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An alternative protocol for the serial passage and maintenance of a homogonic strain of Strongyloides ratti in female Sprague-Dawley rats.

A method which does not involve the tedious use of watch glass coprocultures for obtaining filariform infective (L3) larvae of Strongyloides ratti from faecal pellets of infected Sprague-Dawley rats is described. The alternative method utilises Baermannization (18 h) of faecal pellets to yield rhabditiform (L1) larvae of S. ratti and their subsequent culture for 72 h at 19 degrees C in tissue-culture-flasks containing only dechlorinated tap water to yield infective filariform (L3) larvae. The yields and infectivity of the L3 larvae obtained from the two methods were essentially similar.

Animals↗

Analysis of the 18S ribosomal RNA gene of Strongyloides stercoralis.

The entire 1766 bases of the 18S rRNA gene of Strongyloides stercoralis have been sequenced. The gene has a 38% G+C content. Although it is similar in length to the 18S rRNA gene of Caenorhabditis elegans, the only other completely sequenced nematode 18S rRNA gene, it is only 69% identical. Closely related helminths will need to be sequenced in order to delineate sequences specific for the diagnosis of strongyloidiasis.

Animals↗

Regulatory effect of anti-interleukin-5 monoclonal antibody on intestinal worm burden in a primary infection with strongyloides venezuelensis in mice.

The intestinal worm burden in Strongyloides venezuelensis-infected mice was influenced by treatment with anti-interleukin-5 (IL-5) monoclonal antibody (NC17) when NC17 was given to mice 3-7 days before infection. The present study has examined the involvement of IL-5 in susceptibility at different in the development of the parasite in the host. The results show that the number of tissue-migrating larvae recovered from the lungs in a primary infection was not affected by anti-IL-5 monoclonal antibody treatment, whereas intestinal worm counts increased in mice treated with 0.25-1 mg of NC17. In mice treated with 0.1 mg of NC17, adult worm recovery was not significantly different from non-treated controls. Peripheral and tissue eosinophilia were not observed in the early phase of infection (days 4-8). Six days after transfer of lung-stage larvae to NC17-treated mice, adult worm recovery was higher than that of control mice. These results suggest that non-eosinophil response(s), which were dependent on IL-5, were involved in the initial establishment of the intestinal stage of S. venezuelensis in mice. We discuss the mechanisms that control the susceptibility to the parasite from the viewpoint of host defence.

Animals↗

Steady-state content of glycolytic/tricarboxylic acid-cycle intermediates, adenine nucleotide pools and the cellular redox-status in the infective (L3) larvae of (homogonic) Strongyloides ratti.

Infective (L3) larvae of Strongyloides ratti (homogonic strain) were freeze-clamped (-196 degrees C) and the steady-state content of the glycolytic, Krebs tricarboxylic acid (KTA)-cycle intermediates and adenine nucleotides analysed. Comparison of the mass-action ratios (MARs) of the glycolytic enzymes with their apparent equilibrium constants (K9eq) indicate that phosphoglucomutase, glucosephosphate isomerase, triosephosphate isomerase, phosphoglyceromutase and phosphopyruvate hydratase reactions were all at or near equilibrium, whilst hexokinase, phosphofructokinase and pyruvate kinase were displaced from equilibrium. The S. ratti aldolase and myokinase appear to be somewhat displaced from equilibrium and thus may have pseudoregulatory roles. The adenylate energy charge (AEC), ATP/ADP ratio and the available adenylate energy (AAE) indices were 0.9 +/- 0.04, 8.76 +/- 1.5 and 397 +/- 43, respectively. The free [NAD+]/[NADH+H+] ratio of the cytoplasmic compartment of S. ratti L3 larvae calculated employing the steady-state content of the oxidised and reduced substrates of lactate dehydrogenase (E.C. 1.1.1.27) and the combined glyceraldehyde 3-phosphate dehydrogenase (E.C. 1.2.1.12)/3-phosphoglycerate kinase (E.C. 2.7.2.3) system were ca. 523 and 1200, respectively. The free[NAD+]/[NADH+H+] ratio in the mitochondrial compartment of S. ratti L3 larvae calculated using the malate dehydrogenase (E.C. 1.1.1.37) equilibrium was found to be 1962:1. The data is discussed with respect to the predominantly aerobic nature of the energy metabolism of the L3 larvae.

Adenine Nucleotides↗

Strongyloides stercoralis: loss of ability to disseminate after repeated passage in laboratory beagles.

A strain of Strongyloides stercoralis originally isolated from a south-east Asian patient and maintained by passage in monkeys for four years was introduced into parasite-free laboratory-reared beagles. After the second passage in these dogs, the prepatent period shortened from an average of 12-14 d to 8-9 d. Up to the third generation, immunosuppressive treatment of the dogs with oral corticosteroids caused hyperinfection with dissemination, albeit limited, to extra-intestinal organs. From the fourth generation onward, neither hyperinfection nor extra-intestinal migration could be induced even by higher doses of corticosteroids. These observations suggest that the biological behaviour of populations of S. stercoralis can alter as they pass from host to host. Such changes may account, at least in part, for the diverse clinical manifestations of strongyloidiasis.

Animals↗

Immunodiagnosis of Strongyloides stercoralis infection: a method for increasing the specificity of the indirect ELISA.

Indirect enzyme-linked immunosorbent assay (ELISA) allows sensitive detection of serum immunoglobulin (Ig) G against a soluble extract of Strongyloides stercoralis infective larvae. In this study, 40/40 (100%) human strongyloidiasis sera had high levels of anti-S. stercoralis IgG, but 30/40 (75%) filariasis sera, and 12/40 (30%) necatoriasis sera also had higher levels than control sera from UK residents. In attempts to increase the assay specificity by absorption of cross-reactive IgG, the effectiveness of pre-incubation of sera with extracts of different parasitic nematodes was investigated. One hour of incubation with 20 micrograms/ml aqueous extract of Onchocerca gutturosa absorbed cross-reactive IgG in most filariasis and necatoriasis sera, reducing the proportion with IgG levels above the positivity threshold by more than one-half. Preliminary results suggest that absorption with extracts of other filarial nematodes is equally effective, and that some of the cross-reactive IgG is directed against phosphorylcholine. Cross-reactive IgG in most necatoriasis sera was effectively absorbed with 20 micrograms/ml extract of Necator americanus. Cross-reactive IgG was not effectively absorbed with an extract of Ascaris lumbricoides. Absorption of cross-reactive IgG is an effective means of increasing the specificity of the indirect ELISA, for use in the immunodiagnosis and immuno-epidemiology of S. stercoralis infection.

Absorption↗

Treatment of Strongyloides stercoralis infection with ivermectin compared with albendazole: results of an open study of 60 cases.

Ivermectin is highly effective against animal intestinal nematodes and is used in the treatment of onchocerciasis in humans. A study was undertaken to compare the efficacy of the drug with that of albendazole in the treatment of uncomplicated strongyloidiasis. Sixty patients with confirmed Strongyloides stercoralis infection were enrolled in an open randomized study and given either albendazole, 400 mg/d for 3 d or ivermectin, 150-200 micrograms/kg in a single dose. Efficacy and tolerance were evaluated on days 7, 30 and 90. Each visit included a parasitological examination of 3 stool specimens, using saline and Kato smears and formalin-ether and Baermann concentrations. Fifty-three patients were eligible for evaluation. Parasitological cure was obtained in 24 of the 29 patients treated with ivermectin (83%) and in 9 of the 24 patients who were given albendazole (38%); ivermectin was significantly more effective than albendazole (P < 0.01). Clinical and biological adverse reactions were negligible in both treatment groups. The 20 patients who failed therapy were given a second treatment course with 150-200 micrograms/kg of ivermectin in a single dose or on 2 consecutive days. Sixteen patients were cured and the other 4 had only incomplete follow-up. Ivermectin therefore constitutes an acceptable therapeutic alternative for uncomplicated strongyloidiasis.

Adolescent↗

Strongyloides stercoralis in an urban slum community in Bangladesh: factors independently associated with infection.

Stool samples from 880 residents in an urban slum in Dhaka, Bangladesh, were collected on 3 occasions over one year, and examined for intestinal parasites. Information on many potential risk factors for infection was obtained by questionnaire from a respondent in each household studied. In a crude univariate analysis of the data, several of the factors were found to be significantly associated with Strongyloides stercoralis infection. Most of these factors were co-variate with one another, and with poverty generally. Using Mantel-Haenszel chi 2 tests to control for confounding effects of each variable individually, the following 4 factors remained independently associated with S. stercoralis infection: respondent's use of a community latrine rather than a private latrine, living in a house with an earth floor rather than a cement floor, being of Bihari ethnicity, and being 7-10 years of age. Implications of these results for the epidemiology and control of strongyloidiasis are briefly discussed.

Adolescent↗

Household aggregation of Strongyloides stercoralis infection in Bangladesh.

Strongyloides stercoralis infections were shown to be aggregated in households in an urban slum community in Dhaka, Bangladesh. Parasitological data on 880 residents living in 280 households were analysed statistically using 3 different tests, each of which yielded significant evidence of household aggregation of S. stercoralis infection. One test was applied to the data after stratification for 4 variables were previously shown to be independently associated with infection. Evidence of household aggregation of infection remained after stratification, suggesting that aggregation is due not only to shared risk factors, but also to either familial genetic predisposition to infection or close contact person to person transmission of infection within households.

Adolescent↗

Antigens from the surface and excretions/secretions of the filariform larva of Strongyloides stercoralis.

The surface and excretory/secretory (ES) antigens of the infective, filariform larva (L3) of Strongyloides stercoralis were identified. These studies provide a basis for the purification of these proteins as diagnostic allergens for human strongyloidiasis. The Mr values of the surface and ES molecules were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography, fluorography, or silver staining following the recovery of these molecules after the radiolabelling of living parasites. At least 10 surface proteins were radioiodinated extrinsically using chloroglycoluril as the catalyst for iodination, and then extracted with detergents and/or beta-mercaptoethanol. Several surface molecules of the L3 were immunogenic in humans, as determined by immunoprecipitation with sera (IHS) from infected patients. About 30 proteins were present in the ES preparation. Many ES antigens were labelled biosynthetically during the culture of larvae in media supplemented with either [35S]methionine or [14C]glucose. Furthermore, several of the surface proteins of the L3 were found with the ES antigens recoverable by culturing larvae in vitro. About 10 of the ES proteins were immunogenic as determined by immunoaffinity chromatography using IHS; and two of these antigens with Mr 50,000 and 90,000 incorporated [35S]methionine during culture of larvae. Moreover, some ES proteins were allergenic when tested in an in vitro assay of histamine release from basophils from infected humans or monkeys. The isotype of the homocytophilic antibodies involved in this immediate hypersensitivity assay, which is the basis of a diagnostic skin test for human strongyloidiasis, appears to be IgE.

Animals↗

[Strongyloides stercoralis in T-cell leukemia/lymphoma in adults and acquired immunodeficiency syndrome].

The mechanisms of chronic infestation by Strongyloides stercoralis (Ss) are unknown. Immunodepression is classically evoked to explain the proliferation of the parasite that is sometimes massive and overwhelming. We present here two retrospective studies of 13 cases of strongyloidiasis (three disseminated strongyloidiasis) out of 26 patients with adult T-cell leukemia/lymphoma (ATL) and of ten cases of strongyloidiasis out of 98 patients with AIDS. Ten patients out of 98 were dually infected with both HIV and HTLV-1: 2/10 also infected by Ss. The frequency of Ss infection appears higher in ATL patients when compared with AIDS patients (P < 0.001). Ss-infected ATL patients were younger than those uninfected (P < 0.01). Ss-infected AIDS patients were older than the ones uninfected (p < 0.03). No significative difference was found between ATL and AIDS patients who were over 40 years-old. These data suggest that 1) the particular type of immunodepression produced early by HTLV-1 is more favorable to the development of Ss infection than the one associated with HIV, 2) the latency of expression of HTLV-1, prior to the development of leukemia, is reduced in Ss-infected patients, 3) Ss infection may slow down VIH pathogenic activity.

AIDS-Related Opportunistic Infections↗

Solubilization studies on the epicuticular antigens of Strongyloides ratti.

Antigens on the epicuticular surface of Strongyloides ratti infective third-stage larvae (L3) were demonstrated by both ferritin-conjugated antibody and indirect fluorescent antibody techniques. The rat antibodies from immune serum that bind to these antigens were chiefly of the IgG2a subclass. Solubilization of these antigens by extraction with detergents, hypertonic salt, organic solvents and by freezing and thawing was limited as measured by the reduction in antibody binding to the epicuticle. The epicuticular antigens were resistant in situ to degradation by a variety of proteases, carbohydrases and lipase. Infectivity of the L3 in the rat was not reduced by prior sensitization with rat antibody. The epicuticular antigens are not completely species-specific since antibody from S. ransomi-infected pigs cross-reacted well with S. ratti L3 antibody. However, high levels of resistance to S. ratti could be induced in rats only by multiple inoculations of heat-killed S. ratti L3.

Animals↗

The development of the free-living stages of Strongyloides papillosus. I. Effect of temperature on the development of the heterogonic and homogonic nematodes in faecal culture.

Temperature had a significant effect on the proportion of free-living females and infective larvae of Strongyloides papillosus developing from faecal culture. The number of free-living males recovered from cultures incubated at 20 degrees C and 30 degrees C was fairly constant. However, females were most numerous at 30 degrees C and infective larvae at 20 degrees C, with the sum of the two approximately the same at both temperatures. This indicates that infective larvae and females are developing from genetically identical eggs. The critical period by which the course of development has been fixed occurs between 4 and 5 h at 30 degrees C, but between 4 and 24 h at 20 degrees C.

Animals↗

Sudden death in calves associated with Strongyloides papillosus infection.

In recent years, a total of 152 calves have died suddenly on three farms in the southern part of Japan which used sawdust as litter. Calves often died within a few minutes of a violent fall without showing previous clinical signs. On necropsy of these cases, only a slight lesion was observed, although all showed heavy infection with Strongyloides papillosus (SPL). They appeared to have not only a large number of SPL eggs in the faeces, together with many adult SPL worms in the intestine, but also many migratory SPL larvae in tissues such as the lung and muscle. These outbreaks ceased following medication with thiabendazole or ivermectin. It is concluded that 'sudden death' is associated with heavy infection with SPL.

Animal Husbandry↗

Dynamics of faecal egg counts in rabbits experimentally infected with Strongyloides papillosus.

An experiment using rabbits was carried out to determine basic data to maintain the calf strain of Strongyloides papillosus (SPL). Rabbits treated with four different numbers of doses of dexamethasone were exposed to 50,000 or about 100,000 infective larvae of SPL (Himeji strain, 1988). As a result, faecal egg output was valid in individual rabbits, regardless of dexamethasone administration. Eggs appeared in the faeces 9 or 10 days after exposure, and increased to as much as 20,000-573,000 eggs per gram (EPG). Thirteen of 18 rabbits died (72%) 12-29 days after infection. Five surviving rabbits discharged eggs in their faeces for more than 1 month; however, the EPG value of these cases on the last day was comparatively low.

Animals↗

Sudden death of calves by experimental infection with Strongyloides papillosus. I. Parasitological observations.

Recently, an unknown disease, 'sudden death', in calves has been found in Japanese beef production farms. A previous study conducted by Taira and Ura indicated that sudden death can be effected in calves by hyperinfection of Strongyloides papillosus (SPL) and that the disease is possibly caused by SPL infection. In the present work, an experimental infection of SPL in calves was conducted to confirm the field occurrence. Fifteen Holstein Friesian calves, ranging from 45.5 to 85.6 kg in body weight, were divided into six groups. Calves of Groups A, B, C, D, E and F were infected once at the rate of 100,000, 320,000, 1,000,000, 3,200,000, 10,000,000 and 32,000,000 SPL larvae per 100 kg of body weight, respectively. Five calves were assigned to Group B, while two calves were assigned to the other groups. After showing no premonitory signs, sudden death of ten calves took place. The survival time of these calves was 27.4 and 16.8 days (Group B), 14.8 and 14.8 days (Group C), 13.3 and 14.2 days (Group D), 11.0 and 11.1 days (Group E) and 11.6 and 10.8 days (Group F). Three calves of Group B did not exhibit sudden death. The results of this study demonstrate that strongyloidiasis was the cause of sudden death.

Animals↗

Natural infections of Strongyloides westeri: prevalence in horse foals on several farms in central Kentucky in 1992.

During the period 28 February-1 July 1992, fecal samples were collected and examined for eggs of Strongyloides westeri once from each of 382 horse foals (364 Thoroughbreds, 16 Standardbreds, one Lippizaner, and one draft-type horse) in central Kentucky. Ages of the foals at the time of sampling ranged from 7 to 63 days (mean 22 days). The nine farms in the study were considered to have overall excellent deworming programs. None of the foals had been treated with an antiparasitic compound before the study. Eggs of S. westeri were found in 6% (22 of 382) of the foals on 78% (7 of 9) of the farms. Current prevalence of this parasite in foals is discussed relative to studies before the availability and usage of effective drugs.

Animals↗

Is there an association between positive Strongyloides stercoralis serology and diabetes mellitus?

The purpose of this study was to determine the frequency of association between positive Strongyloides stercoralis serology and diabetes mellitus. A total of 78 diabetic patients and 42 controls were evaluated. For a parasitological diagnosis, Baermann and Hoffman et al.'s methods were applied. The immunological diagnosis involved the indirect fluorescence antibody test, ELISA and Western blotting to detect IgG antibodies. The frequency of positive S. stercoralis serology in diabetics was 23% versus 7.1% in the control group (P<0.05). The odds ratio for diabetics was 3.9 (CI, 1.6-15.9, P<0.05). Diabetic patients with HbA(1c)< or =7 had a greater chance of testing negatively for S. stercoralis infection (OR: 1.5, P>0.05). Provided there are related cases of disseminated strongyloidiasis in diabetics and there is a higher frequency of asymptomaticity of the infection in this group, the immunological screening of these patients at risk could prevent severe and fatal outcomes of the disease.

Adult↗