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Scientific Reports Dec 2022The nematode parasite intestine absorbs nutrients, is involved in innate immunity, can metabolize xenobiotics and as we show here, is also a site of action of the...
The nematode parasite intestine absorbs nutrients, is involved in innate immunity, can metabolize xenobiotics and as we show here, is also a site of action of the anthelmintic, diethylcarbamazine. Diethylcarbamazine (DEC) is used to treat lymphatic filariasis and activates TRP-2, GON-2 & CED-11 TRP channels in Brugia malayi muscle cells producing spastic paralysis. DEC also has stimulatory effects on ascarid nematode parasites. Using PCR techniques, we detected, in Ascaris suum intestine, message for: Asu-trp-2, Asu-gon-2, Asu-ced-11, Asu-ocr-1, Asu-osm-9 and Asu-trpa-1. Comparison of amino-acid sequences of the TRP channels of B. malayi, and A. suum revealed noteworthy similarity, suggesting that the intestine of Ascaris will also be sensitive to DEC. We used Fluo-3AM as a Ca indicator and observed characteristic unsteady time-dependent increases in the Ca signal in the intestine in response to DEC. Application of La and the TRP channel inhibitors, 2-APB or SKF 96365, inhibited DEC mediated increases in intracellular Ca. These observations are important because they emphasize that the nematode intestine, in addition to muscle, is a site of action of DEC as well as other anthelmintics. DEC may also enhance the Ca toxicity effects of other anthelmintics acting on the intestine or, increase the effects of other anthelmintics that are metabolized and excreted by the nematode intestine.
Topics: Animals; Ascaris; Ascaris suum; Anthelmintics; Elephantiasis, Filarial; Brugia malayi
PubMed: 36494409
DOI: 10.1038/s41598-022-25648-7 -
Parasitology Nov 2021Due to the presence of artefacts in stool samples, the copromicroscopic diagnosis of Ascaris lumbricoides is not always straightforward, particularly in the case of...
Due to the presence of artefacts in stool samples, the copromicroscopic diagnosis of Ascaris lumbricoides is not always straightforward, particularly in the case of fertilized decorticated eggs. A total of 286 stool samples from 115 schoolchildren in India and 171 adult immigrants in Italy were screened for the presence of A. lumbricoides eggs by both Kato-Katz thick smear and Mini-FLOTAC. If the outer layer of A. lumbricoides eggs was absent, two aliquots of each stool sample were preserved: one for coproculture to identify larvae after development and one to compose a pool of stool for molecular analysis. A total of 64 stool samples (22.4%) were positive for A. lumbricoides using the Kato-Katz thick smear; 36 (56.3%) of these showed mammillated A. lumbricoides eggs, 25 (39.1%) showed elements resembling fertilized decorticated eggs, while three samples (4.7%) showed both mammillated and decorticated eggs. By Mini-FLOTAC, 39 stool samples (13.6%) were positive, while decorticated A. lumbricoides-like eggs were identified as artefacts. These results were confirmed by negative coprocultures and quantitative polymerase chain reaction. Mini-FLOTAC can be used for a reliable diagnosis of A. lumbricoides, thanks to the flotation and translation features which allow a clearer view, resulting in the correct identification of A. lumbricoides eggs.
Topics: Animals; Artifacts; Ascaris lumbricoides; Feces; Helminthiasis; Sensitivity and Specificity
PubMed: 34250886
DOI: 10.1017/S0031182021001256 -
Nature Communications Feb 2015Toxocara canis is a zoonotic parasite of major socioeconomic importance worldwide. In humans, this nematode causes disease (toxocariasis) mainly in the under-privileged...
Toxocara canis is a zoonotic parasite of major socioeconomic importance worldwide. In humans, this nematode causes disease (toxocariasis) mainly in the under-privileged communities in developed and developing countries. Although relatively well studied from clinical and epidemiological perspectives, to date, there has been no global investigation of the molecular biology of this parasite. Here we use next-generation sequencing to produce a draft genome and transcriptome of T. canis to support future biological and biotechnological investigations. This genome is 317 Mb in size, has a repeat content of 13.5% and encodes at least 18,596 protein-coding genes. We study transcription in a larval, as well as adult female and male stages, characterize the parasite's gene-silencing machinery, explore molecules involved in development or host-parasite interactions and predict intervention targets. The draft genome of T. canis should provide a useful resource for future molecular studies of this and other, related parasites.
Topics: Animals; Dogs; Female; Gene Silencing; Genome Size; Genome, Helminth; High-Throughput Nucleotide Sequencing; Host-Parasite Interactions; Humans; Larva; Male; Microsatellite Repeats; Molecular Sequence Annotation; Open Reading Frames; Phylogeny; Toxocara canis; Toxocariasis; Transcriptome
PubMed: 25649139
DOI: 10.1038/ncomms7145 -
Scientific Reports Aug 2020P-glycoproteins (Pgp) have been proposed as contributors to the widespread macrocyclic lactone (ML) resistance in several nematode species including a major pathogen of...
P-glycoproteins (Pgp) have been proposed as contributors to the widespread macrocyclic lactone (ML) resistance in several nematode species including a major pathogen of foals, Parascaris univalens. Using new and available RNA-seq data, ten different genomic loci encoding Pgps were identified and characterized by transcriptome-guided RT-PCRs and Sanger sequencing. Phylogenetic analysis revealed an ascarid-specific Pgp lineage, Pgp-18, as well as two paralogues of Pgp-11 and Pgp-16. Comparative gene expression analyses in P. univalens and Caenorhabditis elegans show that the intestine is the major site of expression but individual gene expression patterns were not conserved between the two nematodes. In P. univalens, PunPgp-9, PunPgp-11.1 and PunPgp-16.2 consistently exhibited the highest expression level in two independent transcriptome data sets. Using RNA-Seq, no significant upregulation of any Pgp was detected following in vitro incubation of adult P. univalens with ivermectin suggesting that drug-induced upregulation is not the mechanism of Pgp-mediated ML resistance. Expression and functional analyses of PunPgp-2 and PunPgp-9 in Saccharomyces cerevisiae provide evidence for an interaction with ketoconazole and ivermectin, but not thiabendazole. Overall, this study established reliable reference gene models with significantly improved annotation for the P. univalens Pgp repertoire and provides a foundation for a better understanding of Pgp-mediated anthelmintic resistance.
Topics: ATP Binding Cassette Transporter, Subfamily B, Member 1; Animals; Antiparasitic Agents; Ascaridida Infections; Ascaridoidea; Drug Resistance; Helminth Proteins; Horses; Ivermectin; Phylogeny; Reverse Transcriptase Polymerase Chain Reaction; Sequence Analysis, DNA; Sequence Analysis, RNA; Transcriptome
PubMed: 32788636
DOI: 10.1038/s41598-020-70529-6 -
BMC Veterinary Research Mar 2020Wild Amur tigers are a sparsely populated species, and the conservation of this species is of great concern worldwide, but as an important health risk factor, parasite...
BACKGROUND
Wild Amur tigers are a sparsely populated species, and the conservation of this species is of great concern worldwide, but as an important health risk factor, parasite infection in them is not fully understanding.
RESULTS
In this study, sixty-two faecal samples were collected to investigate the frequency and infection intensity of Toxocara cati and Toxascaris leonina in wild Amur tigers. The T. cati and T. leonina eggs were preliminary identified by microscopy, and confirmed by molecular techniques. Infection intensity was determined by the modified McMaster technique. Phylogenetic trees demonstrated that T. cati of wild Amur tiger had a closer relationship with which of other wild felines than that of domestic cats. T. leonina of Amur tiger and other felines clustered into one clade, showing a closer relationship than canines. The average frequency of T. cati was 77.42% (48/62), and the frequency in 2016 (100%) were higher than those in 2013 (P = 0.051, < 0.1; 66.6%) and 2014 (P = 0.079, < 0.1; 72.2%). The infection intensity of T. cati ranged from 316.6 n/g to 1084.1 n/g. For T. leonina, only three samples presented eggs when the saturated sodium chloride floating method was performed, indicating that the frequency is 4.83% (3/62). Unfortunately, the egg number in faecal smears is lower than the detective limitation, so the infection intensity of T. leonina is missed.
CONCLUSIONS
This study demonstrated that ascarids are broadly prevalent, and T. cati is a dominant parasite species in the wild Amur tiger population.
Topics: Animals; China; Parasite Egg Count; Phylogeny; Tigers; Toxascariasis; Toxascaris; Toxocara; Toxocariasis
PubMed: 32156273
DOI: 10.1186/s12917-020-02296-5 -
Systematic Parasitology Oct 2017Ingwenascaris n. g. (Nematoda: Heterocheilidae) is established to accommodate Ingwenascaris sprenti n. g., n. sp., described from the stomach of Crocodylus niloticus...
Ingwenascaris n. g. (Nematoda: Ascaridida: Heterocheilidae) established for I. sprenti n. sp. and I. assymmetrica (Ortlepp, 1932) n. comb., parasites of African crocodiles, and an identification key to the genera of the Heterocheilidae.
Ingwenascaris n. g. (Nematoda: Heterocheilidae) is established to accommodate Ingwenascaris sprenti n. g., n. sp., described from the stomach of Crocodylus niloticus Laurenti (Reptilia: Crocodylidae) in South Africa, based on light and scanning electron microscopy studies of its morphology. The new genus can be distinguished from other heterocheilid genera through a combination of its characters, including the pronounced asymmetry of each subventral lip due to an alate ventral margin and a non-alate margin facing the dorsal lip, the presence of continuous ridges of triangular denticles along the free labial margins, the lack of interlocking processes or a rostral plate, interlabia being indistinct or represented by small lateral interlabia between the dorsal and ventral lips only, the absence of prominent interlabial longitudinal cuticular ridges, the presence of lateral alae that are fused with the subventral lips, the presence of lateral caudal alae in both sexes, spicules of males that are composed of handle and alate blade, the presence of a gubernaculum, the number and arrangement of male caudal papillae and the position of the vulva near the anterior and middle third of the body in females. Ingwenascaris sprenti n. g., n. sp. represents the sixth heterocheilid genus parasitising African crocodilians. Trispiculascaris assymmetrica (Ortlepp, 1932) (syn. Porrocaecum assymmetricum Ortlepp, 1932) from a Central African crocodile is transferred to the new genus as I. assymmetrica (Ortlepp, 1932) n. comb. The genus Trispiculascaris Skrjabin, 1916 is considered a genus incertae sedis. An identification key to the genera of the family Heterocheilidae is presented.
Topics: Alligators and Crocodiles; Animals; Ascaridida; South Africa; Species Specificity; Stomach
PubMed: 28864918
DOI: 10.1007/s11230-017-9748-y -
Parasitology Research Jun 2023Toxocara tanuki is a common large roundworm in raccoon dogs. Experimental infection studies of T. tanuki in mice were conducted to clarify the distribution and...
Toxocara tanuki is a common large roundworm in raccoon dogs. Experimental infection studies of T. tanuki in mice were conducted to clarify the distribution and infectivity of larvae in tissue. Groups of BALB/c and C57BL/6 mice (n = 5 mice/group) were each inoculated with 1000 embryonated T. tanuki eggs and necropsied at 7, 31, 91, and 182 days post inoculation (dpi). The number of larvae in the central nervous system, heart, lungs, kidneys, spleen, gastrointestinal tract, liver, and carcass was examined. Larvae obtained from the aforementioned mice on different days of the necropsy were orally inoculated into four groups of ICR mice (n = 6 mice/group) that were then necropsied at 21 dpi. Larvae were recovered from all mice. In the BALB/c and C57BL/6 mice, most of the larvae (> 88.7%) were recovered from the liver and the remainder from other tissues. The total number of larvae recovered from C57BL/6 mice was significantly higher than that from BALB/c mice, but no difference in the relative larval distribution within the viscera between the two mouse strains was observed. The mean recovery percentage of larvae from ICR mice infected with 182-day-old tissue larvae was 3.3%. Our findings showed that T. tanuki larvae migrated predominantly to the liver of mice and that the larvae maintained their infectivity for at least half a year.
Topics: Animals; Mice; Toxocara; Toxocariasis; Larva; Mice, Inbred ICR; Mice, Inbred C57BL; Lung; Liver; Toxocara canis
PubMed: 37046027
DOI: 10.1007/s00436-023-07832-4 -
Parasites & Vectors Jun 2023The roundworms, Parascaris spp., are important nematode parasites of foals and were historically model organisms in the field of cell biology, leading to many important...
BACKGROUND
The roundworms, Parascaris spp., are important nematode parasites of foals and were historically model organisms in the field of cell biology, leading to many important discoveries. According to karyotype, ascarids in Equus are commonly divided into Parascaris univalens (2n = 2) and Parascaris equorum (2n = 4).
METHODS
Here, we performed morphological identification, karyotyping and sequencing of roundworms from three different hosts (horses, zebras and donkeys). Phylogenetic analysis was performed to study the divergence of these ascarids based on cytochrome c oxidase subunit I (COI) and internal transcribed spacer (ITS) sequences.
RESULTS
Karyotyping, performed on eggs recovered from worms of three different Equus hosts in China, showed two different karyotypes (2n = 2 in P. univalens collected from horses and zebras; 2n = 6 in Parascaris sp. collected from donkeys). There are some differences in the terminal part of the spicula between P. univalens (concave) and Parascaris sp. (rounded). Additionally, it was found that the egg's chitinous layer was significantly thicker in Parascaris sp. (> 5 μm) than P. univalens (< 5 μm) (F = 1967, P < 0.01). Phylogenetic trees showed that the sequences of Parascaris from Equus hosts were divided into two distinct lineages based on sequences of the COI and ITS.
CONCLUSIONS
Comparing the differences in roundworms collected from three different Equus hosts, this study describes a Parascaris species (Parascaris sp.) with six chromosomes in donkeys. It is worth noting that the thickness of the chitinous layer in the Parascaris egg may serve as a diagnostic indicator to distinguish the two roundworms (P. univalens and Parascaris sp.). The Parascaris sp. with six chromosomes in donkeys in the present study may be a species of P. trivalens described in 1934, but the possibility that it is a new Parascaris species cannot be ruled out. Both karyotyping and molecular analysis are necessary to solve the taxonomic problems in Parascaris species.
Topics: Horses; Animals; Ascaridoidea; Phylogeny; Ascaridida Infections; Horse Diseases; Equidae; China
PubMed: 37322493
DOI: 10.1186/s13071-023-05768-3 -
PLoS Neglected Tropical Diseases Apr 2024The World Health Organization emphasizes the importance of integrated monitoring and evaluation in neglected tropical disease (NTD) control programs. Serological assays... (Review)
Review
BACKGROUND
The World Health Organization emphasizes the importance of integrated monitoring and evaluation in neglected tropical disease (NTD) control programs. Serological assays offer a potential solution for integrated diagnosis of NTDs, particularly for those requiring mass drug administration (MDA) as primary control and elimination strategy. This scoping review aims (i) to provide an overview of assays using serum or plasma to detect infections with soil-transmitted helminths (STHs) in both humans and animals, (ii) to examine the methodologies used in this research field and (iii) to discuss advancements in serological diagnosis of STHs to guide prevention and control programs in veterinary and human medicine.
METHODOLOGY
We conducted a systematic search in the Ovid MEDLINE, Embase and Cochrane Library databases, supplemented by a Google search using predefined keywords to identify commercially available serological assays. Additionally, we performed a patent search through Espacenet.
PRINCIPAL FINDINGS
We identified 85 relevant literature records spanning over 50 years, with a notable increased interest in serological assay development in recent years. Most of the research efforts concentrated on diagnosing Ascaris infections in both humans and pigs, primarily using ELISA and western blot technologies. Almost all records targeted antibodies as analytes, employing proteins and peptides as analyte detection agents. Approximately 60% of sample sets described pertained to human samples. No commercially available tests for Trichuris or hookworms were identified, while for Ascaris, there are at least seven different ELISAs on the market.
CONCLUSIONS
While a substantial number of assays are employed in epidemiological research, the current state of serological diagnosis for guiding STH prevention and control programs is limited. Only two assays designed for pigs are used to inform efficient deworming practices in pig populations. Regarding human diagnosis, none of the existing assays has undergone extensive large-scale validation or integration into routine diagnostics for MDA programs.
Topics: Humans; Animals; Swine; Ancylostomatoidea; Trichuris; Ascaris; Soil; Ascaris lumbricoides; Helminths; Feces; Helminthiasis; Prevalence
PubMed: 38574166
DOI: 10.1371/journal.pntd.0012049 -
Environmental Science and Pollution... Mar 2021The present study is aimed at assessing the effectiveness of solar drying process in terms of helminth egg reduction in sewage sludge (SS) generated from an activated...
The present study is aimed at assessing the effectiveness of solar drying process in terms of helminth egg reduction in sewage sludge (SS) generated from an activated sludge wastewater treatment plant (WWTP) in Marrakesh city (Morocco). It is also engaged to highlight a synergic effect of liming (1% CaO) and solar drying on helminth egg reduction. The solar drying process was conducted for 45 days, in summer under a semi-arid climate in a pilot scale polycarbonate-based tunnel (2 m). Before undergoing solar drying process, data showed an important load of helminth eggs including Ascaris sp., Schistosoma spp., Capillaria spp., Trichuris spp., Ankylostome spp., Toxocara spp., and Taenia spp. in limed sludge (LS) and non-limed sludge (NLS) (15.2 and 17.9 eggs/g, respectively). Ascaris eggs were the most abundant (11.2 and 13.5 eggs/g in LS and NLS, respectively). By the end of the solar drying process, a considerable removal of the total helminth eggs was recorded in LS and NLS (92.8% and 91.6%, respectively). A complete removal of Schistosoma spp., Capillaria spp., Trichuris spp., Toxocara spp. and Taenia spp. was noted in LS and NLS. In the case of Ankylostome spp., data showed a total removal in LS and 81% in NLS; however, the final load is in agreement with the standards (0.4 egg/g). As for Ascaris spp., neither liming nor solar drying process allowed a complete removal (91% and 90% in NLS and LS, respectively) and the final load (1.1 egg/g) does not fulfill the WHO requirements for an agricultural use. Principal component analysis (PCA) demonstrated a negative correlation between dry matter (DM) content (hence temperature) and helminth egg concentration. No significant synergic effect of liming and solar drying process was showed by statistical analysis. This is substantiating that temperature is the key parameter involved in helminth egg removal while undergoing solar drying of SS.
Topics: Animals; Ascaris; Helminths; Morocco; Sewage; Toxocara
PubMed: 33201505
DOI: 10.1007/s11356-020-11619-w