ResearchPad - cestodes https://www.researchpad.co Default RSS Feed en-us © 2020 Newgen KnowledgeWorks <![CDATA[Genetic diversity of <i>Echinococcus multilocularis</i> and <i>Echinococcus granulosus sensu lato</i> in Kyrgyzstan: The A2 haplotype of <i>E</i>. <i>multilocularis</i> is the predominant variant infecting humans]]> https://www.researchpad.co/article/elastic_article_13871 Analysis of the genetic variability in Echinococcus species from different endemic countries have contributed to the knowledge in the taxonomy and phylogeography of these parasites. The most important species of this genus, Echinococcus granulosus sensu lato and Echinococcus multilocularis, co-exist in Kyrgyzstan causing serious public health issues. E. granulosus s.l. causes cystic echinococcosis and E. multilocularis is the causative agent of alveolar echinococcosis. The most relevant finding of our study is the identification of the cob/nad2/cox1 A2 haplotype of E. multilocularis as the most commonly found in humans and dogs. However, it remains unknown if this variant of E. multilocularis, based on genetic differences in mitochondrial genes, presents differences in virulence which could have contributed to the emergence of alveolar echinococcosis in Kyrgyzstan. The results also show a number of non-previously described genetic variants of E. multilocularis and E. granulosus s.s.

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<![CDATA[New insights of the local immune response against both fertile and infertile hydatid cysts]]> https://www.researchpad.co/article/5c5b5259d5eed0c4842bc6ca

Background

Cystic echinococcosis is caused by the metacestode of the zoonotic flatworm Echinococcus granulosus. Within the viscera of the intermediate host, the metacestode grows as a unilocular cyst known as hydatid cyst. This cyst is comprised of two layers of parasite origin: germinal and laminated layers, and one of host origin: the adventitial layer, that encapsulates the parasite. This adventitial layer is composed of collagen fibers, epithelioid cells, eosinophils and lymphocytes. To establish itself inside the host, the germinal layer produces the laminated layer, and to continue its life cycle, generates protoscoleces. Some cysts are unable to produce protoscoleces, and are defined as infertile cysts. The molecular mechanisms involved in cyst fertility are not clear, however, the host immune response could play a crucial role.

Methodology/Principal findings

We collected hydatid cysts from both liver and lungs of slaughtered cattle, and histological sections of fertile, infertile and small hydatid cysts were stained with haematoxylin-eosin. A common feature observed in infertile cysts was the disorganization of the laminated layer by the infiltration of host immune cells. These infiltrating cells eventually destroy parts of laminated layer. Immunohistochemical analysis of both parasite and host antigens, identify these cells as cattle macrophages and are present inside the cysts associated to germinal layer.

Conclusions/Significance

This is the first report that indicates to cell from immune system present in adventitial layer of infertile bovine hydatid cysts could disrupt the laminated layer, infiltrating and probably causing the infertility of cyst.

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<![CDATA[Extracellular vesicles from Echinococcus granulosus larval stage: Isolation, characterization and uptake by dendritic cells]]> https://www.researchpad.co/article/5c3d00e7d5eed0c4840368ed

The secretion of extracellular vesicles (EVs) in helminth parasites is a constitutive mechanism that promotes survival by improving their colonization and adaptation in the host tissue. In the present study, we analyzed the production of EVs from supernatants of cultures of Echinococcus granulosus protoscoleces and metacestodes and their interaction with dendritic cells, which have the ability to efficiently uptake and process microbial antigens, activating T lymphocytes. To experimentally increase the release of EVs, we used loperamide, a calcium channel blocker that increases the cytosolic calcium level in protoscoleces and EV secretion. An exosome-like enriched EV fraction isolated from the parasite culture medium was characterized by dynamic light scattering, transmission electron microscopy, proteomic analysis and immunoblot. This allowed identifying many proteins including: small EV markers such as TSG101, SDCBP, ALIX, tetraspanins and 14-3-3 proteins; proteins involved in vesicle-related transport; orthologs of mammalian proteins involved in the immune response, such as basigin, Bp29 and maspardin; and parasite antigens such as antigen 5, P29 and endophilin-1, which are of special interest due to their role in the parasite-host relationship. Finally, studies on the EVs-host cell interaction demonstrated that E. granulosus exosome-like vesicles were internalized by murine dendritic cells, inducing their maturation with increase of CD86 and with a slight down-regulation in the expression of MHCII molecules. These data suggest that E. granulosus EVs could interfere with the antigen presentation pathway of murine dendritic cells inducing immunoregulation in the host. Further studies are needed to better understand the role of these vesicles in parasite survival and as diagnostic markers and new vaccines.

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<![CDATA[The ectodomains of the lymphocyte scavenger receptors CD5 and CD6 interact with tegumental antigens from Echinococcus granulosus sensu lato and protect mice against secondary cystic echinococcosis]]> https://www.researchpad.co/article/5c0ae451d5eed0c4845895e9

Background

Scavenger Receptors (SRs) from the host’s innate immune system are known to bind multiple ligands to promote the removal of non-self or altered-self targets. CD5 and CD6 are two highly homologous class I SRs mainly expressed on all T cells and the B1a cell subset, and involved in the fine tuning of activation and differentiation signals delivered by the antigen-specific receptors (TCR and BCR, respectively), to which they physically associate. Additionally, CD5 and CD6 have been shown to interact with and sense the presence of conserved pathogen-associated structures from bacteria, fungi and/or viruses.

Methodology/Principal findings

We report herein the interaction of CD5 and CD6 lymphocyte surface receptors with Echinococcus granulosus sensu lato (s.l.). Binding studies show that both soluble and membrane-bound forms of CD5 and CD6 bind to intact viable protoscoleces from E. granulosus s.l. through recognition of metaperiodate-resistant tegumental components. Proteomic analyses allowed identification of thioredoxin peroxidase for CD5, and peptidyl-prolyl cis-trans isomerase (cyclophilin) and endophilin B1 (antigen P-29) for CD6, as their potential interactors. Further in vitro assays demonstrate that membrane-bound or soluble CD5 and CD6 forms differentially modulate the pro- and anti-inflammatory cytokine release induced following peritoneal cells exposure to E. granulosus s.l. tegumental components. Importantly, prophylactic infusion of soluble CD5 or CD6 significantly ameliorated the infection outcome in the mouse model of secondary cystic echinococcosis.

Conclusions/Significance

Taken together, the results expand the pathogen binding properties of CD5 and CD6 and provide novel evidence for their therapeutic potential in human cystic echinococcosis.

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<![CDATA[Proteomic Analysis of Excretory-Secretory Products of Mesocestoides corti Metacestodes Reveals Potential Suppressors of Dendritic Cell Functions]]> https://www.researchpad.co/article/5989da24ab0ee8fa60b80016

Accumulating evidences have assigned a central role to parasite-derived proteins in immunomodulation. Here, we report on the proteomic identification and characterization of immunomodulatory excretory-secretory (ES) products from the metacestode larva (tetrathyridium) of the tapeworm Mesocestoides corti (syn. M. vogae). We demonstrate that ES products but not larval homogenates inhibit the stimuli-driven release of the pro-inflammatory, Th1-inducing cytokine IL-12p70 by murine bone marrow-derived dendritic cells (BMDCs). Within the ES fraction, we biochemically narrowed down the immunosuppressive activity to glycoproteins since active components were lipid-free, but sensitive to heat- and carbohydrate-treatment. Finally, using bioassay-guided chromatographic analyses assisted by comparative proteomics of active and inactive fractions of the ES products, we defined a comprehensive list of candidate proteins released by M. corti tetrathyridia as potential suppressors of DC functions. Our study provides a comprehensive library of somatic and ES products and highlight some candidate parasite factors that might drive the subversion of DC functions to facilitate the persistence of M. corti tetrathyridia in their hosts.

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<![CDATA[Development of a movement-based in vitro screening assay for the identification of new anti-cestodal compounds]]> https://www.researchpad.co/article/5989db5cab0ee8fa60be01c4

Intestinal cestodes are infecting millions of people and livestock worldwide, but treatment is mainly based on one drug: praziquantel. The identification of new anti-cestodal compounds is hampered by the lack of suitable screening assays. It is difficult, or even impossible, to evaluate drugs against adult cestodes in vitro due to the fact that these parasites cannot be cultured in microwell plates, and adult and larval stages in most cases represent different organisms in terms of size, morphology, and metabolic requirements. We here present an in vitro-drug screening assay based on Echinococcus multilocularis protoscoleces, which represent precursors of the scolex (hence the anterior part) of the adult tapeworm. This movement-based assay can serve as a model for an adult cestode screen. Protoscoleces are produced in large numbers in Mongolian gerbils and mice, their movement is measured and quantified by image analysis, and active compounds are directly assessed in terms of morphological effects. The use of the 384-well format minimizes the amount of parasites and compounds needed and allows rapid screening of a large number of chemicals. Standard drugs showed the expected dose-dependent effect on movement and morphology of the protoscoleces. Interestingly, praziquantel inhibited movement only partially within 12 h of treatment (at concentrations as high as 100 ppm) and did thus not act parasiticidal, which was also confirmed by trypan blue staining. Enantiomers of praziquantel showed a clear difference in their minimal inhibitory concentration in the motility assay and (R)-(-)-praziquantel was 185 times more active than (S)-(-)-praziquantel. One compound named MMV665807, which was obtained from the open access MMV (Medicines for Malaria Venture) Malaria box, strongly impaired motility and viability of protoscoleces. Corresponding morphological alterations were visualized by scanning electron microscopy, and demonstrated that this compound exhibits a mode of action clearly distinct from praziquantel. Thus, MMV665807 represents an interesting lead for further evaluation.

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<![CDATA[When Parasites Are Good for Health: Cestode Parasitism Increases Resistance to Arsenic in Brine Shrimps]]> https://www.researchpad.co/article/5989db46ab0ee8fa60bd8831

Parasites and pollutants can both affect any living organism, and their interactions can be very important. To date, repeated studies have found that parasites and heavy metals or metalloids both have important negative effects on the health of animals, often in a synergistic manner. Here, we show for the first time that parasites can increase host resistance to metalloid arsenic, focusing on a clonal population of brine shrimp from the contaminated Odiel and Tinto estuary in SW Spain. We studied the effect of cestodes on the response of Artemia to arsenic (acute toxicity tests, 24h LC50) and found that infection consistently reduced mortality across a range of arsenic concentrations. An increase from 25°C to 29°C, simulating the change in mean temperature expected under climate change, increased arsenic toxicity, but the benefits of infection persisted. Infected individuals showed higher levels of catalase and glutathione reductase activity, antioxidant enzymes with a very important role in the protection against oxidative stress. Levels of TBARS were unaffected by parasites, suggesting that infection is not associated with oxidative damage. Moreover, infected Artemia had a higher number of carotenoid-rich lipid droplets which may also protect the host through the “survival of the fattest” principle and the antioxidant potential of carotenoids. This study illustrates the need to consider the multi-stress context (contaminants and temperature increase) in which host-parasite interactions occur.

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<![CDATA[Trace element and metal sequestration in vitellaria and sclerites, and reactive oxygen intermediates in a freshwater monogenean, Paradiplozoon ichthyoxanthon]]> https://www.researchpad.co/article/5989db5aab0ee8fa60bdf780

Exposure to metals and other trace elements negatively affects infection dynamics of monogeneans, including diplozoids, but, physiological mechanisms linked to exposure have yet to be documented. In this study sequestration of trace elements and reactive oxygen intermediate production in the monogenean, Paradiplozoon ichthyoxanthon, was demonstrated. During dissection of host fish, Labeobarbus aeneus, the gills were excised and assessed for P. ichthyoxanthon, which were removed and frozen for fluorescence microscopy or fixed for transmission electron microscopy. Trace elements were sequestered in the vitellaria and sclerites in P. ichthyoxanthon, and the presence of reactive oxygen intermediates was observed predominantly in the tegument of the parasite. Trace elements and metals identified and ranked according to weight percentages (wt%) in the vitellaria were Cu > C > Au > O > Cr > Fe > Si while for the sclerites C > Cu > O > Au > Fe > Cr > Si were identified. For most element detected, readings were higher in the vitellaria than the sclerites, except for C and O which were higher in sclerites. Specifically for metals, all levels detected in the vitellaria were greater than in sclerites. Based on the proportion of trace elements present in the vitellaria and sclerites it appears that most trace elements including metals were sequestered in the vitellaria. The results of reactive oxygen intermediate production in the tegument of the parasite suggests either trace element accumulation takes place across the tegument or results from the action of the host’s immune response on the parasite. The results serve as the first demonstration of trace element sequestration and reactive oxygen intermediates in a freshwater monogenean parasite.

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<![CDATA[Functional Role of Native and Invasive Filter-Feeders, and the Effect of Parasites: Learning from Hypersaline Ecosystems]]> https://www.researchpad.co/article/5989d9f3ab0ee8fa60b6f5ed

Filter-feeding organisms are often keystone species with a major influence on the dynamics of aquatic ecosystems. Studies of filtering rates in such taxa are therefore vital in order to understand ecosystem functioning and the impact of natural and anthropogenic stressors such as parasites, climate warming and invasive species. Brine shrimps Artemia spp. are the dominant grazers in hypersaline systems and are a good example of such keystone taxa. Hypersaline ecosystems are relatively simplified environments compared with much more complex freshwater and marine ecosystems, making them suitable model systems to address these questions. The aim of this study was to compare feeding rates at different salinities and temperatures between clonal A. parthenogenetica (native to Eurasia and Africa) and the invasive American brine shrimp A. franciscana, which is excluding native Artemia from many localities. We considered how differences observed in laboratory experiments upscale at the ecosystem level across both spatial and temporal scales (as indicated by chlorophyll-a concentration and turbidity). In laboratory experiments, feeding rates increased at higher temperatures and salinities in both Artemia species and sexes, whilst A. franciscana consistently fed at higher rates. A field study of temporal dynamics revealed significantly higher concentrations of chlorophyll-a in sites occupied by A. parthenogenetica, supporting our experimental findings. Artemia parthenogenetica density and biomass were negatively correlated with chlorophyll-a concentration at the spatial scale. We also tested the effect of cestode parasites, which are highly prevalent in native Artemia but much rarer in the invasive species. The cestodes Flamingolepis liguloides and Anomotaenia tringae decreased feeding rates in native Artemia, whilst Confluaria podicipina had no significant effect. Total parasite prevalence was positively correlated with turbidity. Overall, parasites are likely to reduce feeding rates in the field, and their negative impact on host fecundity is likely to exacerbate the difference between grazing rates of native and alien Artemia populations at the ecosystem level. The results of this study provide evidence for the first time that the replacement of native Artemia by A. franciscana may have major consequences for the functioning of hypersaline ecosystems. The strong effect of parasites on feeding rate underlines the importance of taking parasites into account in order to improve our understanding of the functioning of aquatic ecosystems.

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<![CDATA[Eggs as a Suitable Tool for Species Diagnosis of Causative Agents of Human Diphyllobothriosis (Cestoda)]]> https://www.researchpad.co/article/5989db0aab0ee8fa60bc9dc2

Background

Tapeworms of the order Diphyllobothriidea are parasites of tetrapods and several species may infect man and cause neglected human disease called diphyllobothriosis. Identification of human-infecting diphyllobothriid cestodes is difficult because of their morphological uniformity, which concerns also their eggs in stool samples.

Methods

In the present study, we analysed by far the largest dataset of more than 2,000 eggs of 8 species of diphyllobothriid cestodes that may infect humans, including the most frequent human parasites Diphyllobothrium latum, D. nihonkaiense and Adenocephalus pacificus (syn. Diphyllobothrium pacificum). Size (length, width and length/width ratio) and the surface of the egg shell from naturally and experimentally infected hosts were studied using light and scanning electron microscopy.

Results

A high degree of intraspecific and host-related size variability has been detected, but combination of morphometrical and ultrastructural data made it possible to distinguish all of the studied species, including otherwise quite similar eggs of the 3 most common species infecting man, i.e. D. latum, D. nihonkaiense and D. dendriticum. The surface of all marine species is covered by numerous deep pits with species-specific density, whereas the surface of freshwater species is smooth or with isolated shallow hollows or wrinkles.

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