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Parasitology Nov 2022is the aetiological agent of cysticercosis, a zoonosis that causes severe health and economic losses across Latin America, Africa and Asia. The most serious...
is the aetiological agent of cysticercosis, a zoonosis that causes severe health and economic losses across Latin America, Africa and Asia. The most serious manifestation of the disease is neurocysticercosis, which occurs when the larval stage (cysticercus) establishes in the central nervous system. Using as an experimental model organism for the study of cysticercosis, we aimed to identify the conditions necessary to allow parasite development at the short- and long terms. First, cysticerci were incubated for 15 days in different media and parasite densities. The number of buddings and cysticerci diameter were measured to evaluate asexual multiplication and parasite growth, respectively. Vitality was determined by trypan blue staining and morphology analysis. As a result, high cysticerci density and medium containing FBS and the excretion/secretion (E/S) products of feeder cells induced parasite survival, growth and multiplication. Then, the long-term (5 weeks) incubation of the parasites in co-culture with feeder cells was evaluated. Consequently, the mammalian cell lines induced a significant increase in total parasite volume while axenic cultures did not show any statistically significant change over time. In this study, the proper conditions to maintain are described for the first time in a simpler and more controlled setting other than experimental infections. In addition, it was shown that cysticerci growth, survival and asexual multiplication depend on a complex network of secreted factors from both parasite and host.
Topics: Animals; Humans; Mice; Cysticercus; Taenia; Taenia solium; Cysticercosis; Neurocysticercosis; Parasites; Mice, Inbred BALB C; Mammals
PubMed: 36165285
DOI: 10.1017/S0031182022001354 -
Journal of Proteomics Sep 2021Liver fluke, Fascioloides magna, is an important non-native parasite introduced to Europe, posing a threat to survival of local wildlife populations. The aim of this...
Liver fluke, Fascioloides magna, is an important non-native parasite introduced to Europe, posing a threat to survival of local wildlife populations. The aim of this study was to assess the serum and liver protein profile of control and F. magna infected wild boars, by means of shotgun tandem mass tag - based quantitative high resolution proteomics approach. In serum, 4 differentially abundant proteins were found out of total 1073 identified, while in liver from 3520 identified proteins, 116 were differentially abundant between healthy and F. magna infected wild boars. Pathway analysis revealed that most of the proteins differing in abundance are involved in metabolism, biological oxidations, cellular responses to stimuli, fatty acid metabolism, and others. Validation of proteomic results was performed for paraoxonase-1, ceruloplasmin, glutathione S-transferase and liver enzymes by ELISA and automated assays. Complementary analysis of liver and serum in F. magna infection enabled insight into changes of proteome profile of the host at local and sistemic level. Our findings showed that chronic infection with F. magna is associated with immune response in host, oxidative stress and metabolomic changes in liver. SIGNIFICANCE: Liver fluke infections are recognised as worldwide neglected diseases with considerable veterinary and public health importance. Pathological changes, clinical signs and outcome of F. magna infection are strongly related to the type of final hosts and their different tolerance to infection. In order to gain insight into host-parasite interactions in wild boars, dead-end host for F. magna, we assessed proteomics profile of serum and liver of control animals and those infected with F. magna. Proteomics analysis of serum and liver in parallel showed as advantageous and beneficial, demonstrating protein alterations mainly at local level. Bioinformatics analysis enabled elucidation of molecular pathways associated with F. magna infection. Identification and validation of proteins associated with infection may have added value to current tools for efficient liver fluke control.
Topics: Animals; Blood Proteins; Fasciola hepatica; Fasciolidae; Liver; Proteomics; Sus scrofa; Swine; Trematode Infections
PubMed: 34298188
DOI: 10.1016/j.jprot.2021.104332 -
The Journal of Parasitology May 2023The parasites infecting invasive carps in North America (all Cypriniformes: Xenocyprididae: grass carp, Ctenopharyngodon idella [Valenciennes, 1844]; silver carp,...
FIRST RECORD OF A PARASITE, DACTYLOGYRUS CF. SKRJABINI (MONOGENOIDEA: DACTYLOGYRIDAE), INFECTING INVASIVE SILVER CARP, HYPOPHTHALMICHTHYS MOLITRIX (VALENCIENNES, 1844) (CYPRINIFORMES: XENOCYPRIDIDAE) IN NORTH AMERICA.
The parasites infecting invasive carps in North America (all Cypriniformes: Xenocyprididae: grass carp, Ctenopharyngodon idella [Valenciennes, 1844]; silver carp, Hypophthalmichthys molitrix [Valenciennes, 1844]; bighead carp, Hypophthalmichthys nobilis [Richardson, 1845]; and black carp, Mylopharyngodon piceus [Richardson, 1846]) are little studied, and no parasite has been reported from silver carp there. We herein surveyed silver carp from Barkley Reservoir and Cheatham Reservoir (Cumberland River, Tennessee; June and December 2021) and the White River (Arkansas; May 2022) and collected numerous monogenoid specimens infecting the pores on the outer face of the gill raker plate. We heat-killed, formalin-fixed, and routinely stained some specimens for morphology and preserved others in 95% ethanol for DNA extraction and sequencing of the large subunit ribosomal DNA (28S). We identified our specimens as Dactylogyrus cf. skrjabini because they had a dorsal anchor deep root that is much longer than the superficial root, an approximately parallel penis and accessory piece, and a relatively large marginal hook pair V. No type specimen of Dactylogyrus skrjabiniAkhmerov, 1954 (type host and locality is silver carp, Amur River, Russia) is publicly available, but we borrowed several vouchers (NSMT-Pl 6393) that infected the gill rakers of silver carp captured in the Watarase River, Japan. The original description of D. skrjabini was highly stylized and diagrammatical, differing from the specimens we studied from North America and Japan by the dorsal anchor having a superficial root and shaft that comprise a strongly C-shaped hook (the superficial root curves toward the dorsal anchor point) (vs. superficial root straight, at ∼45° angle to deep root and directed away from the dorsal anchor point), a single, much reduced transverse bar that is narrow for its entire breadth (vs. dorsal and ventral transverse bars robust and broad, having an irregular outline), an accessory piece that lacks digitiform projections (vs. accessory piece with 4 digitiform projections), and an accessory piece that lacks a half cardioid-shaped process (vs. accessory piece having a half cardioid-shaped process). Our 28S sequences (generated from 4 specimens of D. cf. skrjabini: 2 from Tennessee [763 base pairs (bp)] and 2 from Arkansas [776 bp]) were identical to 1 ascribed to D. skrjabini from Japan. The present study is the first verifiable and credible report of a parasite from silver carp in North America and the first nucleotide information for a parasite from silver carp in North America.
Topics: Animals; Carps; Helminthiasis, Animal; Platyhelminths; Species Specificity; Tennessee; RNA, Ribosomal, 28S; Phylogeny
PubMed: 37339076
DOI: 10.1645/23-11 -
Molecular and Biochemical Parasitology May 2022Host behavior may be modified by their parasites to increase the likelihood of transmission, but mechanisms underlying these interactions are not well understood. Hosts...
Host behavior may be modified by their parasites to increase the likelihood of transmission, but mechanisms underlying these interactions are not well understood. Hosts and parasites release chemical signaling molecules, like oxylipins, that may affect transmission. Oxylipins are oxygenated metabolites of fatty acids that function as signaling molecules and have essential physiological and functional roles. Yet, the limited taxonomic and contextual scope of these studies constrains our ability to understand their role in parasite-modified behavior. We characterized oxylipins in field-collected File Ramshorn snails, Planorbella pilsbryi. We tested for differences in oxylipin profiles based on infection status (infected with the trematode Echinostoma trivolvis lineage a and uninfected) and parasite activity (high and low). Snail-conditioned water samples were produced by placing five snails into artificial spring water for four hours. Oxylipins were extracted from snail-conditioned water samples and quantified using high performance liquid chromatography-tandem mass spectrometry. Infected snails emitted 69 oxylipins in higher amounts, with 37 only released by this group. Within infected snails, 18 oxylipins were emitted in higher amounts in snails with increased parasite activity. Oxylipins emitted in higher amounts by infected snails with increased parasite activity were predominantly derived from the cytochrome P450 pathway. As infected snails emit different oxylipin profiles than uninfected snails, their production may play a role in altering transmission success. By characterizing the oxylipins produced by snails, and how they are altered by infection, we can test their physiological and ecological roles in freshwater systems.
Topics: Animals; Echinostoma; Host-Parasite Interactions; Oxylipins; Parasites; Trematoda; Water
PubMed: 35227763
DOI: 10.1016/j.molbiopara.2022.111464 -
PloS One 2021Since the first complete set of Platyhelminth nuclear receptors (NRs) from Schistosoma mansoni were identified a decade ago, more flatworm genome data is available to...
Since the first complete set of Platyhelminth nuclear receptors (NRs) from Schistosoma mansoni were identified a decade ago, more flatworm genome data is available to identify their NR complement and to analyze the evolutionary relationship of Platyhelminth NRs. NRs are important transcriptional modulators that regulate development, differentiation and reproduction of animals. In this study, NRs are identified in genome databases of thirty-three species including in all Platyhelminth classes (Rhabditophora, Monogenea, Cestoda and Trematoda). Phylogenetic analysis shows that NRs in Platyhelminths follow two different evolutionary lineages: 1) NRs in a free-living freshwater flatworm (Schmidtea mediterranea) and all parasitic flatworms share the same evolutionary lineage with extensive gene loss. 2) NRs in a free-living intertidal zone flatworm (Macrostomum lignano) follow a different evolutionary lineage with a feature of multiple gene duplication and gene divergence. The DNA binding domain (DBD) is the most conserved region in NRs which contains two C4-type zinc finger motifs. A novel zinc finger motif is identified in parasitic flatworm NRs: the second zinc finger of parasitic Platyhelminth HR96b possesses a CHC2 motif which is not found in NRs of all other animals studied to date. In this study, novel NRs (members of NR subfamily 3 and 6) are identified in flatworms, this result demonstrates that members of all six classical NR subfamilies are present in the Platyhelminth phylum. NR gene duplication, loss and divergence in Platyhelminths are analyzed along with the evolutionary relationship of Platyhelminth NRs.
Topics: Animals; Evolution, Molecular; Gene Duplication; Phylogeny; Platyhelminths; Receptors, Cytoplasmic and Nuclear; Zinc Fingers
PubMed: 34388160
DOI: 10.1371/journal.pone.0250750 -
Methods in Molecular Biology (Clifton,... 2022Spermatogenesis is one of the most dramatic cellular differentiation events observed in animals. In particular, spermiogenesis (the final stage of spermatogenesis)...
Spermatogenesis is one of the most dramatic cellular differentiation events observed in animals. In particular, spermiogenesis (the final stage of spermatogenesis) involves extensive shedding of cytoplasmic organelles, dramatic nuclear rearrangements, and assembly of long flagellar structures. In planarian flatworms, the spherical nucleus present in round spermatids elongates to produce the filamentous nucleus of mature sperm. Newly formed cortical microtubules participate in cytoskeletal rearrangements observed during spermiogenesis and remain present in sperm. In addition, a pair of flagella assemble at one end of each spermatid in a process that likely involves de novo formation of centrioles. This chapter includes a brief introduction to planarian spermatogenesis and current tools for the analysis of molecular players in this process. Step-by-step protocols for isolating and imaging spermatogenic cells are provided with enough detail to be carried out by newcomers to the field who would like to study this unique organism in the laboratory.
Topics: Animals; Cell Nucleus; Male; Microtubules; Morphogenesis; Planarians; Spermatids; Spermatogenesis; Spermatozoa
PubMed: 34542855
DOI: 10.1007/978-1-0716-1661-1_10 -
Parasitology Research Feb 2022Protein phosphorylation plays key roles in a variety of essential cellular processes. Fasciola gigantica is a tropical liver fluke causing hepatobiliary disease...
Protein phosphorylation plays key roles in a variety of essential cellular processes. Fasciola gigantica is a tropical liver fluke causing hepatobiliary disease fascioliasis, leading to human health threats and heavy economic losses. Although the genome and protein kinases of F. gigantica provided new insights to understand the molecular biology and etiology of this parasite, there is scant knowledge of protein phosphorylation events in F. gigantica. In this study, we characterized the global phosphoproteomics of adult F. gigantica by phosphopeptide enrichment-based LC-MS/MS, a high-throughput analysis to maximize the detection of a large repertoire of phosphoproteins and phosphosites. A total of 1030 phosphopeptides with 1244 phosphosites representing 635 F. gigantica phosphoproteins were identified. The phosphoproteins were involved in a wide variety of biological processes including cellular, metabolic, and single-organism processes. Meanwhile, these proteins were found predominantly in cellular components like membranes and organelles with molecular functions of binding (51.3%) and catalytic activity (40.6%). The KEGG annotation inferred that the most enriched pathways of the phosphoproteins included tight junction, spliceosome, and RNA transport (each one contains 15 identified proteins). Combining the reports in other protozoa and helminths, the phosphoproteins identified in this work play roles in metabolic regulation and signal transduction. To our knowledge, this work performed the first global phosphoproteomics analysis of adult F. gigantica, which provides valuable information for development of intervention strategies for fascioliasis.
Topics: Animals; Chromatography, Liquid; Fasciola; Fasciola hepatica; Fascioliasis; Humans; Tandem Mass Spectrometry
PubMed: 34985596
DOI: 10.1007/s00436-021-07422-2 -
Frontiers in Immunology 2022is a common tapeworm in horses causing colic and even mortalities. Current diagnostic tests to detect infections have their limitations and an improved method is...
is a common tapeworm in horses causing colic and even mortalities. Current diagnostic tests to detect infections have their limitations and an improved method is needed. Immunoreactive excretory/secretory proteins (E/S proteome) of this parasite can provide promising candidates for diagnostic tests. We compared E/S proteins produced by small (length < 20 mm, width < 5 mm) and large (length 20 to 40 mm, width 5 to 10 mm) worms by label-free quantitative proteomics using a database composed of related and proteins for protein identifications. Altogether, 509 E/S proteins were identified after incubating the worms for three and eight hours. The greatest E/S proteome changes suggested both worm size- and time-dependent changes in cytoskeleton remodeling, apoptosis, and production of antigens/immunogens. The E/S proteins collected at the three-hour time point represented the natural conditions better than those collected at the eight-hour time point, and thereby contained the most relevant diagnostic targets. Immunoblotting using antibodies from horses tested positive/negative for indicated strongest antigenicity/immunogenicity with 13-, 30- and 100-kDa proteins, involving a thioredoxin, heat-shock chaperone 90 (Hsp90), dynein light chain component (DYNLL), tubulin-specific chaperone A (TBCA) and signaling pathway modulators (14-3-3 and Sj-Ts4). This is among the first studies identifying new diagnostic targets and antigens eliciting a IgG-response in horses.
Topics: Horses; Animals; Proteome; Proteomics; Immunoblotting; Cestoda; Body Fluids; Echinococcus granulosus
PubMed: 36466892
DOI: 10.3389/fimmu.2022.1045468 -
Current Biology : CB Oct 2023Parasites have evolved a variety of astonishing strategies to survive within their hosts, yet the most challenging event in their personal chronicles is the passage from...
Parasites have evolved a variety of astonishing strategies to survive within their hosts, yet the most challenging event in their personal chronicles is the passage from one host to another. It becomes even more complex when a parasite needs to pass through the external environment. Therefore, the free-living stages of parasites present a wide range of adaptations for transmission. Parasitic flatworms from the group Digenea (flukes) have free-living larvae, cercariae, which are remarkably diverse in structure and behavior. One of the cercariae transmission strategies is to attain a prey-like appearance for the host. This can be done through the formation of a swimming aggregate of several cercariae adjoined together by their tails. Through the use of live observations and light, electron, and confocal microscopy, we described such a supposedly prey-mimetic colony comprising cercariae of two distinct morphotypes. They are functionally specialized: larger morphotype (sailors) enable motility, and smaller morphotype (passengers) presumably facilitate infection. The analysis of local read alignments between the two samples reveals that both cercaria types have identical 18S, 28S, and 5.8S rRNA genes. Further phylogenetic analysis of these ribosomal sequences indicates that our specimen belongs to the digenean family Acanthocolpidae, likely genus Pleorchis. This discovery provides a unique example and a novel insight into how morphologically and functionally heterogeneous individuals of the same species cooperate to build colonial organisms for the purpose of infection. This strategy bears resemblance to the cooperating castes of the same species found among insects..
Topics: Humans; Animals; Larva; Parasites; Phylogeny; Swimming; Trematoda; Cercaria
PubMed: 37741283
DOI: 10.1016/j.cub.2023.08.090 -
Parasitology Nov 2022The complete circular mitogenome of (Platyhelminthes: Paragonimidae) from Japan, obtained by PacBio long-read sequencing, was 17 591 bp and contained 12 protein-coding...
The complete circular mitogenome of (Platyhelminthes: Paragonimidae) from Japan, obtained by PacBio long-read sequencing, was 17 591 bp and contained 12 protein-coding genes (PCGs), 2 mitoribosomal RNA and 22 transfer RNA genes. The 8 gene was absent, and there was a 40 bp overlap between 4L and 4. The long non-coding region (4.3 kb) included distinct types of long and short repeat units. The pattern of base usage for PCGs and the mtDNA coding region overall in Asian and American species (, , and ) and the Indian form of was > > > . On the other hand, East-Asian used > > > . Five Asian and American species and had TTT/Phe, TTG/Leu and GTT/Val as the most frequently used codons, whereas the least-used codons were different in each species and between regional forms of . The phylogenetic tree reconstructed from a concatenated alignment of amino acids of 12 PCGs from 36 strains/26 species/5 families of trematodes confirmed that the Paragonimidae is monophyletic, with 100% nodal support. was resolved as a sister to The clade was clearly separate from remaining congeners. The latter clade was comprised of 2 subclades, one of the East-Asian and the other of the Indian Type 1 samples. Additional mitogenomes in the Paragonimidae are needed for genomic characterization and are useful for diagnostics, identification and genetic/ phylogenetic/ epidemiological/ evolutionary studies of the Paragonimidae.
Topics: Animals; Paragonimus; Genome, Mitochondrial; Phylogeny; Trematoda; Lung
PubMed: 36101009
DOI: 10.1017/S0031182022001184