a reproductive division of labour: heterophyids also have a soldier caste and early infections reveal how colonies become structured.

Recent findings have extended the documentation of complex sociality to the Platyhelminthes, describing the existence of a reproductive division of labour involving a soldier caste among the parthenitae of trematode parasites. However, all species examined to date occupy high positions in trematode interspecific dominance hierarchies and belong to two closely related families, the Echinostomatidae and the Philophthalmidae (Superfamily Echinostomatoidea). Further, the two species documented as lacking soldiers also belong to the Echinostomatidae. Here, we examine four species of intermediate dominance, all belonging to the family Heterophyidae (Superfamily Opisthorchioidea): Euhaplorchis californiensis, Phocitremoides ovale, Pygidiopsoides spindalis and Stictodora hancocki, all of which infect the California horn snail, Cerithideopsis californica (=Cerithidea californica). We quantify morphology, distribution and behaviour of rediae from fully developed colonies. We also provide information on colony structure for three developing heterophyid colonies to better understand colony development. We discuss the implications of our findings, particularly with respect to how they suggest alternatives to the conclusions of other researchers concerning the nature of trematode sociality. Our analyses of morphological, distributional and behavioural patterns of developed colonies indicate that these heterophyid trematodes have a non-reproductive caste whose function is defence of the colony from invading trematodes. Hence, a soldier caste occurs for species lower in dominance hierarchies than previously known, and is present in at least two superfamilies of digenean trematodes, suggesting that selection for a soldier caste may be much more common among the Trematoda than previously recognised.

Malformations of the gill filaments of the ruffe Gymnocephalus cernuus (L.) (Pisces) caused by echinostomatid metacercariae.

In parasite surveys of fishes from Lake Balaton and its tributaries in Hungary, infections with metacercariae of a species of the digenean genus Echinochasmus (Trematoda: Echinostomatidae) were found in seven species of fish. In ruffe, Gymnocephalus cernuus, malformations of the gill filaments apparently caused by these infections were observed. These malformations were in the form of bifurcations of the filaments at about their mid-length. At the point where the filaments bifurcate, an Echinochasmus metacercaria was always embedded in the cartilaginous ray of the gill filament. All specimens of the ruffe were found to be infected by these metacercariae, and each ruffe specimen was infected by 30-300 metacercariae. Such a bifurcation was found in all of the ruffe specimens, but, apart from these gill malformations, the metacercariae produced only local changes in the cartilage. In the other six infected fish species, only local signs were observed in the cartilage. Experimental infections of chicks with metacercariae resulted in the finding of the sexual adult (marita) of an unidentified species of Echinochasmus. ITS sequences of the adult and metacercaria corresponded with each other, and also with a cercaria isolated from a gravel snail (Lithoglyphus naticoides), with a 99.5-100% similarity.

The influence of trematode metacercariae on the individual fecundity of Bithynia troscheli (Gastropnoda: Bithyniidlae).

Fifteen trematode species belonging to seven families parasitizing the females of Bithynia troscheli (Paasch, 1842) at metacercarial stage, were recorded in Kargat River (Lake Chany, South of West Siberia, Russia). The breeding mollusk females have less metacercarial diversity (9 species vs. 15) and lower mean parasite abundance, comparing to non-ovigerous mollusks. The hypothesis of the metacercarial influence on individual fecundity of aquatic gastropods was tested. The individual fecundit pariameters of the-B. troscheli females, uninfected and infected with the trematode metacercariae belonging to fam. Echinostomatidae, Cyathocotilidae, Strigeidae, Cyclocoelidae, were compared. The percentage of the breeding B. troscheli females--was less and their fecundity parameters were lower for the hosts infected (with the Strigeidae or Cyclocoelidae metacercariae) comparing to non-infected ones. Trematode metacercariae found in the mantle or somatic musculature had no significant effects on the fecundity of the host. However, our results showed that trematode metacercariae affect the reproduction of B. troscheli in dual ways. All fecundity parameters of the mollusk females with the high infection rate were, significantly lower than those for uninfected females: the percentage of fecund females (chi2 = 6.73 ,p < 0.01), number of clutches per female, number of egg capsules per clutch and number of normal egg capsules per female (Tukey HSD, p < 0.001). Although the females with-low intensity of metacercarial infection lay egg clutches significantly more frequently than uninfected ones (chi2 = = 4.18, P = 0.04); the fecundity parameters were approximately equal for both groups of the mollusk females. These pioneer results prove the reality of influence of metacercariae on the individual fecundity of aquatic gastropods. The regulatory population mechanisms that may compensate the host reproduction loss caused by metacercaria, are discussed.

Acanthoparyphium sp. and other metazoan symbionts of the American oyster, Crassostrea virginica, from South Texas.

In this study, we investigated oysters, Crassostrea virginica , from Laguna Madre in South Texas, where a 45-yr old study recorded metacercarial infections of the echinostomatid trematode, Acanthoparyphium spinulosum , an Asian relative of which, Acanthoparyphium tyosenense, has been associated with human infections via the ingestion of raw mollusks. In an effort to examine the base-line infection parameters of Acanthoparyphium sp. in oysters, we examined the effect of distance from the shoreline, which is the habitat of the first intermediate host snail, Cerithidea pliculosa, as well as temporal changes in oyster infection levels, by conducting quarterly collections of oysters during a year. We found that almost all oysters (prevalence = 97.8-100%) were infected regardless of distance to the shoreline and season. However, the abundance of metacercariae was significantly higher close to the shoreline, while no significant temporal changes could be detected. In addition to the echinostomatid, we found a high abundance of the metacestode Tylocephalum sp. and the presence of 4 other metazoan parasites. None of the infections seemed to incur significant tissue damage to the oysters. Our study shows that at least locally, recreational harvesters of oysters may be exposed to Acanthoparyphium sp. Future studies should examine oysters from snail host habitats in the Gulf of Mexico, and the potential zoonotic risk of Acanthoparyphium sp. should be evaluated using experimental infections of animal models.

Behavioural plasticity of social trematodes depends upon social context.

Members of some social insects adjust their behaviours depending upon social context. Such plasticity allows colonies to sustain efficiency of the whole without the cost of additional production of individuals or delayed responses to perturbations. Using the recently discovered social clonal stage of trematode parasites, we investigated whether members of the reproductive caste adjust their defensive behaviour according to the local availability of non-reproductive defensive specialists, and if so whether the plasticity affects the short-term reproductive success of reproductive morphs. In vitro experiments demonstrated plasticity in competitive interactions of the reproductive morphs depending on the number of non-reproductive defensive specialists present nearby, which lead to differences in reproductive output at the individual level. This study provides support for the benefit of maintaining non-reproductive morphs in competitive situations, arising through socially mediated behavioural plasticity.

[Clonal variability in longevity of the cercariae of Himasthla elongata (Trematoda: Echinostomatidae)].

The study was carried out on Himasthla elongata cercariae shed by infected Littorina littorea snails. The infected periwinkles were collected from the settlement with the low prevalence of H. elongata. As shown earlier with the use of AFLP (Amplified Fragment Length Polymorphisms) method, rediae groups in all the infected periwinkles of this settlement arise from the infection of a mollusc with a single miracidium. Therefore, the cercariae shed by an infected mollusc have the same genotype or, in other words, represent a clone. The LT50 (the time during which 50% of cercariae perish in the experimental dish) were measured experimentally for cercariae Himasthla elongata belong to different clones. The investigated parameter demonstrated a high level of interclonal variability. Two groups of cercarial clones were identified: one of them was characterized by the high level of intraclonal variability in LT50 and the second, by the low one. It is assumed that the observed heterogeneity may be stipulated by different degrees of mitotic recombinations during formation of different cercarial clones.

The biology of Echinoparyphium (Trematoda, Echinostomatidae).

Echinoparyphium species are common, widely distributed intestinal parasites causing disease in animals worldwide. Intermediate hosts include snails, bivalves, and fish, whereas the definitive hosts are mainly birds and mammals. This review examines the significant literature on Echinoparyphium. Descriptive studies, life cycle, experimental and manipulative studies, and biochemical and molecular studies are presented. The influence of environmental factors, and toxic pollutants, are reviewed as well as studies on the pathology of Echinoparyphium.

Host quality and spatial patterning in infections of the Eastern mudsnail (Ilyanassa obsoleta) by two trematodes (Himasthla quissetensis and Zoogonus rubellus).

Several studies have suggested that the fitness of a parasite can be directly impacted by the quality of its host. In such cases, selective pressures could act to funnel parasites towards the highest-quality hosts in a population. The results of this study demonstrate that snail host quality is strongly correlated with spatial patterning in trematode infections and that habitat type is the underlying driver for both of these variables. Two trematodes (Himasthla quissetensis and Zoogonus rubellus) with very different life cycles assume the same spatial infection pattern in populations of the first intermediate host (Ilyanassa obsoleta) in coastal marsh habitats. Infected snails are disproportionately recovered from intertidal panne habitats, which offer more hospitable environs for snails than do adjacent habitats (intertidal creeks, coastal flats, and subtidal creeks), in terms of protection from turbulence and wave action, as well as the availability of food stuffs. Snails in intertidal panne habitats are of higher quality when assessed in terms of average size-specific mass, growth rate, and fecundity. In mark-recapture experiments, snails frequently dispersed into intertidal pannes but were never observed leaving them. In addition, field experiments demonstrate that snails confined to intertidal panne habitats are disproportionately infected by both trematode species, relative to conspecifics confined to adjacent habitats. Laboratory experiments show that infected snails suffer significant energetic losses and consume more than uninfected conspecifics, suggesting that infected snails in intertidal pannes may survive better than in adjacent habitats. We speculate that 1 possible mechanism for the observed patterns is that the life cycles of both trematode species allows them to contact the highest-quality snails in this marsh ecosystem.

In vivo efficacy of praziquantel against Echinoparyphium aconiatum (Trematoda: Echinostomatidae) parasitizing the great pond snails Lymnaea stagnalis (Gastropoda: Lymnaeidae).

The present study had a practical goal. I aimed to determine whether praziquantel could reduce the production of Echinoparyphium aconiatum (Trematoda: Echinostomatidae) cercariae in infected snails Lymnaea stagnalis (Gastropoda: Lymnaeidae) without killing the hosts. Praziquantel is a broad-spectrum antihelminth agent. It caused a total cessation of cercaria shedding when the praziquantel concentration in the treatment bath was 10 mg/L and the treatment time was 30 h or longer. A next research step which has to be taken before giving detailed recommendations about using praziquantel for ceasing production of E. aconiatum cercariae in parasitized snails is to follow the survivorship and performance of treated snails after a praziquantel exposure for longer than in this medium-term (3 days) experiment.

[Artyfechinostomum sufratyfex Lane, 1915 (Trematoda:Echinostomatidae), a parasite of mammals in Southern Vietnam].

Trematoda Artyfechinostomumn sufratyfex Lane, 1915 was found in different mammal species of Southern Vietnam. Original measurements of this species from domestic pig, black rat, and Asian palm civet are given.

Scanning electron microscopy of adult Echinoparyphium recurvatum (Von Linstow, 1873) (Digenea: Echinostomatidae) from Britain.

The surface topography of 15-day-old adult Echinoparyphium recurvatum (Von Linstow, 1873) sensu stricto, from an isolate of the parasite utilizing Lymnaea peregra as first intermediate host in southern England, is described and illustrated using scanning electron microscopy, and is compared to those of other Echinoparyphium species from Europe, and with those of E. recurvatum of East Asian origin. The general morphology of tegumental features was found to be very similar to that of worms of the same age observed in a previous study on a Korean isolate of E. recurvatum. Comparison of collar and body spination of E. recurvatum with other 45-collar-spined members of the genus in Europe revealed some significant differences in morphology. Collar spines of E. recurvatum were found to be shorter and more pointed than those of Echinoparyphium mordwilkoi. The body spines of E. recurvatum are rounded and scale-like, extending just beyond the ventral sucker, contrasting with the pointed, thorn-shaped body spines of E. mordwilkoi, extending posteriorly to the level of the second testis. Body spine shape and distribution in E. recurvatum were found to be more similar to those of Echinoparyphium pseudorecurvatum. The value of SEM studies in elucidating the relationship between members of the genus Echinoparyphium in Britain/Europe and those in Africa, Asia and North America is suggested.

Intra-host competition between co-infecting digeneans within a bivalve second intermediate host: dominance by priority-effect or taking advantage of others?

We experimentally investigated the interactions between two parasites known to manipulate their host's phenotype, the trematodes Acanthoparyphium sp. and Curtuteria australis, which infect the cockle Austrovenus stutchburyi. The larval stages of both species encyst within the tissue of the bivalve's muscular foot, with a preference for the tip of the foot. As more individuals accumulate at that site, they impair the burrowing behaviour of cockles and increase the probability of the parasites' transmission to a bird definitive host. However, individuals at the foot tip are also vulnerable to non-host predators in the form of foot-cropping fish which selectively bite off the foot tip of exposed cockles. Parasites encysted at the foot base are safe from such predators although they do not contribute to altering host behaviour, but nevertheless benefit from host manipulation as all parasites within the cockle are transmitted if it is ingested by a bird. Experimental infection revealed that Acanthoparyphium sp. and C. australis have different encystment patterns within the host, with proportionally fewer Acanthoparyphium metacercariae encysting at the foot tip than C. australis. This indicates that Acanthoparyphium may benefit indirectly from C. australis and incur a lower risk of non-host predation. However, in co-infections, not only did C. australis have higher infectivity than Acanthoparyphium, it also severely affected the latter's infection success. The asymmetrical strategies and interactions between the two species suggest that the advantages obtained from exploiting the host manipulation efforts of another parasite might be offset by traits such as reduced competitiveness in co-infections.

Food makes you a target: disentangling genetic, physiological, and behavioral effects determining susceptibility to infection.

Genetics, physiology, and behavior are all expected to influence the susceptibility of hosts to parasites. Furthermore, interactions between genetic and other factors are suggested to contribute to the maintenance of genetic polymorphism in resistance when the relative susceptibility of host genotypes is context dependent. We used a maternal sibship design and long- and short-term food deprivation treatments to test the role of family-level genetic variation, body condition, physiological state, and foraging behavior on the susceptibility of Lymnaea stagnalis snails to infection by a trematode parasite that uses chemical cues to locate its hosts. In experimental exposures, we found that snails in the long-term food deprivation treatment contracted fewer parasites than snails that were continuously well-fed, possibly because well-fed snails grew larger and attracted more transmission stages. When we kept the long-term feeding rates the same, but manipulated the physiological state and foraging behavior of the snails with short-term food deprivation treatment, we found that snails that were fed before the exposure contracted more parasites than snails that were fed during the exposure. This suggests that direct physiological effects of food processing, but not foraging behavior, predisposed snails to infection. Feeding treatments also affected the family-level variation in snail susceptibility, suggesting that the relative susceptibility of host genotypes was context dependent.

Larval digenean community parasitizing the freshwater snail, Biomphalaria peregrina (Pulmonata: Planorbidae), from a temporary pond in Patagonia, Argentina.

Knowledge of population dynamics of parasites in freshwater snails from South America is scarce. The objective of the present study was to describe the infection dynamics of larval digeneans in the planorbid snail, Biomphalaria peregrina , during 2 sampling periods in a Patagonian temporary pond. In total, 1,003 snails were examined. Rediae of Notocotylus biomphalariae and Echinoparyphium sp., sporocysts of Cotylurus sp., and metacercariae of the 2 latter species were found. The overall prevalence was significantly higher in the second sampling period, always as single-species infections in the hepatopancreas. The presence of larvae in the first sampled snails of the second hydroperiod indicated that parasitized snails survive drought. Both species exhibited different seasonal prevalence patterns, with Echinoparyphium sp. present in all sampling months. Metacercariae of Echinoparyphium sp. occurred in the heart and kidney, and those of Cotylurus sp. between organs. No significant differences in overall prevalence of metacercariae were found, and a progressive rise in prevalence from spring to summer for both species was observed. Almost all size classes of B. peregrina were infected with metacercariae of both species, but rediae and sporocysts were present only in snails larger than 3.1 mm. The predictability of the hydroperiod year after year, the tolerance of B. peregrina to drought, and the survival of infected specimens allows the parasite community to show a similar pattern of infection over time. This is the first study in Argentina analyzing the infection dynamics of digeneans of a pulmonate snail from a temporary pond.