Balancing sample sizes in parasitology: A standardized experimental infection method using faecal parasite eggs and aquatic intermediate hosts.

Some parasites with complex life-cycles are able to manipulate the behaviour of their intermediate hosts in a way that increases their transmission to the next host. Gammarids infected by the tapeworm Cyathocephalus truncatus (Cestoda: Spathebothriidea) are known to be more predated by fish than uninfected ones, but potential behavioural manipulation by the parasite has never been investigated. In this study, we tested the hypothesis that C. truncatus is able to manipulate the behaviour of one of its intermediate hosts, Gammarus pulex (Crustacea: Amphipoda). To assess if any behavioural change was linked to other phenotypic alterations, we also measured the immunity of infected and uninfected individuals and investigated the pathogenic effects of the parasite. Infected gammarids were significantly less photophobic than uninfected ones, but no effect of infection on the level of immune defence was found. The results on survival, swimming activity and oxygen consumption suggest that the parasite also has various pathogenic effects. However, the alteration in host phototaxis was not correlated to some of these pathogenic effects. Therefore, we propose that the modification in host reaction to light is a behavioural manipulation, explaining the previously observed increase of gammarid predation rate.

The similarity in species composition between two communities generally decays as a function of increasing distance between them. Parasite communities in vertebrate definitive hosts follow this pattern but the respective relationship in intermediate invertebrate hosts of parasites with complex life cycles is unknown. In intermediate hosts, parasite communities are affected not only by the varying vagility of their definitive hosts (dispersing infective propagules) but also by the necessary coincidence of all their hosts in environmentally suitable localities. As intermediate hosts often hardly move they do not contribute to parasite dispersal. Hence, their parasite assemblages may decrease faster in similarity with increasing distance than those in highly mobile vertebrate definitive hosts. We use published field survey data to investigate distance decay of similarity in trematode communities from three prominent coastal molluscs of the Eastern North-Atlantic: the gastropods Littorina littorea and Hydrobia ulvae, and the bivalve Cerastoderma edule. We found that the similarity of trematode communities in all three hosts decayed with distance, independently of local sampling effort, and whether or not the parasites used the mollusc as first or second intermediate host in their life cycle. In H. ulvae, the halving distance (i.e. the distance that halves the similarity from its initial similarity at 1 km distance) for the trematode species using birds as definitive hosts was approximately two to three times larger than for species using fish. The initial similarities (estimated at 1 km distance) among trematode communities were relatively higher, whereas mean halving distances were lower, compared to published values for parasite communities in vertebrate hosts. We conclude that the vagility of definitive hosts accounts for a high similarity at the local scale, while the strong decay of similarity across regions is a consequence of the low probability that all necessary hosts and suitable environmental conditions coincide on a large scale.

Infection parameters of Posthodiplostomum cuticola, a digenean parasite with a complex life-cycle, were investigated in fish (the second intermediate host) from 6 floodplain water bodies over 2 years. A broad range of factors related to abiotic characteristics of localities, density of the first intermediate (planorbid snails) and definitive (wading birds) hosts and fish community structure were tested for their effects on P. cuticola infection in juvenile and adult fish. Characters of the littoral zone and flood duration were found to be important factors for the presence of the first intermediate and definitive hosts. Visitation time of definitive bird hosts was also related to adult fish host density. Localities with P. cuticola infected fish were visited by a higher number of bird species. Infection of P. cuticola in fish and similarities in infection among fish host assemblages were correlated with fish host density and fish species composition. Parasite infection in both adult and juvenile fishes was associated with the slope of the bank and the bottom type, in particular in juvenile fish assemblages with snail host density. We conclude that habitat characteristics, snail host density and fish community structure contribute significantly to P. cuticola infection in fish hosts.

BackgroundParamphistomosis caused by Calicophoron daubneyi and fasciolosis caused by Fasciola hepatica are common parasitic diseases of livestock animals. Transmission of the diseases depends on the presence of intermediate hosts, i.e. freshwater gastropods such as lymnaeids. We carried out a 2-year-long study of the dynamics of the snail population acting as the intermediate host for these parasites, considering the population structure in terms of size/age and infection status. In addition, we determined the kinetics of trematode egg excretion in grazing cows. Generalized Additive Models (GAMs) were used to analyze the associations between different response variables and snail size, sampling month and weather-related variables.ResultsOf the molluscan species examined, Galba truncatula, Radix peregra, Anisus (Anisus) leucostoma and Pisidium casertanum (n = 2802), only G. truncatula was infected with C. daubneyi or F. hepatica, at prevalence rates of 8.2% and 4.4% respectively. The probability of infection with C. daubneyi or F. hepatica was linearly related to snail size, although in different ways (negative for C. daubneyi and positive for F. hepatica). The total snail population increased in winter, when specimens of all size classes were found. Infected snails were more abundant during spring-autumn. Mature cercariae of both parasites were found in most seasons. In the statistical models, the sampling month accounted for a high percentage (71.9–78.2%) of the observed variability in snail abundance. The inclusion of climatic variables in the models moderately increased the percentage of deviance explained (77.7–91.9%). Excretion of C. daubneyi eggs in cow faeces was always higher than that of F. hepatica eggs.ConclusionsParticular care should be taken to prevent pastures and the surrounding environment being contaminated with parasite eggs during winter-spring, when the number of snails susceptible to miracidial infections is maximal. This is therefore the optimal time for treating grazing animals. Nevertheless, control of trematodosis based only on chemotherapy is difficult in an area such as the study area, where environmental factors favour the regular appearance of snail populations harbouring mature cercariae.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1892-8) contains supplementary material, which is available to authorized users.

Many populations normally experience high levels of mortality throughout larval development, but this is generally overlooked with laboratory experimental protocols. Evidence suggests that mortality is nonrandom in natural tadpole populations, so high survivorship, typical of laboratory populations, may poorly represent populations in nature. We compared survival, growth and development, and population variance of tadpoles in natural ponds with those in the laboratory at low and high densities. In the laboratory, high-density groups were reared with no selection and with selection imposed against different size classes to identify if, and how, mortality influences natural tadpole populations and to investigate whether imposing selection against certain size classes produces responses more consistent with those observed in natural systems. Our results suggest that selective mortality removes smaller individuals in natural populations. We demonstrate that introducing selection against small individuals artificially, in the laboratory, results in individual growth and development, population variance, and statistical power that more closely resembles that observed in natural populations. This is important from an ecological perspective because it demonstrates how selection acts on natural tadpole populations. More importantly, this demonstrates that laboratory experiments can be designed to provide better qualitative estimates for responses of natural populations by considering and simulating natural rates of mortality.

The nature of gene flow in parasites with complex life cycles is poorly understood, particularly when intermediate and definitive hosts have contrasting movement potential. We examined whether the fine-scale population genetic structure of the diphyllobothriidean cestode Schistocephalus solidus reflects the habits of intermediate threespine stickleback hosts or those of its definitive hosts, semi-aquatic piscivorous birds, to better understand complex host-parasite interactions. Seventeen lakes in the Cook Inlet region of south-central Alaska were sampled, including ten in the Matanuska-Susitna Valley, five on the Kenai Peninsula, and two in the Bristol Bay drainage. We analyzed sequence variation across a 759 bp region of the mitochondrial DNA (mtDNA) cytochrome oxidase I region for 1,026 S. solidus individuals sampled from 2009-2012. We also analyzed allelic variation at 8 microsatellite loci for 1,243 individuals. Analysis of mtDNA haplotype and microsatellite genotype variation recovered evidence of significant population genetic structure within S. solidus. Host, location, and year were factors in structuring observed genetic variation. Pairwise measures revealed significant differentiation among lakes, including a pattern of isolation-by-distance. Bayesian analysis identified three distinct genotypic clusters in the study region, little admixture within hosts and lakes, and a shift in genotype frequencies over time. Evidence of fine-scale population structure in S. solidus indicates that movement of its vagile, definitive avian hosts has less influence on gene flow than expected based solely on movement potential. Observed patterns of genetic variation may reflect genetic drift, behaviors of definitive hosts that constrain dispersal, life history of intermediate hosts, and adaptive specificity of S. solidus to intermediate host genotype.

In the presented article, the phenomenon of multiparasitism in the rock agama (Paralaudakia caucasia Eichw., 1831) by the malarial species Plasmodium caucasicaTelford, 2013, larvae of tissue nematodes from the Onchocercidae and Spirocercidae families were studied. The host-parasite relationships in animals infected by the aforementioned blood and tissue parasites, as well as the influence of these parasites on their hosts, were investigated. From 1996 to 2005, agamid lizards were captured in the semi-desert territories of Eastern Azerbaijan. Fifteen years later, the blood smears of these lizards were re-examined in light of the new findings. In one mature male rock agama, joint parasitism by plasmodia and larvae of tissue nematodes was recorded. Spirocerca lupi (Rud., 1819) (Nematoda: Spirurida) is endemic in many tropical and subtropical regions worldwide. The canine predators may be secondary hosts for these parasitic nematodes. In general, S. lupi has a complex life cycle with both intermediate and paratenic hosts. Meanwhile, S. lupi has an enigmatic peculiarity: infection of canids from intermediate hosts is extremely limited. Probably, coprophagous beetles are the intermediate hosts, ingesting eggs that are contaminated with larvae. The male rock agama, infected by the parasites mentioned above, was very exhausted and soon died. Probably, tissue nematodes have a generalized effect on the paratenic host. The presence of a parasite in the chest of an infected animal, similar in morphology to free-living flatworms, was exciting. The second parasite found in the rock agama is the hemosporidian species P. caucasica. The area inhabited by this parasite spans a large territory from the Caucasus to Pakistan. P. caucasica is a common parasite in rock agamas and can significantly weaken them in case of mass or joint infection with various invasive agents. We described the pathogenic effect of S. lupi larvae, characterized by a general weakening leading to mortality, as well as an increase in the intensity of infection of blood cells by plasmodia compared to lizards infected only with P. caucasica. A conclusion about the interaction between parasites and hosts in rock agamas can be drawn only after a comprehensive study.

Complex life cycles are a hallmark characteristic of many parasites; however, little is known about the process by which life cycles become more complex through the addition of hosts. Paratenic hosts are present in the life cycles of several phylogenetically distinct groups of helminths; this suggests that they may play a key role during this process. This study examined the development of metacercariae of Halipegus eccentricus within intermediate microcrustacean and odonate paratenic hosts. Then a comparative approach was used to evaluate how life history traits of H. eccentricus within the anuran definitive hosts differ between metacercariae of the same age that developed within an intermediate ostracod host or a paratenic odonate host. The results of this study indicate that metacercariae of H. eccentricus do not grow at the same rate in different intermediate hosts, and significant differences exist in growth within intermediate and paratenic hosts. Individuals from odonate paratenic hosts always had larger bodies and suckers than those of metacercariae of the same age that develop within microcrustacean intermediate hosts. Furthermore, metacercariae from odonates were more successful in establishing and migrating in definitive anuran hosts. Last, individuals from paratenic hosts began reproducing earlier within anuran definitive hosts than age-matched worms that develop within the intermediate hosts. Collectively these results suggest that the variation in body and sucker sizes within odonate and microcrustacean hosts may carry over to the definitive host and in the case of H. eccentricus using the paratenic host increases transmission and alters other life history traits within definitive hosts. These results indicate that using a paratenic host can affect the success of parasites in subsequent hosts, and therefore these hosts may provide benefits other than just increasing transmission by bridging an ecological gap.

How complex life cycles of parasites are maintained is still a fascinating and unresolved topic. Complex life cycles using three host species, free-living stages, asexual and sexual reproduction are widespread in parasitic helminths. For such life cycles, we propose here that maintaining a second intermediate host in the life cycle can be advantageous for the individual parasite to increase the intermixture of different clones and therefore decrease the risk of matings between genetically identical individuals in the definitive host. Using microsatellite markers, we show that clone mixing occurs from the first to the second intermediate host in natural populations of the eye-fluke Diplostomum pseudospathaceum. Most individuals released by the first intermediate host belonged to one clone. In contrast, the second intermediate host was infected with a diverse array of mostly unique parasite genotypes. The proposed advantage of increased parasite clone intermixture may be a novel selection pressure favouring the maintenance of complex life cycles.

The cestode parasite Schistocephalus solidus' growth is limited by the size of its second intermediate host, the three-spined stickleback, Gasterosteus aculeatus. S. solidus should thus prefer a large stickleback as host. Since the stickleback is a predator of the parasite's previous intermediate host, a small copepod, the stickleback that consumes the infected copepod will probably be of a size for which this copepod has the optimal prey size. The optimal foraging decision of the stickleback may or may not be compatible with the parasite's preference. Infected copepods are present in early summer when both many size classes of young of the year and adult sticklebacks are potential predators. We offered laboratory bred three-spined sticklebacks of four size classes individually the choice among five prey types: two size classes of copepods, two classes of Daphnia of corresponding size as alternative prey and a third Daphnia size class that was larger than the larger copepod. We found that small copepods, the potential hosts of S. solidus, were most accepted by the smallest sticklebacks of about 1.5 cm of length, larger fish consumed a decreasing proportion; fish larger than 3.8 cm did not consume them at all. Experience with copepods over several weeks increased the acceptance for this prey to some extend but hardly in the largest fish. This suggests that S. solidus will end up usually in sticklebacks that are too small for the parasite so that it has to allow its host's further growth after infection to reach its definitive size.

A chromosome-level genome of the human blood fluke Schistosoma japonicum identifies the genomic basis of host-switching.

A manipulative parasite increasing an antipredator response decreases its vulnerability to a nonhost predator

Transmission dynamics of Toxoplasma gondii along an urban–rural gradient

Many parasites with complex life cycles are known to modify their host phenotype to enhance transmission from the intermediate host to the definitive host. Several earlier studies explored these effects in acanthocephalan and trematode parasites, especially in aquatic ecosystems; however, much less is known about parasite-mediated alterations of host behavior in terrestrial systems involving nematodes. Here, we address this gap by investigating a trophically transmitted nematode (Pterygodermatites peromysci) that uses a camel cricket (Ceuthophilus pallidipes) as the intermediate host before transmission to the final host, the white-footed mouse (Peromyscus leucopus). In a laboratory experiment, we quantified the anti-predatory responses of the cricket intermediate host using simulated predator cues. Results showed a decrease in jumping performance among infected crickets as compared with uninfected crickets, specifically in terms of frequency of jumps and jumping distance. Additionally, the relationship between parasite load and frequency of jumps is negatively correlated with the intensity of infection. These behavioral modifications are likely to increase vulnerability to predation by the definitive host. An analysis of the age-intensity pattern of infection in natural cricket populations appears to support this hypothesis: parasites accumulate with age, peak at an intermediate age class before the intensity of infection decreases in older age groups. We suggest that older, heavily infected crickets are preferentially removed from the population by predators because of increased vulnerability. These results show that cricket intermediate hosts infected with P. peromysci have diminished jumping performance, which is likely to impair their anti-predatory behavior and potentially facilitate parasite transmission.

Both theoretical arguments and empirical evidence suggest that parasite transmission depends on host density. In helminths with complex life cycles, however, it is not clear which host, if any, is the most important. Here, the relationships between the abundance of metacercariae in second intermediate hosts, and the local density of both the first and second intermediate hosts of two trematode species, are investigated. Samples of the snail Potamopyrgus antipodarum, the amphipod Paracalliope fluviatilis and the isopod Austridotea annectens were collected from ten stations in a New Zealand lake. In the trematode Coitocaecum parvum, neither the density of the snail first intermediate host nor that of the amphipod second intermediate host correlated with infection levels in amphipods. In contrast, in the trematode Microphallus, infection levels in isopod second intermediate hosts were positively associated with isopod density and negatively associated with the density of snail first intermediate hosts. These relationships are explained by a negative correlation between snail and isopod densities, mediated in part by their different use of macrophyte beds in the lake. Overall, the results suggest that, at least for Microphallus, local infection levels depend on local intermediate host densities.