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Chemical mutagenesis of the parasitic nematode Strongyloides ratti to isolate ivermectin resistant mutants.

We describe a strategy for the mutagenesis of the free-living adult generation of Strongyloides ratti and selection of worms carrying new mutations in the subsequent F2 generation of infective larvae. We demonstrate that this strategy is successful via the selection of infective larvae that are resistant to the anthelmintic ivermectin at a concentration of 10 ng/ml. The majority of these larvae were unable to give rise to patent infections when used to infect parasite naive rats, implying that the majority of the ivermectin resistance mutations confer pleiotropic defects on parasitic, but not on free-living, development.

Animals↗

The mitochondrial genome of Strongyloides stercoralis (Nematoda) - idiosyncratic gene order and evolutionary implications.

The complete mitochondrial genome sequence of the parasitic nematode Strongyloides stercoralis was determined, and its organisation and structure compared with other nematodes for which complete mitochondrial sequence data were available. The mitochondrial genome of S. stercoralis is 13,758 bp in size and contains 36 genes (all transcribed in the clockwise direction) but lacks the atp8 gene. This genome has a high T content (55.9%) and a low C content (8.3%). Corresponding to this T content, there are 16 (poly-T) tracts of >/=12 Ts distributed across the genome. In protein-coding genes, the T bias is greatest (76.4%) at the third codon position compared with the first and second codon positions. Also, the C content is higher at the first (9.3%) and second (13.4%) codon positions than at the third (2%) position. These nucleotide biases have a significant effect on predicted codon usage patterns and, hence, on amino acid compositions of the mitochondrial proteins. Interestingly, six of the 12 protein-coding genes are predicted to employ a unique initiation codon (TTT), which has not yet been reported for any other animal mitochondrial genome. The secondary structures predicted for the 22 transfer RNA (trn) genes and the two ribosomal RNA (rrn) genes are similar to those of other nematodes. In contrast, the gene arrangement in the mitochondrial genome of S. stercoralis is different from all other nematodes studied to date, revealing only a limited number of shared gene boundaries (atp6-nad2 and cox2-rrnL). Evolutionary analyses of mitochondrial nucleotide and amino acid sequence data sets for S. stercoralis and seven other nematodes demonstrate that the mitochondrial genome provides a rich source of phylogenetically informative characters. In conclusion, the S. stercoralis mitochondrial genome, with its unique gene order and characteristics, should provide a resource for comparative mitochondrial genomics and systematics studies of parasitic nematodes.

Amino Acid Sequence↗

Structure and developmental expression of Strongyloides stercoralis fktf-1, a proposed ortholog of daf-16 in Caenorhabditis elegans.

A forkhead transcription factor gene, fktf-1, which we propose to be orthologous to the Caenorhabditis elegans dauer-regulatory gene daf-16 has been discovered in the parasitic nematode Strongyloides stercoralis. Genomic and cDNA sequences from both species predict alternately spliced a and b message isoforms. In contrast to C. elegans, where two a isoforms, daf-16a1 and daf-16a2, are found, a single fktf-1a isoform is found in S. stercoralis. Five of the 10 introns found in the C. elegans gene are found in the proposed S. stercoralis ortholog. Functional motifs common to DAF-16 and several mammalian forkhead transcription factors are conserved in FKTF-1. These include the forkhead DNA binding domain, four Akt/protein kinase B phosphorylation sites and a C-terminal domain that may associate with factors such as the steroid receptor coactivator and other factors necessary for transcriptional regulation. An N-terminal serine-rich domain found in DAF-16A is greatly expanded in FKTF-1A. This domain is missing in DAF-16B, FKTF-1B and all mammalian orthologs. FKTF-1 shows the closest phylogenetic relationship to DAF-16 among all known mammalian and nematode forkhead transcription factors. Like its proposed Caenorhabditis ortholog, the fktf-1 message is expressed at all stages of the life cycle examined thus far. Discovery of fktf-1 indicates the presence of an insulin-like signalling pathway in S. stercoralis similar to that known to regulate dauer development in C. elegans. This pathway is a likely candidate to control infective larval arrest and reactivation as well as regulation of the switch between parasitic and free-living development in the parasite.

Animals↗

Strongyloides stercoralis: oral transfer of parasitic adult worms produces infection in mice and infection with subsequent autoinfection in gerbils.

Oral transfer of parasitic adult Strongyloides stercoralis produced patent infections in gerbils, C57BL/6J and SCID mice. In gerbils receiving adult worms, 7.3% of the transferred worms established and autoinfective L3 were found beginning on day 5 post-transfer, with peak numbers seen on days 6 and 7 post-transfer and few seen by 9 days post-transfer. These results suggest that development of autoinfective L3 in the gerbil is limited by the immune response of the host. When given orally to mice, between 7.2% (C57BL/6J) and 19.5% (SCID) of the adult worms established. These levels are higher than those previously obtained by the subcutaneous infection of SCID mice with infective larvae. No autoinfective larvae were found in infected mice and the ratio of L1/adult worms was small compared with that seen in gerbils. Thus, mice infected orally can be used as a model to study the interaction between the adult worm and the host, and since autoinfection has not been seen in the murine model, as developed to date, orally infected mice may be useful as a model to study mechanisms preventing autoinfection.

Animals↗

Gelatin particle indirect agglutination and enzyme-linked immunosorbent assay for diagnosis of strongyloidiasis using Strongyloides venezuelensis antigen.

Routine microscopical examination of stool specimens for diagnosis of strongyloidiasis is insensitive and serological methods using Strongyloides stercoralis antigen are at present not available for field studies. We evaluated 2 techniques, enzyme-linked immunosorbent assay (ELISA) and gelatin particle indirect agglutination (GPIA), using an antigen obtained from the rodent parasite, S. venezuelensis. Fifty-four Peruvian patients with different clinical forms of strongyloidiasis were studied: 12 asymptomatic, 31 symptomatic, and 11 hyperinfection cases. Our results demonstrate that both ELISA and GPIA using S. venezuelensis antigen are useful for diagnosis of strongyloidiasis, with sensitivities of 74.1% and 98.2%, respectively and a specificity of 100% for both techniques. We found that GPIA is a highly sensitive test for patients with suspected chronic infection and/or hyperinfection. In the hyperinfection cases, significantly lower concentrations of specific immunoglobulin antibodies and eosinophils (P < 0.001) were found compared with the asymptomatic and symptomatic cases.

Adolescent↗

Epidemiology of Strongyloides stercoralis in north-east Thailand: application of the agar plate culture technique compared with the enzyme-linked immunosorbent assay.

Cross-sectional surveys of parasitic infection were performed using the agar plate culture technique (APCT) and modified formalin-ethyl acetate concentration technique (MFECT) to assess the true prevalence of Strongyloides stercoralis relative to other parasites in north-east Thailand. The enzyme-linked immunosorbent assay (ELISA) for diagnosis of S. stercoralis infection was used to estimate the seroprevalence for comparison with coproprevalence. Faecal and serum samples were collected from study participants during October-November 2000. Within the sample population of 332 rural northeast Thais from 3 communities, S. stercoralis was the most common parasitic infection (average 28.9%, range 27.7-30.3%) as determined by APCT; by MFECT the average was 5.4% (range 1.8-8.6%). Other intestinal parasites by order of prevalence were Opisthorchis viverrini (average 14.2%, range 8.6-19.4%), hookworm (average 12.3%, range 4-20.2%), Echinostoma sp. (7.5%), Giardia intestinalis (0.9%), Trichuris trichiura (0.6%), and Taenia sp., Hymenolepis nana and Entamoeba coli (all 0.3%). In an analysis of a subset of the sample population for which serum samples were available (n = 120), coproprevalence by APCT was 33.3% (range 27-53.8%) and seroprevalence was 47.5% (range 29.7-57.9%) by modified unit-based ELISA and 34.2% (range 21.6-42.1%) by conventional optical density (OD)-based ELISA. Taking APCT as the reference method for diagnosis of strongyloidiasis, the sensitivity and specificity of the OD-based ELISA were 65% and 81.3%, respectively, and of the unit-based ELISA were 77.5% and 71.3%, respectively. Our results indicate that S. stercoralis is the predominant parasite in rural north-east Thailand, and that APCT and ELISA should be used as complementary diagnostic methods for community-based parasite surveys, at least among those in high-risk groups.

Adolescent↗

Increased sensitivity of routine laboratory detection of Strongyloides stercoralis and hookworm by agar-plate culture.

The efficacy of agar-plate culture has been evaluated for the detection of Strongyloides stercoralis and hookworm, compared with direct smear, the formalin-ether sedimentation technique and the filter-paper method. Of 1085 stool samples from the routine laboratory service at King Chulalongkorn Memorial Hospital in Bangkok, 241 samples harboured S. stercoralis, 153 hookworm and 2 Rhabditis hominis. The recovery rate of S. stercoralis by agar-plate culture is significantly superior to the other methods (P < 0.005). The ratios of positive results from the methods used to the total number of S. stercoralis-positive cases were as follows: 1:1.03 by agar-plate culture, 1:1.85 by the filter-paper method, 1:1.98 by the sedimentation technique and 1:10.48 by direct stool smear. A similar trend of the efficacy ratio of each method was obtained for hookworm detection. The characteristic furrows left by hookworm larvae, and larvae and adults of S. stercoralis could be used for preliminary species identification. Daily search for furrows on agar plates for up to 6 consecutive days resulted in an increased sensitivity for diagnosis of both S. stercoralis and hookworm infections.

Adolescent↗

Sub-acute intestinal obstruction by Strongyloides stercolaris.

Strongyloides stercolaris infestation is rather rare in Iraq. Individuals with infection confined to the intestinal tract are often asymptomatic. Symptoms include abdominal pain, diarrhea, weight loss and other non-specific complaints. The diagnosis depends upon repeated examination of stool and duodenal aspirate. Two cases presenting as sub-acute intestinal obstruction and mimicking primary intestinal lymphoma (PIL) on presentation are presented. Differentiation between the two conditions regarding presenting features, barium studies and pathology are discussed.

Adult↗

Exploiting the life cycle of Strongyloides ratii.

The nematode Strongyloides ratti has a remarkable life cycle, which has both a parasitic and a free-living phase. The free-living phase includes a choice between two developmental routes. Here, Mark Viney discusses recent advances in understanding the biology of this developmental switch and shows how the life cycle of this nematode can be used to explore the lifestyle transitions common to all parasitic nematodes, as well as to address other basic biological questions.

Animals↗

Strongyloides stercoralis infection presenting as generalized prurigo nodularis and lichen simplex chronicus.

Strongyloides Stercoralis is a parasitic nematode that develops an autoinfective life cycle within the gastrointestinal tract of its human host. The infection produces peripheral eosinophilia and cutaneous eruptions, as well as gastrointestinal or respiratory symptoms. Detection of S stercoralis is difficult through stool examination, but may be demonstrated by ELISA for IgG antibody against the parasite. We describe a patient with chronic S stercoralis infection initially presenting with generalized prurigo nodularis and lichen simplex chronicus.

Adult↗

[Strongyloides stercoralis infection in patients with bronchial obstructive pathology].

Strongyloides stercoralis infection was hardly seen in Spain until a few years ago but has recently been shown to be fairly common in some geographic areas. In the respiratory tract this germ can cause acute bronchospasms that make diagnosis difficult, particularly in patients with underlying bronchial disease. To determine if curing S. stercoralis infection is accompanied by clinical and functional improvement in patients with bronchial obstruction, we studied the evolution of 22 infected patients: 11 with no bronchopulmonary disease and 11 with chronic airway obstruction or asthma. The following variables were assessed in both groups at the moment of diagnosis of infection and four months after cure: levels of eosinophils and total serum IgE, respiratory symptoms, steroid doses and spirometric parameters. After four months we observed a significant decrease in eosinophil (16 versus 5%) and IgE (1,600 versus 770 IU/ml) levels in both groups. The number of bronchospasms and daily steroid doses required decreased in the group with bronchial disease. No significant differences were seen in spirometric parameters, however. The improvement in respiratory symptoms, blood parameters and need for medication leads us to believe that airway inflammation decreases after the infection has been eradicated, in spite of the lack of improvement in bronchial obstruction.

Aged↗

[Strongyloides stercoralis infection in a cortico-dependent patient with chronic airflow obstruction].

We report a case of Strongyloides stercoralis infection in a male agricultural worker with corticoid dependent chronic obstructive air flow disease and bronchial hyperreactivity. The clinical picture initially involved digestive symptoms and bronchospasm resistant to steroid treatment. Diagnosis was established by demonstrating the larvae of S. stercoralis in stool. We emphasize that this infection should be suspected in patients living in endemic areas such as Spain, and describe S. stercoralis-caused respiratory manifestations that can be masked by bronchial disease.

Aged↗

Strongyloides stercoralis infection: how to diagnose best?

Four patients are described with a Strongyloides stercoralis infection. Several techniques to diagnose this infection are discussed. The so-called Baermann method is emphasised. Especially in chronic infections the combination of serology and the Baermann method seems the best diagnostic approach. Treatment with albendazole or ivermectin are suggested treatments.

Adult↗

Characterization of the protective response against a homologous challenge infection with Strongyloides venezuelensis in rats.

The protective response in rats against a homologous challenge infection with Strongyloides venezuelensis was characterized. In an initial infection with 1000 filariform larvae and migrating larvae (L(3)) of S. venezuelensis, the population of L(3) in the lungs on day 3 postinfection (PI), and that of adult worms in the small intestine on day 7 PI, were 180.8+/-14.5 and 336.8+/-70.7, respectively. The latter were gradually expelled towards day 42 PI. After the initial infection, the rats developed strong immunity against a homologous challenge infection as manifested by a marked reduction in worm populations, stunted body length and width, damage to reproductive organs, impaired egg production and rapid expulsion of the worms by day 14 after challenge. Expulsion of the worms was preceded by a significantly elevated (P<0.05) peripheral blood eosinophil (PBE) count, both in the initial (200.0+/-26.5 x 10(3)ml) and the challenge infection (400.9+/-165.4 x 10(3)ml). These findings suggest that rats acquire strong homologous immunity following initial exposure to S. venezuelensis. It is suggested that PBEs are involved in worm expulsion. A major target of these effector mechanisms is the reproductive system of S. venezuelensis.

Animals↗

The control of the free-living stages of Strongyloides papillosus by the nematophagous fungus, Arthrobotrys oligospora.

Two laboratory trials were conducted to determine the effect of the addition of spores (conidia) of the nematophagous fungus, Arthrobotrys oligospora, on the development of the ruminant parasite, Strongyloides papillosus, in cultures of bovine faeces. Both studies showed that at a concentration of 2000 conidia/g faeces virtually eliminated infective larvae (> 99% reduction), following 14 days incubation under ideal conditions (25 degrees C and saturated humidity) for free-living development of this parasite species. In one trial, a high level of control was also observed at a 10-fold decrease in conidia concentration (200 spores/g faeces). This work has demonstrated, in principle, that A. oligospora could provide a practical biological control agent against S. papillosus infecting intensively raised young ruminants in the humid tropics/subtropics.

Animals↗

Hypolipemia associated with the wasting condition of rabbits infected with Strongyloides papillosus.

Rabbits develop a wasting condition in the intestinal stage of Strongyloides papillosus infection. Serum inflammatory cytokine and lipid profiles were investigated in five rabbits infected with S. papillosus and five uninfected pair-fed controls to ascertain whether the disease is inflammatory cytokine-mediated cachexia. Tumor necrosis factor alpha (TNF alpha) was detected in one infected animal at Day 7 after infection. Interleukin (IL)-1 was detected in three infected, and one control, animals at Day 28. IL-6 remained unchanged in both the groups. Infected animals developed hypolipemia, including hypotriglyceridemia in the intestinal stage of infection. Control animals lost body weight in the same manner as the infected animals, but had elevated cholesterols and phospholipids with normal triglyceride concentrations. The results suggested that the wasting condition has no association with cachexia induced by TNF alpha. IL-1 or IL-6, and that hepatic function for lipid synthesis is affected during the intestinal stage of S. papillosus infection.

Animals↗

Mesenteric arteriographic findings in a patient with strongyloides stercoralis hyperinfection.

The authors present a case of a Latin American patient with systemic lupus erythematosus who was referred for a mesenteric arteriogram because of acute lower gastrointestinal bleeding. Multiple segments of dilation alternating with stenosis or spasm were noted in the superior mesenteric artery/inferior mesenteric artery distributions. At the time, these irregularities were thought to be representative of lupus vasculitis. Despite appropriate treatment for vasculitis, the patient continued to have bleeding episodes and ultimately died of multiple organ failure. Autopsy demonstrated no evidence of vasculitis, but did demonstrate the unexpected finding of Strongyloides stercoralis hyperinfection with vessel invasion.

Adult↗

Adult worms of the rodent intestinal nematode, Strongyloides venezuelensis, successfully invade chick intestinal mucosa.

In order to study the mucosal invasion of a rodent intestinal nematode in bird intestine, chicks were infected with the intestinal nematode of rodents, Strongyloides venezuelensis, by subcutaneous larva inoculation and adult worm implantation. No evidence was obtained for larvae reaching the lungs or the intestine after infective larva inoculation. Adult worms implanted in the small intestine invaded the mucosa and remained there at least for 24 h, whereas those implanted in the caecum were trapped by mucus, and did not invade the mucosa. Mucosal invasion of adult worms in the small intestine was confirmed by histological examination. The number of adult worms in the intestinal mucosal tissue dropped rapidly within the first 24 h, which was associated with infiltrating granulocytes around the worms. The present study suggests that S. venezuelensis adult worms are able to invade the intestinal tissue of chicks, which do not belong to the vertebrate class of its normal definitive host, but that they are eliminated rapidly by mucosal defense system of the bird.

Animals↗