Is the local environment more important than within-host interactions in determining coinfection?

Host populations often vary in the magnitude of coinfection they experience across environmental gradients. Furthermore, coinfection often occurs sequentially, with a second parasite infecting the host after the first has established a primary infection. Because the local environment and interactions between coinfecting parasites can both drive patterns of coinfection, it is important to disentangle the relative contributions of environmental factors and within-host interactions to patterns of coinfection. Here, we develop a conceptual framework and present an empirical case study to disentangle these facets of coinfection. Across multiple lakes, we surveyed populations of five damselfly (host) species and quantified primary parasitism by aquatic, ectoparasitic water mites and secondary parasitism by terrestrial, endoparasitic gregarines. We first asked if coinfection is predicted by abiotic and biotic factors within the local environment, finding that the probability of coinfection decreased for all host species as pH increased. We then asked if primary infection by aquatic water mites mediated the relationship between pH and secondary infection by terrestrial gregarines. Contrary to our expectations, we found no evidence for a water mite-mediated relationship between pH and gregarines. Instead, the intensity of gregarine infection correlated solely with the local environment, with the magnitude and direction of these relationships varying among environmental predictors. Our findings emphasize the role of the local environment in shaping infection dynamics that set the stage for coinfection. Although we did not detect within-host interactions, the approach herein can be applied to other systems to elucidate the nature of interactions between hosts and coinfecting parasites within complex ecological communities.

Male-Male Competition Causes Parasite-Mediated Sexual Selection for Local Adaptation.

AbstractSexual selection has been suggested to accelerate local adaptation and promote evolutionary rescue through several ecological and genetic mechanisms. Condition-dependent sexual selection has mainly been studied in laboratory settings, while data from natural populations are lacking. One ecological factor that can cause condition-dependent sexual selection is parasitism. Here, we quantified ectoparasite load (Arrenurus water mites) in a natural population of the common bluetail damselfly (Ischnura elegans) over 15 years. We quantified the strength of sexual selection against parasite load in both sexes and experimentally investigated the mechanisms behind such selection. Then we investigated how parasite resistance and tolerance changed over time to understand how they might influence population density. Parasites reduced mating success in both sexes, and sexual selection was stronger in males than in females. Experiments show that male-male competition is a strong force causing precopulatory sexual selection against parasite load. Although parasite resistance and male parasite tolerance increased over time, suggestive of increasing local adaptation against parasites, no signal of evolutionary rescue could be found. We suggest that condition-dependent sexual selection facilitates local adaptation against parasites and discuss its effects in evolutionary rescue.

Studies on the life cycle of Pleurogenoides wayanadensis Shinad & Prasadan, 2018 (Digenea: Pleurogenidae) from the Western Ghats, India.

The life cycle of Pleurogenoides wayanadensis Shinad & Prasadan, 2018, infecting the frogs Hoplobatrachus tigerinus and Euphlyctis cyanophlyctis, is elucidated in this study. All the life cycle stages from egg to egg-producing adults were elucidated under natural conditions and successfully established in the laboratory. The life cycle took about 58 to 65 days for completion. Miracidia were released by teasing the eggs with fine needles. Sporocysts were found in the freshwater snail, Bithynia (Digoniostoma) pulchella, collected from paddy fields at Payode, Western Ghats, Wayanad region, in the months of October and November 2019. Cercariae were of the virgulate xiphidiocercous type. Metacercariae were recovered from the eyes of the damselfly naiads of the species Ischnura sp. and Copera sp., and the thorax and abdomen of the dragonfly naiads, Orthetrum sp. The metacercariae showed progenetic development. The growth and development of the metacercariae in the naiads that were exposed to cercariae, and development of the trematode in frogs that were force-fed with encysted metacercariae, have been studied at regular intervals. The prepatent period is 14-19 days. The present life cycle study of a Pleurogenoides spp. forms the seventh report from the world, fourth report from India and the third from Kerala.

Parasitic gut infection in Libellula pulchella causes functional and molecular resemblance of dragonfly flight muscle to skeletal muscle of obese vertebrates.

We previously demonstrated the existence of a naturally occurring metabolic disease phenotype in Libellula pulchella dragonflies that shows high similarity to vertebrate obesity and type II diabetes, and is caused by a protozoan gut parasite. To further mechanistic understanding of how this metabolic disease phenotype affects fitness of male L. pulchella in vivo, we examined infection effects on in situ muscle performance and molecular traits relevant to dragonfly flight performance in nature. Importantly, these traits were previously shown to be affected in obese vertebrates. Similarly to obesity effects in rat skeletal muscle, dragonfly gut infection caused a disruption of relationships between body mass, flight muscle power output and alternative pre-mRNA splicing of troponin T, which affects muscle calcium sensitivity and performance in insects and vertebrates. In addition, when simulated in situ to contract at cycle frequencies ranging from 20 to 45 Hz, flight muscles of infected individuals displayed a left shift in power-cycle frequency curves, indicating a significant reduction in their optimal cycle frequency. Interestingly, these power-cycle curves were similar to those produced by flight muscles of non-infected teneral (i.e. physiologically immature) adult L. pulchella males. Overall, our results indicate that the effects of metabolic disease on skeletal muscle physiology in natural insect systems are similar to those observed in vertebrates maintained in laboratory settings. More generally, they indicate that study of natural, host-parasite interactions can contribute important insight into how environmental factors other than diet and exercise may contribute to the development of metabolic disease phenotypes.

Negative covariance between water mite and gregarine parasitism for adult dragonflies, Leucorrhinia intacta (Hagen): an age-related pattern?

Studies on parasite-mediated selection often focus on single parasite taxa infecting single species of hosts. However, host populations experience infections by multiple parasite taxa simultaneously; coinfection is expected to influence how host- and/or parasite-related factors affect host exposure and susceptibility to various parasites, and the resulting patterns of infection. We sampled adult dragonflies from a population of Leucorrhinia intacta (Hagen) in eastern Ontario, Canada. Dragonflies were exposed to parasitism by both water mites (Arrenuridae) and gregarines (Eugregarinidae). We tested for covariation between these ecto- and endoparasites, while considering potential sex and age biases in host sampling and patterns of infection. Mite parasitism differed dramatically between host sexes: nearly all collected males were parasitized, whereas only half of females were infested. This was likely due to differences in age distributions between sexes in sampled dragonflies. Water mite and gregarine parasitism showed strong, negative covariation, and coinfection occurred far less often than expected by chance, although these patterns were restricted to samples of females which, unlike male samples, likely included many old and young dragonflies. We report the first observation of negative covariation between internal and external parasite taxa in an anisopteran host and suggest this relationship between water mites and gregarines may be more widespread among Odonata and perhaps other insects than previously surmised. We advance hypotheses based on host age-parasitism relationships as well as variable parasite-mediated selection to help explain the sex specificity of observed coinfection patterns in our samples.

Intra- and intersexual differences in parasite resistance and female fitness tolerance in a polymorphic insect.

To understand host-parasite interactions, it is necessary to quantify variation and covariation in defence traits. We quantified parasite resistance and fitness tolerance of a polymorphic damselfly (Ischnura elegans), an insect with three discrete female colour morphs but with monomorphic males. We quantified sex and morph differences in parasite resistance (prevalence and intensity of water mite infections) and morph-specific fitness tolerance in the females in natural populations for over a decade. There was no evidence for higher parasite susceptibility in males as a cost of sexual selection, whereas differences in defence mechanisms between female morphs are consistent with correlational selection operating on combinations of parasite resistance and tolerance. We suggest that tolerance differences between female morphs interact with frequency-dependent sexual conflict, which maintains the polymorphism locally. Host-parasite interactions can therefore shape intra- and intersexual phenotypic divergence and interfere with sexual selection and sexual conflict.

Behavioural modification of personality traits: testing the effect of a trematode on nymphs of the red damselfly Xanthocnemis zealandica.

Research on animal personality is increasingly demonstrating that individuals in a population are characterised by distinct sets of behavioural traits that show consistency over time and across different situations. Parasites are known to alter the behaviour of their hosts, although their role in shaping host personality remains little studied. Here, we test the effect of trematode infection on two traits of their host's personality, activity and boldness, in nymphs of the red damselfly Xanthocnemis zealandica. Genetic analyses indicate that the undescribed trematode species falls within the superfamily Microphalloidea. Results of laboratory behavioural tests indicate that the two behavioural traits are related to each other: bolder individuals also show higher levels of spontaneous activity than shy ones. However, parasite infection had no effect on either of these behaviours or on their repeatability over three separate testing sessions. Although our findings suggest that this trematode does not influence personality traits of the damselfly host, it remains possible that other standard personality traits not tested here (exploratory tendency, aggressiveness) are affected by infection.

Resistance to Arrenurus spp. Parasitism in Odonates: Patterns Across Species and Comparisons Between a Resistant and Susceptible Host.

Some adult odonates resist parasitism by larval water mites (Arrenurus spp.) with melanotic encapsulation, in which the mite's stylestome is clogged and the mite starves. In summer 2014, we counted the engorged and resisted mites on 2,729 adult odonates sampled by aerial net at 11 water bodies in Greenville Co. and Pickens Co., SC, and tested the hypothesis that the frequency and intensity of resistance correlates with parasite prevalence (the percentage of parasitized hosts). Resistance prevalence (the percentage of parasitized hosts that resisted at least one mite) varied significantly among host species, exceeding 60% for Argia fumipennis(Burmeister) and Celithemis fasciata Kirby but less than 20% for other species. However, neither resistance prevalence nor mean resistance intensity (mean percentage of resisted mites on resisting hosts) correlated with parasite prevalence. We described potential effects of parasitism on host development ofA. fumipennis and Pachydiplax longipennis(Burmeister) by comparing the percent asymmetry of forewing lengths between parasitized and unparasitized individuals. There was no significant difference in asymmetry for either males or females of A. fumipennis, or males of Pa. longipennis(females were not sampled). We also evaluated differences in melanotic encapsulation between A. fumipennis, which readily encapsulates mites in nature, and Pa. longipennis We inserted a 2.0-mm piece of sterile monofilament line into the thorax of captured individuals for 24 h and compared mean gray value scores of inserted and emergent ends using Image-J software. There was no difference in melanotic encapsulation between species.

It's a predator-eat-parasite world: how characteristics of predator, parasite and environment affect consumption.

Understanding the effects of predation on disease dynamics is increasingly important in light of the role ecological communities can play in host-parasite interactions. Surprisingly, however, few studies have characterized direct predation of parasites. Here we used an experimental approach to show that consumption of free-living parasite stages is highly context dependent, with significant influences of parasite size, predator size and foraging mode, as well as environmental condition. Among the four species of larval trematodes and two types of predators (fish and larval damselflies) studied here, parasites with larger infective stages (size >1,000 µm) were most vulnerable to predation by fish, while small-bodied fish and damselflies (size <10 mm) consumed the most infectious stages. Small parasite species (size approx. 500 µm) were less frequently consumed by both fish and larval damselflies. However, these results depended strongly on light availability; trials conducted in the dark led to significantly fewer parasites consumed overall, especially those with a size of <1,000 µm, emphasizing the importance of circadian shedding times of parasite free-living stages for predation risk. Intriguingly, active predation functioned to help limit fishes' infection by directly penetrating parasite species. Our results are consistent with established theory developed for predation on zooplankton that emphasizes the roles of body size, visibility and predation modes and further suggest that consumer-resource theory may provide a predictive framework for when predators should significantly influence parasite transmission. These results contribute to our understanding of transmission in natural systems, the role of predator-parasite links in food webs and the evolution of parasite morphology and behavior.

MORPHOLOGY AND ULTRASTRUCTURE OF TWO SCHISTOTAENIID CYSTICERCOIDS (CESTODA: CYCLOPHYLLIDEA) FROM THE HAEMOCOELE OF THE DRAGONFLY LARVAE.

Two cysticercoids, belonging to ascocercus type, namely euascocercus and multicercus, were found in haemocoele of dragonfly larvae of the genus Aeshna from the lakes of the Magadan Province. The cysticercoid of Schistotaenia srivastavai Raush, 1970 (euascocercus) is formed of the outer (exocyst) and inner (endocyst) envelopes, containing the scolex and larval strobila. The outer and inner surfaces of the exocyst are represented by the tegument covered with microvilli. The microvilli of the outer tegument are restricted by the surface layer, consisting of granular and fibrillar material, and possess different structures at different stages of post-embryonic development. The multicercus of Mircia shigini (Konyaev et Gulyaev, 2006) is able to multiply asexually by the endogenous budding. The daughters' individuals are formed in the envelope of the multicercus that represents the tegument bearing microvilli. These microvilli are also restricted by the surface layer. The morphology and development of each individual cysticercoid of the multicercus is similar to those of euascocercus. The production of a great amount of cysticercoids, and the presence of the surface layer resembling the laminated layer of Echinococcus, relates multicercus to hydatid cysts.

The life history of Pleurogenoides malampuzhensis sp. nov. (Digenea: Pleurogenidae) from amphibious and aquatic hosts in Kerala, India.

The life-cycle stages of Pleurogenoides malampuzhensis sp. nov. infecting the Indian bullfrog Hoplobatrachus tigerinus (Daudin) and the skipper frog Euphlyctis cyanophlyctis (Schneider) occurring in irrigation canals and paddy fields in Malampuzha, which forms part of the district of Palakkad, Kerala, are described. The species is described, its systematic position discussed and compared with the related species, P. gastroporus (Luhe, 1901) and P. orientalis (Srivastava, 1934). The life-cycle stages, from cercaria to egg-producing adult, were successfully established in the laboratory. Virgulate xiphidiocercariae emerged from the snail Digoniostoma pulchella (Benson). Metacercariae are found in muscle tissues of dragonfly nymphs and become infective to the frogs within 22 days. The pre-patent period is 20 days. Growth and development of both metacercariae and adults are described.

Ultrastructural study of protective envelopes in Dioecocestus asper (Cestoda: Dioecocestidae) megalocercus.

The megalocercus of Dioecocestus asper (Mehlis 1831) from the haemocoele of dragonfly larvae possesses two envelopes: outer (exocyst) and inner (endocyst) ones. The exocyst contains the large endocyst and larval strobila with scolex attached to the latter. Outer and inner surfaces of these envelopes are organized as the tegument and have some structural differences. The exocyst is covered with slender microvilli. Its outer tegument contains numerous mitochondria; the inner one is filled with lipid droplets released into the exocyst's cavity. The well-developed protonephridial (excretory) system consisting of flame cells, collecting ducts and canals is the unique feature of the exocyst, noted for the first time. Thick (more, then 50 microm) distal cytoplasm of the outer tegument of the endocyst is the place of accumulation of uniform globules looking like a hyaloid layer. This outer layer together with underlying fibrous layer (up to 20 microm), apparently, protect the scolex and larval strobila during the transfer through feather clump in the stomach of grebes, definitive hosts of D. asper. Muscle cells of both envelopes retain their synthetic activity even in the fully developed metacestode. Probably, they are the main structural element, which produces fibers of the extracellular matrix and maintains the integrity of protective envelopes of the megalocercus.

Association between host's genetic diversity and parasite burden in damselflies.

Recent research indicates that low genetic variation in individuals can increase susceptibility to parasite infection, yet evidence from natural invertebrate populations remains scarce. Here, we studied the relationship between genetic heterozygosity, measured as AFLP-based inbreeding coefficient fAFLP , and gregarine parasite burden from eleven damselfly, Calopteryx splendens, populations. We found that in the studied populations, 5-92% of males were parasitized by endoparasitic gregarines (Apicomplexa: Actinocephalidae). Number of parasites ranged from none to 47 parasites per male, and parasites were highly aggregated in a few hosts. Mean individual fAFLP did not differ between populations. Moreover, we found a positive association between individual's inbreeding coefficient and parasite burden. In other words, the more homozygous the individual, the more parasites it harbours. Thus, parasites are likely to pose strong selection pressure against inbreeding and homozygosity. Our results support the heterozygosity-fitness correlation hypothesis, which suggests the importance of heterozygosity for an individual's pathogen resistance.

Maintenance of polymorphic females: do parasites play a role?

The role of parasites in explaining maintenance of polymorphism is an unexplored research avenue. In odonates, female-limited color polymorphism (one female morph mimicking the conspecific male and one or more gynochromatic morphs) is widespread. Here we investigated whether parasitism contributes to color polymorphism maintenance by studying six species of female dimorphic damselflies using large databases of field-collected animals. We predicted that androchrome females (male mimics) would be more intensively parasitized than gynochrome females which is, according to previous studies, counterbalanced by the advantages of the former when evading male harassment compared to gynochrome females. Here we show that in Ischnura denticollis and Enallagma novahispaniae, androchrome females suffer from a higher degree of parasitism than gynochromatic females, and contrary to prediction, than males. Thus, our study has detected a correlation between color polymorphism and parasitic burden in odonates. This leads us to hypothesize that natural selection, via parasite pressure, can explain in part how androchrome and gynochrome female color morphs can be maintained. Both morphs may cope with parasites in a different way: given that androchrome females are more heavily parasitized, they may pay a higher fecundity costs, in comparison to gynochrome females.

Relative geographic range of sibling species of host damselflies does not reliably predict differential parasitism by water mites.

BACKGROUND: One of the main challenges in evolutionary parasitology is to determine the factors that explain variation among host species in parasitism. In this study, we addressed whether host phylogeny or ecology was important in determining host species use by water mites. Parasitism (prevalence and intensity) by Arrenurus water mites was examined in relation to geographic distribution of host damselflies from sibling species pairs. In addition, the likelihood of putative mite species parasitizing both species of a host species pair was explored. RESULTS: A total of 1162 damselflies were examined for water mites across four sites in Southeastern Ontario. These damselflies represent ten species (five closely related host species pairs) in the Coenagrionidae. Only two of the five species pairs showed near significant or significant differences in prevalence of infection by mites. In one of those species comparisons, it was the less widespread host that had higher water mite prevalence and in the other species comparison, the less widespread host species had lower water mite prevalence. Only one of the five pairs showed a significant difference in intensity of infection; intensity was higher in the species with a smaller geographic distribution. Based on the COI barcode, there were nine water mite clades (OTU) infecting these ten host species. Three Arrenurus OTUs may be host monospecific, four OTUs were specific to a given host species pair, and two OTUs infected at least three host species. Host species in each species pairs tend to share at least one of the Arrenurus OTU. No striking differences in mite species diversity were found among species in any species pair. Finally, the Arrenurus examined in this study appear to be ecological specialists, restricted to a particular type of habitat, parasitizing few to many of the host species present in that site or habitat. CONCLUSIONS: Although differences in levels of parasitism by water mites exist for some closely related hosts species, no such differences were found between other related host species. Differences in geographic range of related host species does not reliably explain differential levels of parasitism by water mites.

[On the life cycle and morphology of metacestodes Dioecocestus asper (Cyclophyllidea: Dioecocestidae)].

Metacestodes Dioecocestus asper (Cyclophyllidea) habe been found in larvae of dragonflies Aeshna spp. (Odonata, Anisoptera) from the lakes of the Upper Kolyma and northern Okhotsk seaboard basins (3.8 +/- 0.46 % of n = 1730 and 0.09 +/- 0.09 % of n = 1065, respectively) for the first time. Thus, participation of dragonflies in lifecycle of Dioecocestidae has been proved. Larvagenesis of D. asper (from spontaneously infested hosts) have been retraced from the stage of primary lacuna up to fully-developed metacestode. Process of development of the metacestode D. asper includes two invaginations. The first one follows the start of exogenous formation of undifferentiated anlage of the cystoscolex; as the result, the latter settles to the bottom of the forming two-layered exocyst homologous to the cercomer. Subsequent development of the prospecive part and the endocyst occurs in the closed amniotic cavity of the sacciform exocyst, the back pole of which preserves embryonic hooks and the front pole possesses the invaginational channel. At the stage of early scolexogenesis, the endocyst with the anlage of the strobila and the scolex separates from the exocyst and the excretory atrium forms on the back pole of endocyst. At the stage of late scolexogenesis, the formation of the larval strobila (differentiation of the sex) is completed and the second invagination starts. Strong retractors pull bottom of the stribila into the endocyst; simultaneously, the front part of it is also turned inside. The invaginational pore of the endocyst of fully-developed metacestode D. asper remains open; the scolex with the most part of the strobila is located outside the endocyst in the cavity of the exocyst. The encystment of the metacestode occurs when the temperature rises up to 40 degrees C (i. e., reaching the temperature of the definitive host). Morphogenesis of the uniquely large (among the cyclophyllid metacestodes developing in the invertebrate intermediate host) metacestode D. asper lasts for about two--three years and is associated with the developmental terms of the intermediate host, the larva of the Aeshna dragonfly, in climatic conditions of the Far North-East of Asia. Early stages of morphogenesis of D. asper were noted at the dragonfly larvae of the age of 0+ (at September) and 1+ (at June), and fully-developed metacestodes, only at elder ages. Owing to the extremely large size of D. asper metacestodes we refer them as a specific modification of cysticercoids: a megalocercus (Megalocercus). Common features were noted in the structure and morphogenesis of D. asper megalocercus and ascocerci of Schistotaeniidae. High morphological similarity of these metacestodes and their development in relative taxonomic groups of intermediate hosts, larvae of amphibiotic insects (the dragonfly larvae, and occasionally the larvae of mayflies or water-bugs), prove their belonging to the same philogenetically related group of metacestodes Cyclophyllidea, emerging independently from other groups of cysticercoids. We suggest naming this morpho-ecological group of larvocysts as ascocerci (or ascocysticercoids). The use of the name "ascocercus" as nominal nomenclature for group of larvocysts supposes the change of the name "ascocercus Schistotaeniidae" to "euascocercus".

Parasite susceptibility in an amphibian host is modified by salinization and predators.

Secondary salinization represents a global threat to freshwater ecosystems. Salts, such as NaCl, can be toxic to freshwater organisms and may also modify the outcome of species interactions (e.g. host-parasite interactions). In nature, hosts and their parasites are embedded in complex communities where they face anthropogenic and biotic (i.e. predators) stressors that influence host-parasite interactions. As human populations grow, considering how anthropogenic and natural stressors interact to shape host-parasite interactions will become increasingly important. We conducted two experiments investigating: (1) the effects of NaCl on tadpole susceptibility to trematodes and (2) whether density- and trait-mediated effects of a parasite-predator (i.e. damselfly) and a host-predator (i.e. dragonfly), respectively, modify the effects of NaCl on susceptibility to trematode infection. In the first experiment, we exposed tadpoles to three concentrations of NaCl and measured parasite infection in tadpoles. In the second experiment, we conducted a 2 (tadpoles exposed to 0 g L-1 NaCl vs. 1 g L-1 NaCl) x 4 (no predator, free-ranging parasite-predator (damselfly), non-lethal host-predator (dragonfly kairomone), and free-ranging parasite-predator + dragonfly kairomone) factorial experiment. In the absence of predators, exposure to NaCl increased parasite infection. Of the predator treatments, NaCl only caused an increase in parasite infection in the presence of the parasite-predator. However, direct consumption of trematodes caused a reduction in overall infection in the parasite-predator treatment. In the dragonfly kairomone treatment, a reduction in tadpole movement (i.e. trematode avoidance behavior) led to an increase in overall infection. In the parasite-predator + dragonfly kairomone treatment, antagonistic effects of the parasite-predator (reduction in trematode abundance) and dragonfly kairomone (reduction in parasite avoidance behavior) resulted in intermediate parasite infection. Collectively, these findings demonstrate that NaCl can increase amphibian susceptibility to parasites, and underscores the importance of considering predator-mediated interactions in understanding how contaminants influence host-parasite interactions.

Wrecking the Curve: Altered Functional Response of Tetragoneuria (Odonata: Corduliidae) Naiads Infected with Metacercariae of Haematoloechus floedae.

The ubiquity of host-parasite interactions and their potential for substantial representation, in terms of overall biomass, within ecosystems suggests that parasites have the capacity to influence energy flow within an ecosystem. Although the influence of certain parasites on prey behavior has been well documented, parasites could also exert an influence on ecosystem dynamics by influencing predator feeding behavior. The functional response of Tetragoneuria naiads was characterized by presenting naiads with varying abundances of Daphnia magna , after which a subset of the naiads were exposed to cercariae of Haematoloechus floedae, and the feeding trials repeated for both the control and exposed odonates. A type II functional response was chosen as an appropriate model for comparison. An indicator variable approach to nonlinear regression of the functional response data indicated that infected odonate naiads spent significantly more time foraging than they did before infection, whereas there was no significant change in the functional response of the control naiads. Infected odonates also had a slower rate of growth. These results imply a metabolic cost to infection of Tetragoneuria naiads by H. floedae that might be associated with the encapsulating response to the metacercariae that was observed in infected naiads.

Larval stages of Neoplagioporus elongatus (Goto and Ozaki, 1930) (Opecoelidae: Plagioporinae), with notes on potential second intermediate hosts.

The morphology of sporocysts and cercariae of Neoplagioporus elongatus (Goto and Ozaki, 1930) is described for the first time. A cotylomicrocercous cercaria obtained from the sorbeoconch snail Semisulcospira nakasekoae was confirmed to be the cercaria of N. elongatus, based on the degree of sequence identity of the COI gene to that of adult worms. Freshwater annelids (oligochaetes and leeches) and some aquatic insects (odonates) were demonstrated experimentally to be potential second intermediate hosts.

Evaluating the Biological and Ecological Factors Influencing Transmission of Larval Digenetic Trematodes: A Test of Second Intermediate Host Specificity of Two North American Halipegus Species.

Host specificity of parasites is a basic principle in parasitology; however, it is not easily measured. Previously, host specificity was calculated as the number of species that a parasite infected, but this is not an accurate description of host usage because some species are capable of being infected but do not contribute to the completion of the life cycle. Instead, measures of host specificity should take into consideration interactions between a parasite and a potential host species as well as interactions between current and subsequent hosts in the life cycle. The objectives of this study were to track the development of 2 trematode species, Halipegus eccentricus and Halipegus occidualis , in 3 phylogenetically and ecologically distinct microcrustacean second intermediate hosts, and then evaluate the extent to which each of these hosts contributed to transmission of each Halipegus species to the next odonate host in the life cycle. All 3 microcrustacean species exposed became infected with both species of Halipegus. The patterns of growth of H. eccentricus and H. occidualis were similar, but there were consistent differences in the rates of growth among the microcrustacean species in both Halipegus species. Regardless of host species infected, all individuals of both species were considered to be developmentally infective to the next host in the life cycle by 19 days postexposure (DPE) when they lost their excretory bladder. Worms of varying sizes were capable of surviving without this structure, suggesting that there is not a strong relationship between the rate of growth of the metacercariae and the development of their osmoregulatory system. Although Halipegus species were capable of living without an excretory bladder at 19 DPE, there were differences in their size and rates at which the 3 microcrustaceans contributed to transmission of the parasites to subsequent odonate hosts. Collectively, under controlled laboratory conditions, there was an approximate 2-fold difference in the average percentage of worms that established in odonates from the ostracod, Cypridopsis sp., than from the harpacticoid copepod, Phyllognathopus sp., and the difference was nearly 3-fold between Cypridopsis sp. and the cyclopoid copepod, Thermocyclops sp. Therefore, despite all 3 microcrustacean species becoming infected, not all species were equally suited for transmission and completion of the life cycle. Differences among the 3 microcrustacean species in cercaria ingestion, metacercarial growth and development, and odonate predation rates on infected microcrustacean species were important factors in determining transmission of the 2 Halipegus species to odonate hosts.