Mature Parasites Research Articles

Patent Macracanthorhynchus ingens Infection in a 17-Month-Old Child, Ohio.

Limited data exist on human Macracanthorhynchus infections. We report an asymptomatic 17-month-old who passed eggs and an adult Macracanthorhynchus ingens worm, indicating parasite maturation and reproduction. Macracanthorhynchus ingens may have a greater capacity to mature in humans versus Macracanthorhynchus hirudinaceus.

Plasmodium falciparum maturation across the intra-erythrocytic cycle shifts the soft glassy viscoelastic properties of red blood cells from a liquid-like towards a solid-like behavior

The mechanical properties of erythrocytes have been investigated by different techniques. However, there are few reports on how the viscoelasticity of these cells varies during malaria disease. Here, we quantitatively map the viscoelastic properties of Plasmodium falciparum-parasitized human erythrocytes. We apply new methodologies based on optical tweezers to measure the viscoelastic properties and defocusing microscopy to measure the erythrocyte height profile, the overall cell volume, and its form factor, a crucial parameter to convert the complex elastic constant into complex shear modulus. The storage and loss shear moduli are obtained for each stage of parasite maturation inside red blood cells, while the former increase, the latter decrease. Employing a soft glassy rheology model, we obtain the power-law exponent for the storage and loss shear moduli, characterizing the soft glassy features of red blood cells in each parasite maturation stage. Ring forms present a liquid-like behavior, with a slightly lower power-law exponent than healthy erythrocytes, whereas trophozoite and schizont stages exhibit increasingly solid-like behaviors. Finally, the surface elastic shear moduli, low-frequency surface viscosities, and shape recovery relaxation times all increase not only in a stage-dependent manner but also when compared to healthy red blood cells. Overall, the results call attention to the soft glassy characteristics of Plasmodium falciparum-parasitized erythrocyte membrane and may provide a basis for future studies to better understand malaria disease from a mechanobiological perspective.

Japanese Encephalitis and Associated Environmental Risk Factors in Eastern Uttar Pradesh: A time series analysis from 2001 to 2016

India and other Southeast Asian countries are severely affected by Japanese encephalitis (JE), one of the deadliest vector-borne disease threat to human health. Several epidemiological observations suggest climate variables play a role in providing a favorable environment for mosquito development and virus transmission. In this study, generalized additive models were used to determine the association of JE admissions and mortality with climate variables in Gorakhpur district, India, from 2001–2016. The model predicted that every 1 unit increase in mean (Tmean;°C), and minimum (Tmin;°C) temperature, rainfall (RF; mm) and relative humidity (RH; %) would on average increase the JE admissions by 22.23 %, 17.83 %, 0.66 %, and 5.22 % respectively and JE mortality by 13.27 %, 11.77 %, 0.94 %, and 3.27 % respectively Conversely, every unit decrease in solar radiation (Srad; MJ/m2/day) and wind speed (WS; Kmph) caused an increase in JE admission by 17% and 11.42% and in JE mortality by 9.37% and 4.88% respectively suggesting a protective effect at higher levels. The seasonal analysis shows that temperature was significantly associated with JE in pre-monsoon and post-monsoon while RF, RH, Srad, and WS are associated with the monsoon. Effect modification due to age and gender showed an equal risk for both genders and increased risk for adults above 15 years of age, however, males and age groups under 15 years outnumbered females and adults. Sensitivity analysis results to explore lag effects in climate variables showed that climate variables show the strongest association at lag 1 to 1.5 months with significant lag effect up tp lag 0-60 days. The exposure-response curve for climate variables showed a more or less linear relationship, with an increase in JE admissions and mortality after a certain threshold and decrease were reported at extreme levels of exposure. The study concludes that climate variables could influence the JE vector development and multiplication and parasite maturation and transmission in the Gorakhpur region whose indirect impact was noted for JE admission and mortality. In response to the changing climate, public health interventions, public awareness, and early warning systems would play an unprecedented role to compensate for future risk.

Live-Cell FRET Reveals that Malaria Nutrient Channel Proteins CLAG3 and RhopH2 Remain Associated throughout Their Tortuous Trafficking

Malaria parasites increase their host erythrocyte's permeability to various nutrients, fueling intracellular pathogen development and replication. The plasmodial surface anion channel (PSAC) mediates this uptake and is linked to the parasite-encoded RhopH complex, consisting of CLAG3, RhopH2, and RhopH3. While interactions between these subunits are well established, it is not clear whether they remain associated from their synthesis in developing merozoites through erythrocyte invasion and trafficking to the host membrane. Here, we explored protein-protein interactions between RhopH subunits using live-cell imaging and Förster resonance energy transfer (FRET) experiments. Using the green fluorescent protein (GFP) derivatives mCerulean and mVenus, we generated single- and double-tagged parasite lines for fluorescence measurements. While CLAG3-mCerulean served as an efficient FRET donor for RhopH2-mVenus within rhoptry organelles, mCerulean targeted to this organelle via a short signal sequence produced negligible FRET. Upon merozoite egress and reinvasion, these tagged RhopH subunits were deposited into the new host cell's parasitophorous vacuole; these proteins were then exported and trafficked to the erythrocyte membrane, where CLAG3 and RhopH2 remained fully associated. Fluorescence intensity measurements identified stoichiometric increases in exported RhopH protein when erythrocytes are infected with two parasites; whole-cell patch-clamp revealed a concomitant increase in PSAC functional copy number and a dose effect for RhopH contribution to ion and nutrient permeability. These studies establish live-cell FRET imaging in human malaria parasites, reveal that RhopH subunits traffic to their host membrane destination without dissociation, and suggest quantitative contribution to PSAC formation.IMPORTANCE Malaria parasites grow within circulating red blood cells and uptake nutrients through a pore on their host membrane. Here, we used gene editing to tag CLAG3 and RhopH2, two proteins linked to the nutrient pore, with fluorescent markers and tracked these proteins in living infected cells. After their synthesis in mature parasites, imaging showed that both proteins are packaged into membrane-bound rhoptries. When parasites ruptured their host cells and invaded new red blood cells, these proteins were detected within a vacuole around the parasite before they migrated and inserted in the surface membrane of the host cell. Using simultaneous labeling of CLAG3 and RhopH2, we determined that these proteins interact tightly during migration and after surface membrane insertion. Red blood cells infected with two parasites had twice the protein at their surface and a parallel increase in the number of nutrient pores. Our work suggests that these proteins directly facilitate parasite nutrient uptake from human plasma.

Relaxometric studies of erythrocyte suspensions infected by Plasmodium falciparum: a tool for staging infection and testing anti-malarial drugs.

Malaria is a global health problem with the most malignant form caused by Plasmodium falciparum (P. falciparum). Parasite maturation in red blood cells (RBCs) is accompanied by changes including the formation of paramagnetic hemozoin (HZ) nanocrystals, and increased metabolism and variation in membrane lipid composition. Herein, MR relaxometry (MRR) was applied to investigate water exchange across RBCs' membrane and HZ formation in parasitized RBCs. Transverse water protons relaxation rate constants (R2 = 1/T2 ) were measured for assessing HZ formation in P. falciparum-parasitized human RBCs. Moreover, water exchange lifetimes across the RBC membrane (τi ) were assessed by measuring longitudinal relaxation rate constants (R1 = 1/T1 ) at 21.5 MHz in the presence of a gadolinium complex dissolved in the suspension medium. τi increased after invasion of parasites (ring stage, mean τi / = 1.234 ± 0.022) and decreased during maturation to late trophozoite (mean τi / = 0.960 ± 0.075) and schizont stages (mean τi / = 1.019 ± 0.065). The HZ accumulation in advanced stages was revealed by T2 -shortening. The curves reporting R2 (1/T2 ) vs. magnetic field showed different slopes for non-parasitized RBCs (npRBCs) and parasitized RBCs (pRBCs), namely 0.003 ± 0.001 for npRBCs, 0.009 ± 0.002, 0.028 ± 0.004 and 0.055 ± 0.002 for pRBCs at ring-, early trophozoite-, and late trophozoite stage, respectively. Antimalarial molecules dihydroartemisinin and chloroquine elicited measurable changes in parasitized RBCs, namely dihydroartemisinin modified τi , whereas the interference of chloroquine with HZ formation was detectable by a significant T2 increase. MRR can be considered a useful tool for reporting on P. falciparum blood stages and for screening potential antimalarial molecules.

Molecular characterization of Plasmodium falciparum PHISTb proteins as potential targets of naturally-acquired immunity against malaria

Background: Plasmodium falciparum causes the deadliest form of malaria in humans. Upon infection, the host’s infected red blood cells (iRBCs) are remodelled by exported parasite proteins in order to provide a niche for parasite development and maturation. Methods: Here we analysed the role of three PHISTb proteins Pf3D7_0532400, Pf3D7_1401600, and Pf3D7_1102500 by expressing recombinant proteins and evaluated antibody responses against these proteins using immune sera from malaria-exposed individuals from Kenya and The Gambia in Africa. Results: Our findings show that children and adults from malaria-endemic regions recognized the three PHISTb proteins. Responses against the PHISTb proteins varied with malaria transmission intensity in three different geographical sites in Kenya (Siaya and Takaungu) and The Gambia (Sukuta). Antibody responses against PHISTb antigens Pf3D7_1102500 and Pf3D7_1401600 were higher in Sukuta, a low transmission region in the Gambia, as compared to Siaya, a high transmission region in western Kenya, unlike Pf3D7_0532400. Anti-PHIST responses show a negative correlation between antibody levels and malaria transmission intensity for two PHIST antigens, Pf3D7_1102500 and Pf3D7_1401600. However, we report a correlation in antibody responses between schizont extract and Pf3D7_0532400 (p=0.00582). Acquisition of anti-PHIST antibodies was correlated with exposure to malaria for PHISTb protein Pf3D7_0532400 (p=0.009) but not the other PHIST antigens Pf3D7_1102500 and Pf3D7_1401600 (p=0.507 and p=0.15, respectively, CI=95%). Children aged below 2 years had the lowest antibody levels, but the responses do not correlate with age differences. Conclusions: Collectively, these findings provide evidence of natural immunity against PHISTb antigens that varies with level of malaria exposure and underscore potential for these parasite antigens as possible serological markers to P. falciparum infection aimed at contributing to malaria control through vaccine development.

Inhibition of the NAD salvage pathway in schistosomes impairs metabolism, reproduction, and parasite survival.

NAD, a key co-enzyme required for cell metabolism, is synthesized via two pathways in most organisms. Since schistosomes apparently lack enzymes required for de novo NAD biosynthesis, we evaluated whether these parasites, which infect >200 million people worldwide, maintain NAD homeostasis via the NAD salvage biosynthetic pathway. We found that intracellular NAD levels decline in schistosomes treated with drugs that block production of nicotinamide or nicotinamide mononucleotide–known NAD precursors in the non-deamidating salvage pathway. Moreover, in vitro inhibition of the NAD salvage pathway in schistosomes impaired egg production, disrupted the outer membranes of both immature and mature parasites and caused loss of mobility and death. Inhibiting the NAD salvage pathway in schistosome-infected mice significantly decreased NAD levels in adult parasites, which correlated with reduced egg production, fewer liver granulomas and parasite death. Thus, schistosomes, unlike their mammalian hosts, appear limited to one metabolic pathway to maintain NAD-dependent metabolic processes.

Distribution of fish apophalosis in the river basins of Sumy region

The causative agent of apophalosis (rosicotremosis) is the larva of Apofalus donicus. It is localized in the skin, scales, perch fins, rust, walleye, and some carp fishes that live in rivers flowing into the Black Sea, in the Tisza River, and in the Azov Sea estuaries. Mature trematode parasites in the small intestine the dogs, cats, foxes, some fish-eating birds (common buzzards, quarks, white storks) and humans. The purpose of the research was to conduct research to determine the incidence of fish for trematodoses and to develop measures to ensure the regimes of fish disinfection when they are detected. The research was conducted on the basis of the department of veterinary medicine, microbiology, zooghygiene and safety and quality of animal products of the Faculty of Veterinary Medicine of Sumy National Agrarian University. During the period 2018–2019, fish were caught from the rivers of the Dnieper Basin: Sula, Psla and Vorskla. The main species of fish studied are: carp – Cyprinus carpio (L.), perch – Perca fluviatilis (L.); common ruff (Gymnocephalus cernua), walleye (Sander iucioperca). A total of 321 fish were examined. Parasitological examination of the fish was carried out by the method of complete parasitological dissection, which makes it possible to conduct quantitative and qualitative accounting of all helminths affected by the fish. The autopsy of the fish was performed according to the conventional method. The incision was made from the anal fin up and forward to the gill lid just above the base of the pectoral fin. As a result of the fish studies, the larvae of trematodes were identified, which by structure were classified as Apofalus donicus. The larvae were alive, localized under the skin and moved. The overall invasion intensity for all species of fish studied was 3.64%. In the next phase of the study we conducted experiments to determine the resistance of the larvae to different modes of freezing, salinization and heat treatment. effective for decontamination of fish meat with the presence of metacercariae are: boiling at 80 °C, 15 minutes, at 100 °C, 10 minutes; freezing -18 °С, 7 days; salting of 12 % tuzluk 1 month; frying – from 15 minutes; action of microwaves 480 W, 8 minutes, 760 W, 4 minutes. The prevalence of apophalosis among fish of the Dnieper River basin (Sula, Psel, Vorskla) in Sumy region was established. The overall invasion intensity for all species of fish studied was 3.64 %. Effective for the disinfection of fish meat with the presence of Apofalus donicus metacercariae are: boiling at 80 °C, 15 minutes, at 100 °C, 10 minutes; freezing -18 °С, 7 days; salting of 12 % tuzluk 1 month; frying – from 15 minutes; action of microwaves 480 W, 8 minutes, 760 W, 4 minutes. In the future, it is planned to conduct research to determine the spread of apophalosis among cats, dogs and fish birds.

Essential function of the alveolin network in the subpellicular microtubules and conoid assembly in Toxoplasma gondii.

The coccidian subgroup of Apicomplexa possesses an apical complex harboring a conoid, made of unique tubulin polymer fibers. This enigmatic organelle extrudes in extracellular invasive parasites and is associated to the apical polar ring (APR). The APR serves as microtubule-organizing center for the 22 subpellicular microtubules (SPMTs) that are linked to a patchwork of flattened vesicles, via an intricate network composed of alveolins. Here, we capitalize on ultrastructure expansion microscopy (U-ExM) to localize the Toxoplasma gondii Apical Cap protein 9 (AC9) and its partner AC10, identified by BioID, to the alveolin network and intercalated between the SPMTs. Parasites conditionally depleted in AC9 or AC10 replicate normally but are defective in microneme secretion and fail to invade and egress from infected cells. Electron microscopy revealed that the mature parasite mutants are conoidless, while U-ExM highlighted the disorganization of the SPMTs which likely results in the catastrophic loss of APR and conoid.

Ensemble modeling highlights importance of understanding parasite-host behavior in preclinical antimalarial drug development

Emerging drug resistance and high-attrition rates in early and late stage drug development necessitate accelerated development of antimalarial compounds. However, systematic and meaningful translation of drug efficacy and host-parasite dynamics between preclinical testing stages is missing. We developed an ensemble of mathematical within-host parasite growth and antimalarial action models, fitted to extensive data from four antimalarials with different modes of action, to assess host-parasite interactions in two preclinical drug testing systems of murine parasite P. berghei in mice, and human parasite P. falciparum in immune-deficient mice. We find properties of the host-parasite system, namely resource availability, parasite maturation and virulence, drive P. berghei dynamics and drug efficacy, whereas experimental constraints primarily influence P. falciparum infection and drug efficacy. Furthermore, uninvestigated parasite behavior such as dormancy influences parasite recrudescence following non-curative treatment and requires further investigation. Taken together, host-parasite interactions should be considered for meaningful translation of pharmacodynamic properties between murine systems and for predicting human efficacious treatment.

Parasites and Maturation Rate as Indicators of Population Structure in the Beaked Redfish Sebastes mentella (Sebastidae)

The direction of phylogenesis of beaked redfish Sebastes mentella is considered based on a comparative analysis of the parasite fauna and maturation rate of individuals in the pelagic and near-bottom groups in the North Atlantic and the Norwegian-Barents sea populations. The individuals of the beaked redfish inhabiting the pelagic and bathyal zones of the Norwegian Sea’s northern part are characterized by a significantly higher similarity of the parasite fauna and lesser differences in the length and maturation age than that living in the pelagic and bathyal zones of the North Atlantic seas. This may indicate that the pelagic group of beaked redfish in the Norwegian Sea is at an earlier stage of formation than the pelagic group in the Irminger and Labrador seas, and the Norwegian-Barents sea population is phylogenetically younger than the North Atlantic one. The occurrence of mass migration of beaked redfish individuals from the pelagic zone of the Irminger Sea to the Norwegian Sea indicates a probable source of origin of its Norwegian-Barents sea population.

Single-cell RNA sequencing of Trypanosoma brucei from tsetse salivary glands unveils metacyclogenesis and identifies potential transmission blocking antigens

Tsetse-transmitted African trypanosomes must develop into mammalian-infectious metacyclic cells in the fly's salivary glands (SGs) before transmission to a new host. The molecular mechanisms that underlie this developmental process, known as metacyclogenesis, are poorly understood. Blocking the few metacyclic parasites deposited in saliva from further development in the mammal could prevent disease. To obtain an in-depth perspective of metacyclogenesis, we performed single-cell RNA sequencing (scRNA-seq) from a pool of 2,045 parasites collected from infected tsetse SGs. Our data revealed three major cell clusters that represent the epimastigote, and pre- and mature metacyclic trypanosome developmental stages. Individual cell level data also confirm that the metacyclic pool is diverse, and that each parasite expresses only one of the unique metacyclic variant surface glycoprotein (mVSG) coat protein transcripts identified. Further clustering of cells revealed a dynamic transcriptomic and metabolic landscape reflective of a developmental program leading to infectious metacyclic forms preadapted to survive in the mammalian host environment. We describe the expression profile of proteins that regulate gene expression and that potentially play a role in metacyclogenesis. We also report on a family of nonvariant surface proteins (Fam10) and demonstrate surface localization of one member (named SGM1.7) on mature metacyclic parasites. Vaccination of mice with recombinant SGM1.7 reduced parasitemia early in the infection. Future studies are warranted to investigate Fam10 family proteins as potential trypanosome transmission blocking vaccine antigens. Our experimental approach is translationally relevant for developing strategies to prevent other insect saliva-transmitted parasites from infecting and causing disease in mammalian hosts.

Methods to Investigate the Deformability of RBC During Malaria.

Despite a 30% decline in mortality since 2000, malaria still affected 219 million subjects and caused 435,000 deaths in 2017. Red blood cells (RBC) host Plasmodium parasites that cause malaria, of which Plasmodium falciparum is the most pathogenic. The deformability of RBC is markedly modified by invasion and development of P. falciparum. Surface membrane area is potentially impacted by parasite entry and development, the cytoskeleton is modified by parasite proteins and cytosol viscosity is altered by parasite metabolism. RBC hosting mature parasites (second half of the asexual erythrocytic cycle) are abnormally stiff but the main reason for their absence from the circulation is their adherence to endothelial cells, mediated by parasite proteins exposed at the infected-RBC surface. By contrast, the circulation of non-adherent rings and gametocytes, depends predominantly on deformability. Altered deformability of rings and of uninfected-RBC altered by malaria infection is an important determinant of malaria pathogenesis. It also impacts the response to antimalarial therapy. Unlike conventional antimalarials that target mature stages, currently recommended first-line artemisinin derivatives and the emerging spiroindolones act on circulating rings. Methods to investigate the deformability of RBC are therefore critical to understand the clearance of infected- and uninfected-RBC in malaria. Herein, we review the main methods to assess the deformability of P. falciparum infected-RBC, and their contribution to the understanding of how P. falciparum infection causes disease, how the parasite is transmitted and how antimalarial drugs induce parasite clearance.

Myxobolus spp. (Cnidaria: Myxozoa) in introduced yellow perch Perca flavescens (Mitchill, 1814)

We surveyed introduced yellow perch Perca flavescens (Mitchill, 1814) from the Willamette River, OR, USA, to determine if these fish have co-introduced myxosporean parasites. Mature parasite myxospores were observed in brains of 3/19 fish, and were morphologically and molecularly consistent with Myxobolus neurophilus (Guilford 1963), a parasite known from yellow perch in their native range. We identified another Myxobolus species from the gill filaments of 1/22 fish. The spores from the gill filaments were oval-shaped, 11.7 (10.7-12.3) μm long × 8.6 (7.7-9.0) μm wide × 5.2 (4.6-5.6) μm thick, with two oval-shaped polar capsules 5.7 (5.1-6.5) μm×2.7 (2.4-3.2) μm, each containing a polar tubule with 8-9 turns. Small-subunit ribosomal DNA sequences from each of four plasmodia were identical, and 4.0% different (over 1800 nucleotides) from the closest known myxosporeans. Interestingly, these sequences had overlapping peaks in their chromatograms, which suggested that DNA from multiple species was present. Hence, we isolated and sequenced three individual myxospores and found that they too had mixed chromatograms, which indicated presence of at least two sequence types of small-subunit ribosomal DNA in each spore (GenBank accession MK592012, MK592013), a rare character among described myxosporeans. The spore morphology, morphometry, tissue tropism, and DNA sequence supported a diagnosis of a novel species, Myxobolus doubleae n. sp. This parasite is unknown from yellow perch in its native range, despite extensive historical surveys, which suggests that introduced yellow perch might have acquired an endemic Myxobolus species via spillback from another fish host.

A new genus and species of parasitic isopod (Bopyroidea: Entoniscidae) infesting pinnotherid crabs (Brachyura: Pinnotheridae) on the Atlantic coast of the USA, with notes on the life cycle of entoniscids

AbstractPinnixion sexdecennia n. gen., n. sp. is described from three pinnotherid hosts: Austinixa gorei (Manning & Felder, 1989), Pinnixa chaetopterana Stimpson, 1860, and Zaops ostreus (Say, 1817). Females of the new species are distinguished from all other entoniscids by possessing two transverse lobes on oostegite 1 and fused oostegites 2–5; males have a pair of lateral expansions on each of the first two pleomeres. The epicaridium larvae of the new species also have a unique elongated, cylindrical terminal process on the pereopod 6 dactylus. Pinnixion sexdecennian. gen., n. sp. exhibited highest prevalence (> 40%) in North Carolina and Florida. Examination of live specimens of P. chaetopterana infested with mature female isopods showed that larvae are liberated via a pore produced in the wall of the host’s branchial chamber. Mature females of P. sexdecennian. gen., n. sp. occupied nearly all available space within the host crab’s hemocoel. Female P. chaetopterana had higher prevalence than males, and intensities of infestations were also greater in females. Evidence from all locations showed that mature parasites do not inhibit breeding or molting in the host. Our report is one of the few studies on the ecology and biology of entoniscids, a cryptic but ecologically important group of parasitic isopods. We also resolve long-standing nomenclatural issues involving Entione Kossmann, 1881 and Entioninae Codreanu, Codreanu & Pike, 1960, the subfamily to which the new genus and species belongs, by synonymizing Grapsion Giard & Bonnier, 1886 with Entione.

Profiling Transcriptional Regulation and Functional Roles of Schistosoma mansoni c-Jun N-Terminal Kinase.

Mitogen-activated protein kinases (MAPKs) play a regulatory role and influence various biological activities, such as cell proliferation, differentiation, and survival. Our group has demonstrated through functional studies that Schistosoma mansoni c-Jun N-terminal kinase (SmJNK) MAPK is involved in the parasite’s development, reproduction, and survival. SmJNK can, therefore, be considered a potential target for the development of new drugs. Considering the importance of SmJNK in S. mansoni maturation, we aimed at understanding of SmJNK regulated signaling pathways in the parasite, correlating expression data with S. mansoni development. To better understand the role of SmJNK in S. mansoni intravertebrate host life stages, RNA interference knockdown was performed in adult worms and in schistosomula larval stage. SmJNK knocked-down in adult worms showed a decrease in oviposition and no significant alteration in their movement. RNASeq libraries of SmJNK knockdown schistosomula were sequenced. A total of 495 differentially expressed genes were observed in the SmJNK knockdown parasites, of which 373 were down-regulated and 122 up-regulated. Among the down-regulated genes, we found transcripts related to protein folding, purine nucleotide metabolism, the structural composition of ribosomes and cytoskeleton. Genes coding for proteins that bind to nucleic acids and proteins involved in the phagosome and spliceosome pathways were enriched. Additionally, we found that SmJNK and Smp38 MAPK signaling pathways converge regulating the expression of a large set of genes. C. elegans orthologous genes were enriched for genes related to sterility and oocyte maturation, corroborating the observed phenotype alteration. This work allowed an in-depth analysis of the SmJNK signaling pathway, elucidating gene targets of regulation and functional roles of this critical kinase for parasite maturation.

Mechanisms Involved in the Persistence of Babesia canis Infection in Dogs.

Dogs that are infected with Babesia canis parasites usually show severe clinical signs, yet often very few parasites are detectable in the blood circulation. The results showed that large numbers of B. canis-infected red blood cells accumulate in the microvasculature of infected subjects. The initial process leading to the attachment of infected erythrocytes to the endothelial cells of small capillaries (sequestration) appears to involve the interaction of parasite molecules at the erythrocyte surface with ligands on the endothelial cells. Since parasites continue to develop in the sequestered erythrocyte, it would be expected that the infected erythrocyte is destroyed when the mature parasites escape the host cell, which would make it hard to explain accumulation of infected erythrocytes at the initial site of attachment. Apparently, additional processes are triggered that lead to consolidation of parasite sequestration. One possible explanation is that after initial attachment of an infected erythrocyte to the wall of a blood capillary, the coagulation system is involved in the trapping of infected and uninfected erythrocytes. The data further suggest that newly formed parasites subsequently infect normal red blood cells that are also trapped in the capillary, which finally leads to capillaries that appear to be loaded with infected erythrocytes.

Gut Microbiota Composition Is Associated with Severity of Disease in Patients with <i>P. Falciparum</i> Malaria

Background: In severe Plasmodium falciparum malaria, mature blood-stage parasites sequester in the intestinal microvasculature and compromise gut barrier integrity, with subsequent translocation of Gram-negative bacteria and bacterial products into the circulation. The role of the gut microbiota in the pathogenesis of life-threatening complications of severe malaria, such as acidosis or concomitant sepsis, is unknown. Methods: A prospective observational study was conducted to profile the gut microbiota composition of adult patients with severe and uncomplicated falciparum malaria and healthy volunteers in Bangladesh, by sequencing the V4 region of the 16S rRNA gene PCR-amplified from faecal DNA. Plasma markers of P. falciparum parasite biomass and gut barrier integrity were measured to determine the relationship between the gut microbiota and severity of disease. Findings: Patients with high parasite biomass had reduced gut integrity and increased levels of gut-derived D-lactate in their blood. The gut microbiota of patients with severe malaria was significantly enriched with potentially pathogenic Enteroccocus and Escherichia/Shigella species. Plasma D-lactate levels were associated with the abundance of known lactate-producing species in the gut microbiota. Interpretation: This first report of the gut microbiota in patients with severe malaria shows a pathogenic host-parasite-microbe interaction, and highlights the gut microbiota as a target for adjunctive therapies. Trial Registration: The trial was registered at ClinicalTrials.gov [NCT02451904]. Funding Statement: This work was supported by the Wellcome Trust [106698/B14/Z], a Nuffield Department of Medicine Prize Scholarship (SJL), the Netherlands Organization for Scientifc Research (VIDI-grant to WJW and Spinoza Award and SIAM Gravitation - grant 024.002.002 to WMdV). Declaration of Interests: The authors declare no competing interests. Ethical Approval Statement: This was a sub-study of an observational study protocol that was reviewed and approved by the local ethics review board, the Chittagong Medical College Ethics Committee, Bangladesh, and the Oxford Tropical Research Ethics Committee (OxTREC), in the United Kingdom.

Development of Onchocerca volvulus in humanized NSG mice and detection of parasite biomarkers in urine and serum.

BackgroundThe study of Onchocerca volvulus has been limited by its host range, with only humans and non-human primates shown to be susceptible to the full life cycle infection. Small animal models that support the development of adult parasites have not been identified.Methodology/Principal findingsWe hypothesized that highly immunodeficient NSG mice would support the survival and maturation of O. volvulus and alteration of the host microenvironment through the addition of various human cells and tissues would further enhance the level of parasite maturation. NSG mice were humanized with: (1) umbilical cord derived CD34+ stem cells, (2) fetal derived liver, thymus and CD34+ stem cells or (3) primary human skeletal muscle cells. NSG and humanized NSG mice were infected with 100 O. volvulus infective larvae (L3) for 4 to 12 weeks. When necropsies of infected animals were performed, it was observed that parasites survived and developed throughout the infection time course. In each of the different humanized mouse models, worms matured from L3 to advanced fourth stage larvae, with both male and female organ development. In addition, worms increased in length by up to 4-fold. Serum and urine, collected from humanized mice for identification of potential biomarkers of infection, allowed for the identification of 10 O. volvulus-derived proteins found specifically in either the urine or the serum of the humanized O. volvulus-infected NSG mice.Conclusions/SignificanceThe newly identified mouse models for onchocerciasis will enable the development of O. volvulus specific biomarkers, screening for new therapeutic approaches and potentially studying the human immune response to infection with O. volvulus.

Retraction Note to: Occurrence, spread and control measures of Bothriocephalus acheilognathi (Bothriocephalidae: Cestoda).

[This retracts the article DOI: 10.1007/s12639-014-0568-6.].