Asian Parasite Research Articles

The potential effects of climate change on the climatic suitability patterns of the Western Asian vectors and parasites of cutaneous leishmaniasis in the mid- and late twenty-first century

Climate change alters the global distribution of leishmaniasis vector sandfly species. However, the possible effect of altering environmental factors on sandfly species varies by species and area. The aim of this study was to project the future potential effect of climate change on the Western Asian occurrence of the sandfly vector species Phlebotomus papatasi and Phlebotomus sergenti and the parasites Leishmania major and Leishmania tropica for 2041–2060 and 2081–2100 compared to their modelled climatic suitability patterns in the reference period 1970–2000. The model results suggest that by 2041–2060 and 2081–2100, in the coastal areas of the Levantine countries and the mid-elevation regions of Western Iran and Southeast Turkey, the climatic suitability of all the studied species is predicted to increase. In contrast, the model results suggest the decline of the populations of the studied vectors as well as the disappearance of the parasites in Iraq, North Saudi Arabia, Kuwait, and the inland regions of Western Asian countries. Considering the present-day population density patterns, it can be said that although in large regions of the region, the climatic suitability values are predicted to decrease, the populated Levantine regions seem to be at risk of climate change-facilitated increase in cutaneous leishmaniasis in the second half of the twenty-first century. The model results suggest that climate change will especially increase the climatic suitability of Leishmania tropica in the Levantine region in the late twenty-first century.

A Malaria Parasite Cross Reveals Genetic Determinants of Plasmodium falciparum Growth in Different Culture Media.

What genes determine in vitro growth and nutrient utilization in asexual blood-stage malaria parasites? Competition experiments between NF54, clone 3D7, a lab-adapted African parasite, and a recently isolated Asian parasite (NHP4026) reveal contrasting outcomes in different media: 3D7 outcompetes NHP4026 in media containing human serum, while NHP4026 outcompetes 3D7 in media containing AlbuMAX, a commercial lipid-rich bovine serum formulation. To determine the basis for this polymorphism, we conducted parasite genetic crosses using humanized mice and compared genome-wide allele frequency changes in three independent progeny populations cultured in media containing human serum or AlbuMAX. This bulk segregant analysis detected three quantitative trait loci (QTL) regions [on chromosome (chr) 2 containing aspartate transaminase AST; chr 13 containing EBA-140; and chr 14 containing cysteine protease ATG4] linked with differential growth in serum or AlbuMAX in each of the three independent progeny pools. Selection driving differential growth was strong (s = 0.10 – 0.23 per 48-hour lifecycle). We conducted validation experiments for the strongest QTL on chr 13: competition experiments between ΔEBA-140 and 3D7 wildtype parasites showed fitness reversals in the two medium types as seen in the parental parasites, validating this locus as the causative gene. These results (i) demonstrate the effectiveness of bulk segregant analysis for dissecting fitness traits in P. falciparum genetic crosses, and (ii) reveal intimate links between red blood cell invasion and nutrient composition of growth media. Use of parasite crosses combined with bulk segregant analysis will allow systematic dissection of key nutrient acquisition/metabolism and red blood cell invasion pathways in P. falciparum.

Plasmodium falciparum K13 mutations in Africa and Asia impact artemisinin resistance and parasite fitness.

The emergence of mutant K13-mediated artemisinin (ART) resistance in Plasmodium falciparum malaria parasites has led to widespread treatment failures across Southeast Asia. In Africa, K13-propeller genotyping confirms the emergence of the R561H mutation in Rwanda and highlights the continuing dominance of wild-type K13 elsewhere. Using gene editing, we show that R561H, along with C580Y and M579I, confer elevated in vitro ART resistance in some African strains, contrasting with minimal changes in ART susceptibility in others. C580Y and M579I cause substantial fitness costs, which may slow their dissemination in high-transmission settings, in contrast with R561H that in African 3D7 parasites is fitness neutral. In Cambodia, K13 genotyping highlights the increasing spatio-temporal dominance of C580Y. Editing multiple K13 mutations into a panel of Southeast Asian strains reveals that only the R561H variant yields ART resistance comparable to C580Y. In Asian Dd2 parasites C580Y shows no fitness cost, in contrast with most other K13 mutations tested, including R561H. Editing of point mutations in ferredoxin or mdr2, earlier associated with resistance, has no impact on ART susceptibility or parasite fitness. These data underline the complex interplay between K13 mutations, parasite survival, growth and genetic background in contributing to the spread of ART resistance.

The power and promise of genetic mapping from Plasmodium falciparum crosses utilizing human liver-chimeric mice

Genetic crosses are most powerful for linkage analysis when progeny numbers are high, parental alleles segregate evenly and numbers of inbred progeny are minimized. We previously developed a novel genetic crossing platform for the human malaria parasite Plasmodium falciparum, an obligately sexual, hermaphroditic protozoan, using mice carrying human hepatocytes (the human liver-chimeric FRG NOD huHep mouse) as the vertebrate host. We report on two genetic crosses—(1) an allopatric cross between a laboratory-adapted parasite (NF54) of African origin and a recently patient-derived Asian parasite, and (2) a sympatric cross between two recently patient-derived Asian parasites. We generated 144 unique recombinant clones from the two crosses, doubling the number of unique recombinant progeny generated in the previous 30 years. The allopatric African/Asian cross has minimal levels of inbreeding and extreme segregation distortion, while in the sympatric Asian cross, inbred progeny predominate and parental alleles segregate evenly. Using simulations, we demonstrate that these progeny provide the power to map small-effect mutations and epistatic interactions. The segregation distortion in the allopatric cross slightly erodes power to detect linkage in several genome regions. We greatly increase the power and the precision to map biomedically important traits with these new large progeny panels.

Distinctive genetic structure and selection patterns in Plasmodium vivax from South Asia and East Africa

Despite the high burden of Plasmodium vivax malaria in South Asian countries, the genetic diversity of circulating parasite populations is not well described. Determinants of antimalarial drug susceptibility for P. vivax in the region have not been characterised. Our genomic analysis of global P. vivax (n = 558) establishes South Asian isolates (n = 92) as a distinct subpopulation, which shares ancestry with some East African and South East Asian parasites. Signals of positive selection are linked to drug resistance-associated loci including pvkelch10, pvmrp1, pvdhfr and pvdhps, and two loci linked to P. vivax invasion of reticulocytes, pvrbp1a and pvrbp1b. Significant identity-by-descent was found in extended chromosome regions common to P. vivax from India and Ethiopia, including the pvdbp gene associated with Duffy blood group binding. Our investigation provides new understanding of global P. vivax population structure and genomic diversity, and genetic evidence of recent directional selection in this important human pathogen.

Khawia japonensis (Cestoda), the Asian parasite of common carp, continues to spread in Central European countries: distribution, infection indices and histopathology

Khawia japonensis (Cestoda), the Asian parasite of common carp, continues to spread in Central European countries: distribution, infection indices and histopathology

Ookinete-Specific Genes and 18S SSU rRNA Evidenced in Plasmodium vivax Selection and Adaptation by Sympatric Vectors.

In the southern Pacific coast of Chiapas, Mexico (SM), the two most abundant vector species, Nyssorhynchus albimanus and Anopheles pseudopunctipennis, were susceptible to different Plasmodium vivax Pvs25/28 haplotypes. To broaden our understanding of the existing P. vivax in the area, genes encoding proteins relevant for ookinete development and the 18S rRNA were studied. P. vivax infectivity (percentage of infected mosquitoes and oocyst numbers) was evaluated by simultaneously feeding infected blood samples from patients to Ny. albimanus and An. pseudopunctipennis female mosquitoes. Three infectivity patterns were identified: one group of parasites were more infective to An. pseudopunctipennis than to Ny. albimanus, another group was more infective to Ny. albimanus, while a third group infected both vectors similarly. In 29 parasite isolates, the molecular variations of ookinete-specific genes and the 18S rRNA-type S were analyzed. Using concatenated sequences, phylogenetic trees, and Structure analysis, parasite clustering within SM isolates and between these and those from other geographical origins were investigated. A ML phylogenetic tree resolved two parasite lineages: PvSM-A and PvSM-B. They were associated to a different 18S rRNA variant. PvSM-A parasites had 18S rRNA variant rV2 and correspond to parasites causing high oocyst infection in Ny. albimanus. A new ML tree and Structure analysis, both comprising global sequences, showed PvSM-A clustered with Latin American parasites. Meanwhile, all isolates of PvSM-B had 18S rRNA variant rV1 and remained as unique genetic cluster comprising two subgroups: PvSM-Ba, producing high infection in An. pseudopunctipennis, and PvSM-Bb, causing similar oocyst infection in both vector species. PvSM-A parasites were genetically similar to parasites from South America. Meanwhile, PvSM-B were exclusive to southern Mexico and share ancestry with Asian parasites. The results suggest that these lineages evolved separately, likely by geographic and vector restriction.

Analysis of Plasmodium falciparum Rh2b deletion polymorphism across different transmission areas

Despite significant progress in controlling malaria, the disease remains a global health burden. The intricate interactions the parasite Plasmodium falciparum has with its host allows it to grow and multiply in human erythrocytes. The mechanism by which P. falciparum merozoites invade human erythrocytes is complex, involving merozoite proteins as well as erythrocyte surface proteins. Members of the P. falciparum reticulocyte binding-like protein homolog (PfRh) family of proteins play a pivotal role in merozoite invasion and hence are important targets of immune responses. Domains within the PfRh2b protein have been implicated in its ability to stimulate natural protective antibodies in patients. More specifically, a 0.58 kbp deletion, at the C-terminus has been reported in high frequencies in Senegalese and Southeast Asian parasite populations, suggesting a possible role in immune evasion. We analysed 1218 P. falciparum clinical isolates, and the results show that this deletion is present in Ghanaian parasite populations (48.5% of all isolates), with Kintampo (hyper-endemic, 53.2%), followed by Accra (Hypo-endemic, 50.3%), Cape Coast (meso-endemic, 47.9%) and Sogakope (meso-endemic, 43.15%). Further analysis of parasite genomes stored in the MalariaGEN database revealed that the deletion variant was common across transmission areas globally, with an overall frequency of about 27.1%. Interestingly, some parasite isolates possessed mixed PfRh2b deletion and full-length alleles. We further showed that levels of antibodies to the domain of PfRh2 protein were similar to antibody levels of PfRh5, indicating it is less recognized by the immune system.

Apicoplast phylogeny reveals the position of Plasmodium vivax basal to the Asian primate malaria parasite clade

The malaria parasite species, Plasmodium vivax infects not only humans, but also African apes. Human specific P. vivax has evolved from a single ancestor that originated from a parasite of African apes. Although previous studies have proposed phylogenetic trees positioning P. vivax (the common ancestor of human and African ape P. vivax) within the assemblages of Asian primate parasites, its position has not yet been robustly confirmed. We determined nearly complete apicoplast genome sequences from seven Asian primate parasites, Plasmodium cynomolgi (strains Ceylonensis and Berok), P. knowlesi P. fragile, P. fieldi, P. simiovale, P. hylobati, P. inui, and an African primate parasite, P. gonderi, that infects African guenon. Phylogenetic relationships of the Plasmodium species were analyzed using newly and previously determined apicoplast genome sequences. Multigene maximum likelihood analysis of 30 protein coding genes did not position P. vivax within the Asian primate parasite clade but positioned it basal to the clade, after the branching of an African guenon parasite, P. gonderi. The result does not contradict with the emerging notion that P. vivax phylogenetically originated from Africa. The result is also supported by phylogenetic analyses performed using massive nuclear genome data of seven primate Plasmodium species.

A new pathogen spillover from domestic to wild animals: Plasmodium juxtanucleare infects free-living passerines in Brazil.

Habitat modification may facilitate the emergence of novel pathogens, and the expansion of agricultural frontiers make domestic animals important sources of pathogen spillover to wild animals. We demonstrate for the first time that Plasmodium juxtanucleare, a widespread parasite from domestic chickens, naturally infects free-living passerines. We sampled 68 wild birds within and at the border of conservation units in central Brazil composed by Cerrado, a highly threatened biome. Seven out of 10 passerines captured in the limits of a protected area with a small farm were infected by P. juxtanucleare as was confirmed by sequencing a fragment of the parasite's cytochrome b. Blood smears from these positive passerines presented trophozoites, meronts and gametocytes compatible with P. juxtanucleare, meaning these birds are competent hosts for this parasite. After these intriguing results, we sampled 30 backyard chickens managed at the area where P. juxtanucleare-infected passerines were captured, revealing one chicken infected by the same parasite lineage. We sequenced the almost complete mitochondrial genome from all positive passerines, revealing that Brazilian and Asian parasites are closely related. P. juxtanucleare can be lethal to non-domestic hosts under captive and rehabilitation conditions, suggesting that this novel spillover may pose a real threat to wild birds.

Updating Taenia asiatica in humans and pigs.

An epidemiological study on taeniasis and cysticercosis in northern India has recently updated the epidemiology of Taenia asiatica. Practically, all the detected cases of taeniasis were caused by T. asiatica, cited for the first time in humans in that country. The finding widens the geographical distribution of T. asiatica, a species wrongly considered an exclusive South-Eastern Asian parasite. Due to the introduction of molecular techniques in Taenia diagnosis, the species is slowly showing its true distribution. A human Taenia species with cosmopolitan hosts (the same as the other two Taenia species) but limited to a specific geographical area and not affected by globalisation would certainly be hard to believe. Regarding cysticercosis, there is a remarkable finding concerning T. asiatica pig cysticercosis, specifically the presence of the cysticercus of T. asiatica not only in the liver (its preferential infection site) but also in muscle. This is the first time that the cysticercus of T. asiatica has been found in muscle in a naturally infected pig. This fact is actually relevant since people are at a greater risk of becoming infected by T. asiatica than previously expected since the liver is no longer the only site of pig infection. The Taenia species causing Taenia saginata-like taeniasis around the world, as well as pig and human cysticercosis, should always be molecularly confirmed since T. asiatica could be involved.

Evidence of a Mild Mutator Phenotype in Cambodian Plasmodium falciparum Malaria Parasites.

Malaria control efforts have been continuously stymied by drug-resistant strains of Plasmodium falciparum, which typically originate in Southeast Asia prior to spreading into high-transmission settings in Africa. One earlier proposed explanation for Southeast Asia being a hotbed of resistance has been the hypermutability or “Accelerated Resistance to Multiple Drugs” (ARMD) phenotype, whereby multidrug-resistant Southeast Asian parasites were reported to exhibit 1,000-fold higher rates of resistance to unrelated antimalarial agents when compared to drug-sensitive parasites. However, three recent studies do not recapitulate this hypermutability phenotype. Intriguingly, genome sequencing of recently derived multidrug-resistant Cambodian isolates has identified a high proportion of DNA repair gene mutations in multidrug-resistant parasites, suggesting their potential role in shaping local parasite evolution. By adapting fluctuation assays for use in P. falciparum, we have examined the in vitro mutation rates of five recent Cambodian isolates and three reference laboratory strains. For these studies we also generated a knockout parasite line lacking the DNA repair factor Exonuclease I. In these assays, parasites were typed for their ability to acquire resistance to KAE609, currently in advanced clinical trials, yielding 13 novel mutations in the Na+/H+-ATPase PfATP4, the primary resistance determinant. We observed no evidence of hypermutability. Instead, we found evidence of a mild mutator (up to a 3.4-fold increase in mutation rate) phenotype in two artemisinin-resistant Cambodian isolates, which carry DNA repair gene mutations. We observed that one such mutation in the Mismatch Repair protein Mlh1 contributes to the mild mutator phenotype when modeled in yeast (scmlh1-P157S). Compared to basal rates of mutation, a mild mutator phenotype may provide a greater overall benefit for parasites in Southeast Asia in terms of generating drug resistance without incurring detrimental fitness costs.

Comment on "Epidemiological Survey on Porcine Cysticercosis in Nay Pyi Taw Area, Myanmar".

We have read with interest the paper by Khaing et al. [1], in which first data on Taenia solium cysticercosis in pigs from Myanmar are published. The authors found a porcine cysticercosis prevalence of 23.67% in slaughtered pigs, which, as they mention, indicates the presence of human taeniasis and also the risk of acquiring human cysticercosis and, therefore, neurocysticercosis. The high porcine cysticercosis prevalence detected by the authors means, obviously, that there has to be a high prevalence of human T. solium taeniasis among the inhabitants of Myanmar leading to a high presence of infective eggs in the environment. However, as far as we know, the presence of T. solium taeniasis, as well as neurocysticercosis, has only been diagnosed in refugees or immigrants from that country but not in people living in Myanmar [2–6]. There is no doubt that T. solium is present in Myanmar; however, our query is whether other human Taenia species might also be present in that country and, in particular, whether T. asiatica could be present in Myanmar as suggested more than 20 years ago by Fan et al. [7] and recently by Anantaphruti [2]. T. asiatica was described as a new species in 1993 [8]. This third human Taenia species has practically the same gravid proglottid morphology as Taenia saginata, but its life cycle is just the same as that of T. solium; that is, its intermediate hosts are pigs instead of cattle, the intermediate hosts for T. saginata. The clear liver tropism the cysticerci of T. asiatica presents in pigs is worth mentioning. Regarding its geographical distribution, the species has been found in Taiwan, South Korea, Philippines, Indonesia, Thailand, China, Vietnam, Japan, and Nepal [9]. Although T. asiatica was initially considered as an exclusively Southeastern Asian parasite, its finding in Nepal (far from Southeast Asia) and the phenomenon of globalization (it is a parasite with cosmopolitan hosts) have recently led to the notion that the species probably has a wider distribution [10, 11]. The presence of T. asiatica in three more Asian countries, Myanmar, Laos, and Malaysia, is highly suspected [9]. Concerning Myanmar, no cases have been confirmed so far, but a case identified as T. asiatica was detected in a Karen immigrant who moved from Myanmar to Kanchanaburi province, Thailand, in 2005. However, she had already started to expel proglottids in faeces in 1997, when she was still living in Myanmar [2]. Therefore, the presence of T. asiatica in Myanmar is very likely. T. asiatica may be found in the definitive (humans) as well as in the intermediatehost (pigs). The detection of T. asiatica taeniasis is by no means an easy task. Taking into account that neither the characteristics of the eggs nor the morphology of the gravid proglottids are specific enough to distinguish T. asiatica from T. saginata, these two species can only be differentiated by means of molecular techniques (e.g., multiplex PCR (polymerase chain reaction)) [12]. Unfortunately, these expensive molecular methods are not normally employed in routine diagnosis. A specific meat inspection should be carried out to detect T. asiatica cysticerci in pigs. Routine meat inspections conducted in pigs in Myanmar include the study of T. solium predilection sites such as the tongue, masseter, brain, shoulder, diaphragm, heart, and skeletal, fore limb as well as hind limb muscles [1]. Considering that the small cysticerci of T. asiatica are almost exclusively located in the liver, such local inspections would not be sensitive enough to detect infected livers [13]. In addition, T. asiatica cross-reacts even in the most specific immunological method to detect T. solium cysticercosis, the enzyme-linked immunoelectrotransfer blot (EITB) [14]. Therefore, serological tests would currently not be specific enough to detect T. asiatica cysticercosis. For the aforementioned reasons, we urge Khaing and colleagues to take advantage of their research in pigs in Myanmar and carry out a detailed study of the surface as well as the parenchyma of pig livers. T. asiatica cysticerci can easily be differentiated from those of T. solium due to their smaller size as well as the lack of hooklets or the presence of vestigial ones [8]. Molecular techniques, however, would be the key to ascertain whether T. asiatica is definitely present in Myanmar or not.

Susanlimae ianwhittingtoni gen. nov., sp. nov. (Monogenoidea: Dactylogyridae), a dweller of the gill rakers of Pseudeutropius moolenburghae (Siluriformes: Schilbeidae) from Sumatra

A new genus and species of Monogenoidea, Susanlimae ianwhittingtoni gen. nov., sp. nov., are proposed for dactylogyrids collected from the Nuayang tipis, Pseudeutropius moolenburghae Weber & de Beaufort, 1913, which inhabits freshwater in Sumatra. While clearly a member of a putative clade that includes Asian and African catfish parasites, S. ianwhittingtoni sp. nov. differs from most members of this clade by having a bifurcated haptor that embraces the gill rakers of its host. This haptoral morphology and mode of attachment also occur in species of Bifurcohaptor Jain, 1958. However, species of Susanlimae gen. nov. are easily distinguished by the comparative morphology of their haptoral armature. In Susanlimae gen. nov., the ventral and dorsal bars are single, elongated, and inverted u-shaped (ventral bar short and two short dorsal bars in Bifurcohaptor spp.); the dorsal anchor is robust with well-defined roots (reduced roots and elongate shaft in Bifurcohaptor spp.); and the ventral anchor has an elongated, deep root (inconspicuous in Bifurcohaptor spp.).

Pooled sequencing and rare variant association tests for identifying the determinants of emerging drug resistance in malaria parasites.

We explored the potential of pooled sequencing to swiftly and economically identify selective sweeps due to emerging artemisinin (ART) resistance in a South-East Asian malaria parasite population. ART resistance is defined by slow parasite clearance from the blood of ART-treated patients and mutations in the kelch gene (chr. 13) have been strongly implicated to play a role. We constructed triplicate pools of 70 slow-clearing (resistant) and 70 fast-clearing (sensitive) infections collected from the Thai–Myanmar border and sequenced these to high (∼150-fold) read depth. Allele frequency estimates from pools showed almost perfect correlation (Lin’s concordance = 0.98) with allele frequencies at 93 single nucleotide polymorphisms measured directly from individual infections, giving us confidence in the accuracy of this approach. By mapping genome-wide divergence (FST) between pools of drug-resistant and drug-sensitive parasites, we identified two large (>150 kb) regions (on chrs. 13 and 14) and 17 smaller candidate genome regions. To identify individual genes within these genome regions, we resequenced an additional 38 parasite genomes (16 slow and 22 fast-clearing) and performed rare variant association tests. These confirmed kelch as a major molecular marker for ART resistance (P = 6.03 × 10−6). This two-tier approach is powerful because pooled sequencing rapidly narrows down genome regions of interest, while targeted rare variant association testing within these regions can pinpoint the genetic basis of resistance. We show that our approach is robust to recurrent mutation and the generation of soft selective sweeps, which are predicted to be common in pathogen populations with large effective population sizes, and may confound more traditional gene mapping approaches.

The Plasmodium vivax merozoite surface protein 3β sequence reveals contrasting parasite populations in southern and northwestern Thailand.

BackgroundMalaria control efforts have a significant impact on the epidemiology and parasite population dynamics. In countries aiming for malaria elimination, malaria transmission may be restricted to limited transmission hot spots, where parasite populations may be isolated from each other and experience different selection forces. Here we aim to examine the Plasmodium vivax population divergence in geographically isolated transmission zones in Thailand.MethodologyWe employed the P. vivax merozoite surface protein 3β (PvMSP3β) as a molecular marker for characterizing P. vivax populations based on the extensive diversity of this gene in Southeast Asian parasite populations. To examine two parasite populations with different transmission levels in Thailand, we obtained 45 P. vivax isolates from Tak Province, northwestern Thailand, where the annual parasite incidence (API) was more than 2%, and 28 isolates from Yala and Narathiwat Provinces, southern Thailand, where the API was less than 0.02%. We sequenced the PvMSP3β gene and examined its genetic diversity and molecular evolution between the parasite populations.Principal FindingsOf 58 isolates containing single PvMSP3β alleles, 31 sequence types were identified. The overall haplotype diversity was 0.77±0.06 and nucleotide diversity 0.0877±0.0054. The northwestern vivax malaria population exhibited extensive haplotype diversity (HD) of PvMSP3β (HD = 1.0). In contrast, the southern parasite population displayed a single PvMSP3β allele (HD = 0), suggesting a clonal population expansion. This result revealed that the extent of allelic diversity in P. vivax populations in Thailand varies among endemic areas.ConclusionMalaria parasite populations in a given region may vary significantly in genetic diversity, which may be the result of control and influenced by the magnitude of malaria transmission intensity. This is an issue that should be taken into account for the implementation of P. vivax control measures such as drug policy and vaccine development.

Absence of putative artemisinin resistance mutations among Plasmodium falciparum in Sub-Saharan Africa: a molecular epidemiologic study.

Plasmodium falciparum parasites that are resistant to artemisinins have been detected in Southeast Asia. Resistance is associated with several polymorphisms in the parasite's K13-propeller gene. The molecular epidemiology of these artemisinin resistance genotypes in African parasite populations is unknown. We developed an assay to quantify rare polymorphisms in parasite populations that uses a pooled deep-sequencing approach to score allele frequencies, validated it by evaluating mixtures of laboratory parasite strains, and then used it to screen P. falciparum parasites from >1100 African infections collected since 2002 from 14 sites across sub-Saharan Africa. We found no mutations in African parasite populations that are associated with artemisinin resistance in Southeast Asian parasites. However, we observed 15 coding mutations, including 12 novel mutations, and limited allele sharing between parasite populations, consistent with a large reservoir of naturally occurring K13-propeller variation. Although polymorphisms associated with artemisinin resistance in P. falciparum in Southeast Asia are not prevalent in sub-Saharan Africa, numerous K13-propeller coding polymorphisms circulate in Africa. Although their distributions do not support a widespread selective sweep for an artemisinin-resistant phenotype, the impact of these mutations on artemisinin susceptibility is unknown and will require further characterization. Rapid, scalable molecular surveillance offers a useful adjunct in tracking and containing artemisinin resistance.

Evolutionary divergence of the swim bladder nematode Anguillicola crassus after colonization of a novel host, Anguilla anguilla

BackgroundAnguillicola crassus, a swim bladder nematode naturally parasitizing the Japanese eel, was introduced about 30 years ago from East Asia into Europe where it colonized almost all populations of the European eel. We conducted a common garden experiment under a reciprocal transfer design infecting both European and Japanese eels with populations of A. crassus from Germany, Poland and Taiwan. We tested, whether differences in infectivity, developmental dynamics and reproductive output between the European and Asian parasite populations occur while harboured in the specimens of native and colonized eel host, and if these differences are genetically based or are plastic responses to the new environment.ResultsUnder common garden conditions an evolutionary change in the both European parasite populations of A. crassus compared with their Taiwanese conspecifics was observed for infectivity and developmental dynamics, but not for reproductive output. When infecting the European eel, current European populations of the parasite were less infective and developed faster than their Taiwanese conspecifics. In the reciprocally infected Japanese eel the genetically induced differences between the parasite strains were less apparent than in the European eel but higher infectivity, faster development and higher larval mortality of the European parasite populations could be inferred.ConclusionsThe differences in infectivity and developmental dynamics between European and Taiwanese populations of A. crassus found in our study suggest rapid genetic divergence of this parasite after a successful host switch in Europe.

Plasmodium vivax populations revisited: mitochondrial genomes of temperate strains in Asia suggest ancient population expansion

BackgroundPlasmodium vivax is the most widely distributed human malaria parasite outside of Africa, and its range extends well into the temperate zones. Previous studies provided evidence for vivax population differentiation, but temperate vivax parasites were not well represented in these analyses. Here we address this deficit by using complete mitochondrial (mt) genome sequences to elucidate the broad genetic diversity and population structure of P. vivax from temperate regions in East and Southeast Asia.ResultsFrom the complete mtDNA sequences of 99 clinical samples collected in China, Myanmar and Korea, a total of 30 different haplotypes were identified from 26 polymorphic sites. Significant differentiation between different East and Southeast Asian parasite populations was observed except for the comparison between populations from Korea and southern China. Haplotype patterns and structure diversity analysis showed coexistence of two different groups in East Asia, which were genetically related to the Southeast Asian population and Myanmar population, respectively. The demographic history of P. vivax, examined using neutrality tests and mismatch distribution analyses, revealed population expansion events across the entire P. vivax range and the Myanmar population. Bayesian skyline analysis further supported the occurrence of ancient P. vivax population expansion.ConclusionsThis study provided further resolution of the population structure and evolution of P. vivax, especially in temperate/warm-temperate endemic areas of Asia. The results revealed divergence of the P. vivax populations in temperate regions of China and Korea from other populations. Multiple analyses confirmed ancient population expansion of this parasite. The extensive genetic diversity of the P. vivax populations is consistent with phenotypic plasticity of the parasites, which has implications for malaria control.

An introduced Asian parasite threatens northeastern Pacific estuarine ecosystems

The introduced Asian parasitic bopyrid isopod, Orthione griffenis, was first discovered on the Pacific coast of North America in Washington in 1988 and next in California in 1992. The range of Orthione presently extends from British Columbia to Baja California, where it infests at least two species of the native estuary mud shrimp, Upogebia. Intense Orthi- one infestations are associated with the apparent demise of many local populations of Upogebia pugettensis yet nonindigenous origins of Orthione in North America and thus the ecological significance of its impacts have remained in doubt. Six criteria reveal that Orthione is introduced to North America: its conspecificity with disjunct Asian populations, its earliest (1950s) collections in Asia, its late discovery among symbiotic species associated with Upogebia, its historical absence, and its appearance in North America coincident with extensive new ballast water traffic from Asia. Orthione is the first recognized bopyrid isopod invasion globally. Coexistence of U. pugettensis, which are ecosystem engineers, with its newly acquired parasite cannot be assumed. Orthione threatens eastern Pacific estuary ecosystems where Upogebia were previously abundant.