Parasite Infrapopulations Research Articles

Extensive transmission and variation in a functional receptor for praziquantel resistance in endemic Schistosoma mansoni.

Mass-drug administration (MDA) of human populations using praziquantel monotherapy has become the primary strategy for controlling and potentially eliminating the major neglected tropical disease schistosomiasis. To understand how long-term MDA impacts schistosome populations, we analysed whole-genome sequence data of 570 Schistosoma mansoni samples (and the closely related outgroup species, S. rodhaini) from eight countries incorporating both publicly-available sequence data and new parasite material. This revealed broad-scale genetic structure across countries but with extensive transmission over hundreds of kilometres. We characterised variation across the transient receptor potential melastatin ion channel, TRPMPZQ, a target of praziquantel, which has recently been found to influence praziquantel susceptibility. Functional profiling of TRPMPZQ variants found in endemic populations identified four mutations that reduced channel sensitivity to praziquantel, indicating standing variation for resistance. Analysis of parasite infrapopulations sampled from individuals pre- and post-treatment identified instances of treatment failure, further indicative of potential praziquantel resistance. As schistosomiasis is targeted for elimination as a public health problem by 2030 in all currently endemic countries, and even interruption of transmission in selected African regions, we provide an in-depth genomic characterisation of endemic populations and an approach to identify emerging praziquantel resistance alleles.

A molecular and epidemiological study of Grillotia (Cestoda: Trypanorhyncha) larval infection in Etmopterus spinax (Elasmobranchii: Squaliformes) in the Mediterranean Sea and Northeast Atlantic Ocean

Amongst other factors, topographic features can influence the genetic variability among populations of marine organisms. This applies to host species but also to their parasites, which are poorly studied regarding this aspect, as well as with regard to their use as bioindicators. In the present work, the ribosomal DNA (28S rDNA) was used to assess genetic diversity of Grillotia (Cestoda, Trypanorhyncha) larvae in one of its paratenic hosts, namely Etmopterus spinax, across five different regions (off Scotland, Celtic, Alboran and Balearic Seas and off Cyprus) belonging to three major geographic areas (Northeast Atlantic, western and eastern Mediterranean). The obtained sequences revealed a total of 18 polymorphic sites and 17 haplotypes, as well as significant values of variance throughout the five different regions. Reconstructed phylogenetic trees highlighted that all Grillotia sp. sequences formed a monophyletic group, but divergent lineages split into different main clades which were in relation to the area of origin, with a consistent cluster of sequences from the Atlantic Ocean, as well as another from the Eastern Mediterranean. In contrast, low genetic differentiation was observed between samples from Balearic and Alboran Seas, and with respect to Grillotia sp. larvae from the Gulf of Naples analysed in a previous study. Geographical differences in parasite infection descriptors (prevalence, abundance, and intensity) were assessed, revealing significant differences among the sampled regions.The present study indicates that geographical distance and submarine barriers affect not only the connectivity of hosts but also their parasite infrapopulations by limiting interpopulation dispersal. It underlines the usefulness of parasites as biological tags for the study of susceptible and data-poor host species such as deep-water sharks and its potential implications for host population management and protection measures.

Host body size, not host population size, predicts genome-wide effective population size of parasites.

The effective population size (Ne) of an organism is expected to be generally proportional to the total number of individuals in a population. In parasites, we might expect the effective population size to be proportional to host population size and host body size, because both are expected to increase the number of parasite individuals. However, among other factors, parasite populations are sometimes so extremely subdivided that high levels of inbreeding may distort these predicted relationships. Here, we used whole-genome sequence data from dove parasites (71 feather louse species of the genus Columbicola) and phylogenetic comparative methods to study the relationship between parasite effective population size and host population size and body size. We found that parasite effective population size is largely explained by host body size but not host population size. These results suggest the potential local population size (infrapopulation or deme size) is more predictive of the long-term effective population size of parasites than is the total number of potential parasite infrapopulations (i.e., host individuals).

Estimating parasite infrapopulation size given imperfect detection: Proof-of-concept with ectoparasitic fleas on prairie dogs.

Parasite infrapopulation size - the population of parasites affecting a single host - is a central metric in parasitology. However, parasites are small and elusive such that imperfect detection is expected. Repeated sampling of parasites during primary sampling occasions (e.g., each host capture) informs the detection process. Here, we estimate flea (Siphonaptera) infrapopulation size on black-tailed prairie dogs (Cynomys ludovicianus, BTPDs) as a proof-of-concept for estimating parasite infrapopulations given imperfect detection. From Jun-Aug 2011, we live-trapped 299 BTPDs for a total of 573 captures on 20 plots distributed among 13 colonies at the Vermejo Park Ranch, New Mexico, USA. During each capture, an anesthetized BTPD was combed 3 times consecutively, 15s each, to remove and count fleas. Each flea (n=4846) was linked to the BTPD from which it was collected and assigned an encounter history ('100', '010', '001'). We analyzed the encounter histories using Huggins closed captures models, setting recapture probabilities to 0, thereby accounting for flea removal from hosts. The probability of detecting an individual flea (p) increased with Julian date; field personnel may have become more efficient at combing fleas as the field season progressed. Combined p across 3 combings equaled 0.99. Estimates of flea infrapopulation size were reasonable and followed the negative binomial distribution. Our general approach may be broadly applicable to estimating infrapopulation sizes for parasites. The utility of this approach increases as p declines but, if p is very low, inference is likely limited.

High levels of inbreeding with spatial and host-associated structure in lice of an endangered freshwater seal.

Host‐specialist parasites of endangered large vertebrates are in many cases more endangered than their hosts. In particular, low host population densities and reduced among‐host transmission rates are expected to lead to inbreeding within parasite infrapopulations living on single host individuals. Furthermore, spatial population structures of directly‐transmitted parasites should be concordant with those of their hosts. Using population genomic approaches, we investigated inbreeding and population structure in a host‐specialist seal louse (Echinophthirius horridus) infesting the Saimaa ringed seal (Phoca hispida saimensis), which is endemic to Lake Saimaa in Finland, and is one of the most endangered pinnipeds in the world. We conducted genome resequencing of pairs of lice collected from 18 individual Saimaa ringed seals throughout the Lake Saimaa complex. Our analyses showed high genetic similarity and inbreeding between lice inhabiting the same individual seal host, indicating low among‐host transmission rates. Across the lake, genetic differentiation among individual lice was correlated with their geographic distance, and assignment analyses revealed a marked break in the genetic variation of the lice in the middle of the lake, indicating substantial population structure. These findings indicate that movements of Saimaa ringed seals across the main breeding areas of the fragmented Lake Saimaa complex may in fact be more restricted than suggested by previous population‐genetic analyses of the seals themselves.

Somatic Dimorphism in Cercariae of a Bird Schistosome.

Phenotypic polymorphism is a commonly observed phenomenon in nature, but extremely rare in free-living stages of parasites. We describe a unique case of somatic polymorphism in conspecific cercariae of the bird schistosome Trichobilharzia sp. “peregra”, in which two morphs, conspicuously different in their size, were released from a single Radix balthica snail. A detailed morphometric analysis that included multiple morphological parameters taken from 105 live and formalin-fixed cercariae isolated from several naturally infected snails provided reliable evidence for a division of all cercariae into two size groups that contained either large or small individuals. Large morph (total body length of 1368 and 1339 μm for live and formalin-fixed samples, respectively) differed significantly nearly in all morphological characteristics compared to small cercariae (total body length of 976 and 898 μm for live and formalin samples, respectively), regardless of the fixation method. Furthermore, we observed that small individuals represent the normal/commonly occurring phenotype in snail populations. The probable causes and consequences of generating an alternative, much larger phenotype in the parasite infrapopulation are discussed in the context of transmission ecology as possible benefits and disadvantages facilitating or preventing the successful completion of the life cycle.

Metazoan Parasite Infracommunities of the Tomtate Grunt (Haemulon aurolineatum) as Potential Bioindicators of Environmental Conditions in the Yucatan Continental Shelf, Mexico.

Metazoan parasites and their hosts experience the environmental influence of the seasonal presence of Western and Eastern subzones with different oceanographic conditions on the Yucatan continental shelf (YS). In addition, natural seeps and oil transport in the area lead to the presence of hydrocarbons. We hypothesized that the parasite infrapopulations and infracommunities of Haemulon aurolineatum will respond to environmental variability related with ongoing oceanographic subzone conditions of the YS. Spatial and multivariate statistical analyses were used to test this hypothesis. For 17 sampling sites along the YS, 55 parasite morphospecies were recovered from 146 fishes. There were significant differences in the number of parasite species between Western and Eastern subzones, but not in the number of parasite individuals. Spatial autocorrelation on environmental variables as a consequence of the Yucatan current was found. Overall, the parasite metrics suggested a region with a good ecosystem health condition naturally influenced by oceanographic processes.

Patterns of Microbiome Variation Among Infrapopulations of Permanent Bloodsucking Parasites.

While interspecific variation in microbiome composition can often be readily explained by factors such as host species identity, there is still limited knowledge of how microbiomes vary at scales lower than the species level (e.g., between individuals or populations). Here, we evaluated variation in microbiome composition of individual parasites among infrapopulations (i.e., populations of parasites of the same species living on a single host individual). To address this question, we used genome-resolved and shotgun metagenomic data of 17 infrapopulations (balanced design) of the permanent, bloodsucking seal louse Echinophthirius horridus sampled from individual Saimaa ringed seals Pusa hispida saimensis. Both genome-resolved and read-based metagenomic classification approaches consistently show that parasite infrapopulation identity is a significant factor that explains both qualitative and quantitative patterns of microbiome variation at the intraspecific level. This study contributes to the general understanding of the factors driving patterns of intraspecific variation in microbiome composition, especially of bloodsucking parasites, and has implications for understanding how well-known processes occurring at higher taxonomic levels, such as phylosymbiosis, might arise in these systems.

Genetic evidence for the role of non-human primates as reservoir hosts for human schistosomiasis.

BackgroundSchistosomiasis is a chronic parasitic disease, that affects over 207 million people and causes over 200,000 deaths annually, mainly in sub-Saharan Africa. Although many health measures have been carried out to limit parasite transmission, significant numbers of non-human primates such as Chlorocebus aethiops (Ch. aethiops) (vervet) and Papio anubis (baboon) are infected with S. mansoni, notably in Ethiopia, where they are expected to have potentially significant implications for transmission and control efforts.ObjectiveThe objective of this study was to assess and compare the genetic diversity and population structure of S. mansoni isolates from human and non-human primates free-ranging in close proximity to villages in selected endemic areas of Ethiopia.MethodsA cross-sectional study was conducted in three transmission sites: Bochesa, Kime and Fincha. A total of 2,356 S. mansoni miracidia were directly isolated from fecal specimens of 104 hosts (i.e. 60 human hosts and 44 non-human primates). We performed DNA extraction and PCR amplification using fourteen microsatellite loci.ResultsAt population scale we showed strong genetic structure between the three sample sites. At the definitive host scale, we observed that host factors can shape the genetic composition of parasite infra-populations. First, in male patients, we observed a positive link between parasite genetic diversity and the age of the patients. Second, we observed a difference in genetic diversity which was high in human males, medium in human females and low in non-human primates (NHPs). Finally, whatever the transmission site no genetic structure was observed between human and non-human primates, however, there appears to be little barriers, if any, host specificity of the S. mansoni populations with cross-host infections.ConclusionOccurrence of infection of a single host with multiple S. mansoni strains and inter- and intra-host genetic variations was observed. Substantial genetic diversity and gene flow across the S. mansoni population occurred at each site and non-human primates likely play a role in local transmission and maintenance of infection. Therefore, public health and wildlife professionals should work together to improve disease control and elimination strategies.

Habitat selection and populations of Corynosoma (Acanthocephala) in the intestines of sea otters (Enhydra lutris) and seals.

Acanthocephalans are common intestinal parasites of marine mammals, the most widespread of which is the genus Corynosoma. In this study, parasite infrapopulations of two closely related species of Corynosoma were examined: Corynosoma enhydri from sea otters (Enhydra lutris) in Alaska (n = 12) and California (n = 19), and Corynosoma strumosum from seals in Germany (n = 22). Prevalence of C. enhydri was 100% in Californian otters, with a mean abundance of 30, and 83% in Alaskan otters, with a mean abundance of 232. In seals, C. strumosum had a prevalence of 65%, with a mean abundance of 33. Female C. enhydri dominated both Californian (82%) and Alaskan (79%) infections, while, in seals, female C. strumosum made up 68% of the parasite population. Reproduction rates for C. enhydri, with 16% (California) and 18% (Alaska) of females mated, were low compared to C. strumosum in seals, of which 40% of females were mated. Habitat selection also differed significantly between the two species. Corynosoma enhydri was found most frequently in the second and third fifths of the small intestine, while C. strumosum was found most frequently in the fourth. The differences in habitat selection and prevalence analysed in this study may be related to a trade-off between growth and reproduction between the two species.

Microhabitat Use of Allocreadium lobatum (Trematoda) in Creek Chub (Semotilus atromaculatus) in Southeastern Nebraska

Microhabitat use of Allocreadium lobatum (Trematoda) in the intestine of creek chub (Semotilus atromaculatus) was studied using natural infections. Seventy-six creek chub harboring only A. lobatum in their intestine provided data on the position of 212 worms along the length of the intestine. Positions of worms within the intestine co-varied strongly with host size and therefore intestinal length; worm positions standardized by intestinal length eliminated this relationship. Analyses of standardized worm positions (worm location) revealed that microhabitat use did not vary with worm size, maturity, or number of eggs in the uterus. Individuals of A. lobatum occurred throughout the intestine, but were concentrated in the posterior 30% of the intestine, regardless of demographic properties of the worms themselves. Standardized niche breadths of parasite infrapopulations suggested strict microhabitat specificity, but nearest-neighbor analyses revealed very little aggregation of worms in infrapopulations beyond what is expected by chance. Overall, trematodes of this species appear to exhibit some microhabitat specificity when examined with snapshot, field-collected data, but there is no clear indication of what mechanism might produce the observed pattern. Use of the intestine by individuals of A. lobatum might be more dynamic than can be revealed by the present investigation or studies similar to it.

Host B chromosomes as potential sex ratio distorters of intestinal nematode infrapopulations in the yellow-necked mouse (Apodemus flavicollis).

The yellow-necked mouse, Apodemus flavicollis, can be considered as a model for genetic polymorphism produced by the frequent presence of supernumerary or B chromosomes (Bs). Host genetic background is rarely taken into account in studies of parasite sex ratio. The main aim of this study was to investigate the range of infrapopulation sex ratios for nematode parasites of the yellow-necked mouse and to determine which factors most influence variation in parasite sex ratios. Six nematode species found in the collected yellow-necked mice were analysed. We confirmed the predominant pattern of female-biased sex ratios in vertebrate parasite infrapopulations. The presence of B chromosomes in host genomes played an important role in infrapopulations of Heligmosomoides polygyrus, Syphacia stroma and Trichuris muris, as hosts with B chromosomes carried a higher proportion of males. The relative increase of males in infrapopulations could result from a shift in parasite life history strategy, induced by adaptation to the specific host genotypes (Bs present). In a meta-analysis with previously published data, the sex determination system was demonstrated to play a significant role in nematode sex ratio variation, as well as specific life history patterns, such as the place of egg hatching.

Positive density-dependent growth supports costs sharing hypothesis and population density sensing in a manipulative parasite.

Parasites manipulate their hosts' phenotype to increase their own fitness. Like any evolutionary adaptation, parasitic manipulations should be costly. Though it is difficult to measure costs of the manipulation directly, they can be evaluated using an indirect approach. For instance, theory suggests that as the parasite infrapopulation grows, the investment of individual parasites in host manipulation decreases, because of cost sharing. Another assumption is that in environments where manipulation does not pay off for the parasite, it can decrease its investment in the manipulation to save resources. We experimentally infected rainbow trout Oncorhynchus mykiss with the immature larvae of the trematode Diplostomum pseudospathaceum, to test these assumptions. Immature D. pseudospathaceum metacercariae are known for their ability to manipulate the behaviour of their host enhancing its anti-predator defenses to avoid concomitant predation. We found that the growth rate of individual parasites in rainbow trout increased with the infrapopulation size (positive density-dependence) suggesting cost sharing. Moreover, parasites adjusted their growth to the intensity of infection within the eye lens where they were localized suggesting population density sensing. Results of this study support the hypothesis that macroparasites can adjust their growth rate and manipulation investment according to cost sharing level and infrapopulation size.

A stochastic model to investigate the effects of control strategies on calves exposed to Ostertagia ostertagi.

Predicting the effectiveness of parasite control strategies requires accounting for the responses of individual hosts and the epidemiology of parasite supra- and infra-populations. The first objective was to develop a stochastic model that predicted the parasitological interactions within a group of first season grazing calves challenged by Ostertagia ostertagi, by considering phenotypic variation amongst the calves and variation in parasite infra-population. Model behaviour was assessed using variations in parasite supra-population and calf stocking rate. The model showed the initial pasture infection level to have little impact on parasitological output traits, such as worm burdens and FEC, or overall performance of calves, whereas increasing stocking rate had a disproportionately large effect on both parasitological and performance traits. Model predictions were compared with published data taken from experiments on common control strategies, such as reducing stocking rates, the 'dose and move' strategy and strategic treatment with anthelmintic at specific times. Model predictions showed in most cases reasonable agreement with observations, supporting model robustness. The stochastic model developed is flexible, with the potential to predict the consequences of other nematode control strategies, such as targeted selective treatments on groups of grazing calves.

Seasonal profile of metal accumulation in the acanthocephalan Pomphorhynchus laevis: a valuable tool to study infection dynamics and implications for metal monitoring

BackgroundA large number of studies demonstrated that acanthocephalans exhibit a high metal accumulation potential and thus can be used as sensitive accumulation indicators. However, similar to free-living bioindicators, a seasonal variation in metal concentrations in parasites might occur. Accordingly, the influence of seasonality has to be elucidated if parasites should be applied as sentinels.MethodsIn order to assess a possible seasonal profile of element concentrations, the concentrations of As, Cd, Co, Cu, Fe, Mn, Mo, Ni, Pb, V and Zn in the acanthocephalan Pomphorhynchus laevis and in its host barbel (Barbus barbus) were analysed in a seasonal manner (spring, summer and autumn) using inductively coupled plasma mass spectrometry (ICP-MS).ResultsFive elements (As, Cd, Cu, Pb and Zn) were detected in significantly higher concentrations in the parasites compared to host muscle, intestine and liver. Their levels in P. laevis showed a clear seasonal pattern, while the concentrations in the fish tissues remained similar during the year. The highest concentrations in the parasites were found in autumn, followed by spring and summer. Evidence from the literature suggests that this profile coincides with the seasonality of acanthocephalan transmission, as their annual concentration profile reflected the mean individual weight pattern during the year. Parasite infrapopulations in autumn consisted mainly of young worms which are characterised by an accelerated metabolism and a higher surface to volume ratio resulting in higher element concentrations when compared to older worms which are assumed to slow down their metabolism and additionally excrete metals with their eggs.ConclusionsBased on the available data from the present study and literature, a model is suggested, which visualises the accumulation kinetic of several elements under natural conditions. According to the element accumulation data the lifespan of P. laevis in barbel was roughly estimated to range between six and eight months.

Sexual segregation of Echinorhynchus borealis von Linstow, 1901 (Acanthocephala) in the gut of burbot (Lota lota Linnaeus).

Helminths often occupy defined niches in the gut of their definitive hosts. In the dioecious acanthocephalans, adult males and females usually have similar gut distributions, but sexual site segregation has been reported in at least some species. We studied the intestinal distribution of the acanthocephalan Echinorhynchus borealis von Linstow, 1901 (syn. of E. cinctulus Porta, 1905) in its definitive host, burbot (Lota lota Linnaeus). Over 80% of female worms were found in the pyloric caeca, whereas the majority of males were in the anterior two-thirds of the intestine. This difference was relatively consistent between individual fish hosts. Worms from different parts of the gut did not differ in length, so site segregation was not obviously related to worm growth or age. We found proportionally more males in the caeca when a larger fraction of the females were found there, suggesting mating opportunities influence gut distribution. However, this result relied on a single parasite infrapopulation and is thus tentative. We discuss how mating strategies and/or sexual differences in life history might explain why males and females occupy different parts of the burbot gut.

Lineage sorting in multihost parasites: Eidmanniella albescens and Fregatiella aurifasciata on seabirds from the Galapagos Islands.

Parasites comprise a significant percentage of the biodiversity of the planet and are useful systems to test evolutionary and ecological hypotheses. In this study, we analyze the effect of host species identity and the immediate local species assemblage within mixed species colonies of nesting seabirds on patterns of genetic clustering within two species of multihost ectoparasitic lice. We use three genetic markers (one mitochondrial, COI, and two nuclear, EF1-α and wingless) and maximum likelihood phylogenetic trees to test whether (1) parasites show lineage sorting based on their host species; and (2) switching of lineages to the alternate host species depends on the immediate local species assemblage of individual hosts within a colony. Specifically, we examine the genetic structure of two louse species: Eidmanniella albescens, infecting both Nazca (Sula granti) and blue-footed boobies (Sula nebouxii), and Fregatiella aurifasciata, infecting both great (Fregata minor) and magnificent frigatebirds (Fregata magnificens). We found that host species identity was the only factor explaining the patterns of genetic structure in both parasites. In both cases, there is evident genetic differentiation depending on the host species. Thus, a revision of the taxonomy of these louse species is needed. One possible explanation of this pattern is extremely low louse migration rates between host species, perhaps influenced by fine-scale spatial separation of host species within mixed colonies, and low parasite infrapopulation numbers.

Drivers of aggregation in a novel arboreal parasite: the influence of host size and infra-populations

As a novel arboreal parasite, New Zealand’s largest endemic moth, Aenetus virescens, is a biological oddity. With arguably the most unusual lepidopteran life history on earth, larvae grow to 100mm, spending ∼6years as wood-boring parasites feeding on host tree phloem. Parasite fitness is a product of host suitability. Parasite discrimination between heterogeneous hosts in fragmented populations shapes parasite aggregation. We investigated whether A. virescens aggregation among hosts occurs randomly (target area effect), or if larvae select hosts based on host quality (ideal free distribution). Using long-term larval growth as an indicator of energy intake, we examined A. virescens aggregation in relation to host size and infra-population. Using a generalised linear model, the relationship between parasite intensity and host tree size was analysed. Reduced major axis regression was used to evaluate A. virescens growth after 1year. Linear mixed-effects models inferred the influence of parasite infra-population on parasite growth, with host tree as a random factor. Results indicate parasite intensity scaled positively with host size. Furthermore, parasite growth remained consistent throughout ontogeny regardless of host size or parasite infra-population. Aenetus virescens aggregation among hosts violates the ideal free distribution hypothesis, occurring instead as a result of host size, supporting the target area effect.

Parasite community of Pagrus pagrus (Sparidae) from Rio de Janeiro, Brazil: evidence of temporal stability.

One hundred specimens of Pagrus pagrus were necropsied for parasitological study between January and February 2012. These data were compared with data from 90 specimens that had been examined in the year 2000. The fish were collected from Cabo Frio (21-23 ° S, 41-45 ° W), in the state of Rio de Janeiro, Brazil. In the current study a total of 19 parasite species were collected. Sixteen of these were collected in the samples of both 2012 and 2000, thus totaling 25 parasite species in the two studies. All the results were similar to those of the previous study with regard to: dominance of anisakid nematode larvae; correlation between abundance of parasites and host body size; absence of correlations between the size of parasite infrapopulations and host sex; and scarcity of interspecific associations. The only difference between the studies was in the abundance values for Anisakidae, which could be correlated with the seasonal factor of the collection period, which was concentrated in the summer. This is also the period of the South Atlantic Central Water (SACW) phenomenon, during which there is greater host reproductive activity. Therefore, significant stability of the metazoan community structure of P. pagrus was shown.

In vivo albendazole treatment of Taenia crassiceps cysticerci strain WFU: proliferation, damage, and recovery

Taenia crassiceps has been widely experimented as a model for in vitro and in vivo studies on drug responses. The purpose of this study was to treat BALB/c mice infected with T. crassiceps strain WFU with commercially available albendazole and to analyze the reduction in parasite infrapopulations. Here, we describe the reduction and apparent damage of T. crassicceps WFU cysticerci in infected mice after antihelminthic drug treatment and subsequent inoculation of those treated parasites into a naïve host. We were able to reduce significantly the parasite counts to 33 and 48% after albendazole treatment for 20 or 25 days and compared with the untreated mice. We also observed morphological damage such as the partial blebbing in the tegument and parenchyma of treated parasites, as well as disorganized musculature and the loss of cell membranes in subtegumental tissue section. However, larvae from albendazole-treated mice inoculated into the next host were able to become re-established in the next murine host due, probably, to the survival of proliferative parasite cells.