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Observations on the transmission and development of Toxocara pteropodis (Ascaridoidea: Nematoda) in the Australian grey-headed flying-fox, Pteropus poliocephalus (Pteropodidae: Megachiroptera).

Findings in the Australian Grey-Headed Flying-Fox, Pteropus poliocephalus, have elucidated the life-cycle of Toxocara pteropodis. In adult bats, other than parturient females, larvae were found only in the livers. Following parturition, larvae were recovered only from mammary glands up to 2 weeks post-partum. Developing larvae were found only in the intestine of young bats from the age of two days onwards; there was no evidence of pulmonary migration. The evidence indicates that juvenile bats commence passing Toxocara eggs in their faeces at about 2 months of age and expel the worms spontaneously following weaning at about 5 months. The eggs passed in the faeces of the young bat and its mother are disseminated throughout their environment and embryonate rapidly, being infective to mice after 10 days. Under natural conditions the eggs remain viable for 6 weeks or less and are infective to bats by the oral route.

Animals↗

Toxocara canis: interaction of human blood eosinophils with the infective larvae.

This study was carried out to investigate the nature of the immunological responses which took place in a child who had recently recovered from toxocariasis. She had developed a marked eosinophilia and had high titers of toxocara antibodies. Experiments were performed to examine whether Toxocara canis infective larvae could be killed in the presence of her serum and human eosinophils. Eosinophils with human complement, or this patient's serum, adhered to the surface of the larvae within 10 min. By 40 min, using both light and electron microscopy, it was shown that the cells had flattened against the cuticle and degranulated. However, by 3 hr, eosinophils had begun to detach, and the larvae remained alive for at least 1 week afterward. Further addition of serum or of eosinophils, which were shown to be able to immobilize T. spiralis infective larvae, failed to kill the T. canis larvae. It was concluded that, in this patient, the development of an inflammatory response to a T. canis infection was not associated with the appearance of antibodies capable of inducing eosinophil dependent toxicity to the larvae in vitro. Eosinophil dependent killing mechanisms may be less important than other components of the immune response, in immunity to this parasite in humans.

Animals↗

Toxocara canis: monoclonal antibodies to larval excretory-secretory antigens that bind with genus and species specificity to the cuticular surface of infective larvae.

When maintained in culture, the infective-stage larvae of Toxocara canis produce a group of excretory-secretory antigens. Monoclonal antibodies to these antigens have been produced and partially characterized. Hybridomas were made using spleens from mice that had been given 250 embryonated eggs of T. canis followed by immunization with excretory-secretory antigens. Monoclonal antibodies were first screened against excretory-secretory antigens using an indirect enzyme-linked immunosorbent assay. Those antibodies positive in this assay were then screened against the surfaces of formalin-fixed, infective-stage larvae using an indirect fluorescent antibody assay. The two monoclonal antibodies showing fluorescence were also tested against the surfaces of infective-stage larvae of Toxocara cati, Baylisascaris procyonis, Toxascaris leonina, Ascaris suum, a Porrocaecum sp., and Dirofilaria immitis. One of these two antibodies bound to the surface of T. canis and T. cati while the other bound only to the surface of T. canis; neither were reactive with the other ascaridoid larvae or the larvae of D. immitis. Enzyme-linked immunoelectrotransfer blotting techniques were used to demonstrate that the cross-reactive antibody recognized antigens with molecular weights of about 200 kDa while the more specific monoclonal antibody recognized antigens with approximate molecular weights of 80 kDa. The specificity of these two antibodies for T. canis and T. cati should prove helpful in the development of more specific assays for the diagnosis of visceral and ocular larva migrans.

Animals↗

Toxocara canis: a labile antigenic surface coat overlying the epicuticle of infective larvae.

An electron-dense coat covering the surface of Toxocara canis infective-stage larvae is described. This coat readily binds to cationized ferritin and ruthenium red, indicating a net negative charge and mucopolysaccharide content, and can be visualized by immuno-electron microscopy only if cryosectioning is employed. Monoclonal antibodies reactive to the surface of live larvae bind the surface coat but not the underlying cuticle in ultrathin cryosections. The surface coat is dissipated on exposure to ethanol, explaining the lack of surface reactivity of conventionally prepared immunoelectron microscopy sections of T. canis. Differential ethanol extraction of surface-iodinated larvae demonstrates that the major component associated with the coat is TES-120, a 120-kDa glycoprotein previously identified by surface iodination, which is also a dominant secreted product. The surface-labeled TES-70 glycoprotein is linked with a more hydrophobic stratum at the surface, while a prominent 32-kDa glycoprotein, TES-32, is more strongly represented within the cuticle itself. Antibody binding to the coat under physiological conditions results in the loss of the surface coat, but this process is arrested at 4 degrees C. This result gives a physical basis to earlier observations on the shedding of surface-bound antibodies by this parasite. An extracuticular surface coat has been demonstrated on Toxocara larvae prior to hatching from the egg and during all stages of in vitro culture, suggesting that it may play a role both in protecting the parasite on hatching in the gastrointestinal tract and on subsequent tissue invasion in evading host immune responses directed at surface antigens.

Animals↗

Toxocara canis larvae in the brain of a British child.

The clinical and autopsy findings of a two and a half year-old infant with Toxocara sp. infection of the brain and granulomatous lesions in the liver are reported. The cause of death was non-accidental injury. The relationship between Toxocara infection and behavioural disorders is discussed.

Ascariasis↗

Infection of Japanese quail with Toxocara canis larvae and establishment of patent infection in pups.

Migration and distribution of Toxocara canis larvae in the tissues of Japanese quail, infected orally with 5 X 10(3) infective eggs, and the establishment of patent infection from the quails to the definitive host, were studied. Larval yield at necropsy from various tissues and organs of quail varied from 4.72 to 7.54% of the infective eggs inoculated within the period 1-60 days post-infection (PI). The total number of larvae recovered on different days showed a gradual increase. The percent inoculum recovered at necropsy was highest on Day 60. Most of the larvae were found in the liver throughout the period and only a few migrated to other tissues, such as lung, heart, muscle and brain. The establishment of patent infections in the definitive host was studied by feeding the 15-day infected livers of Japanese quail (400 larvae) to Toxocara-free pups. Eggs first appeared in the faeces 38 days post-infection, the mean worm burden at necropsy was 87 and the percentage of infection established was 21.75%. Thus the role of Japanese quail as a paratenic host of T. canis is established.

Animals↗

Influence of soil texture in the recovery of Toxocara canis eggs by a flotation method.

In epidemiological surveys, the evaluation of soil contamination by Toxocara canis eggs requires a quick and easy method for the isolation of parasite eggs from soil samples. The efficiency of flotation methods is influenced by sample size, soil texture, degree of soil contamination, pretreatment, flotation solutions and time of flotation. This investigation was designed to evaluate the influence of soil texture in the recovery of T. canis eggs with the centrifugal flotation technique of Dada (Dada, B.J.O., 1979. A new technique for the recovery of Toxocara eggs from soil. J. Helminthol., 53: 141-144). Four types of soil (clay silt, sandy, silty clay and sand) were artificially contaminated with T. canis eggs (200 eggs per gram). Zinc sulphate (specific gravity 1.20) and sodium dichromate (specific gravity 1.35) were used as flotation solutions. Twenty replicated examinations were performed for each type of soil and flotation solution. There was a statistically significant difference in the results depending on soil type. The highest recovery percentages were observed in soils rich in sand (62.5% for sand and 38.0% for sandy soil). Differences were also observed with different flotation solutions. Sodium dichromate solution was more efficient for recovering T. canis eggs, regardless of the soil texture.

Analysis of Variance↗

Cytosolic malate dehydrogenase in muscle extracts of Toxocara canis.

1. Malate dehydrogenase (L-malic acid:NAD+ oxydoreductase, EC 1.1.1.37) was partially purified from muscle extracts of Toxocara canis by means of gel chromatography in Sephadex G-150 and affinity chromatography in Sepharose-4B-Blue dextran. 2. The purified enzyme was very active in reducing oxalacetate and less active in oxidizing L-malate. It was inhibited by excess oxalacetate but not by L-malate. 3. The kinetic parameters of the enzyme were obtained and these included: pH and temperature optima and apparent Michaelis constants for the substrates. 4. The results suggest that the enzyme from Toxocara canis behaves like the enzyme of the model helminth Ascaris lumbricoides.

Animals↗

Purification and properties of mitochondrial malate dehydrogenase of Toxocara canis muscle.

Mitochondrial malate dehydrogenase was purified from muscle extracts of Toxocara canis by means of Sephadex G-100 gel filtration, DEAE-Sephadex ion-exchange chromatography and 5'AMP-Sepharose 4B affinity chromatography. The purified enzyme showed an optimum pH for the reduction of oxaloacetate of 7.3 in Tris-HCl buffer and of pH 7.5-7.8 in phosphate buffer. The m-MDH showed values of 3.2 kcal/mol and 10.5 kcal/mol for the energy of activation, calculated from the Arrhenius equation. The mitochondrial enzyme was found to be more susceptible to thermal inactivation as compared with the cytosolic isoenzyme. Kinetic experiments showed that the m-MDH of Toxocara canis is inhibited by excess oxaloacetate but not by excess NADH. The apparent Km for oxaloacetate reduction was 53 microM and 0.54 mM for L-malate oxidation.

Animals↗

Toxocara canis: effect of inoculum size on pulmonary pathology and cytokine expression in BALB/c mice.

Infection of mice with Toxocara canis results in pulmonary inflammation and the induction of a Th2 type of immune response. The aim of this study was to determine whether the effect of infection with this nematode depends on the inoculum size. Results indicate that mice infected with either a high or a low inoculum size showed, in a dose-dependent manner, pulmonary inflammation with eosinophil infiltration, increased levels of total IgE, and Toxocara-specific IgG1 that persisted up to 60 days p.i. Relative quantification of cytokine expression in lungs of mice infected with different doses showed proportional increased expression of the IL-4, IL-5, and IL-10 transcripts, whereas the expression of the IFN-gamma transcript was not different from that of uninfected controls. These results indicate that infection of BALB/c mice with T. canis results in chronic pulmonary inflammation and a dominant Th2 type of immune response, independent of the inoculum size.

Animals↗

A putative role for Toxocara species in the aetiology of multiple sclerosis.

Multiple sclerosis (MS) is a disease of unknown aetiology. The finding of monoclonal antibodies in MS has been attributed to various infectious agents. Nematodes, such as Toxocara species have not been explored as possible aetiologic agents of MS. Some epidemiological studies have found an association between exposure to stress and household pets prior to the diagnosis of MS. In a case known to the authors, slight malaise and eosinophilia in peripheral blood preceded the diagnosis of MS by one year in a middle-aged man who lived in rural surroundings with cats in the household. The ubiquitary parasite Toxocara catis or canis is prevalent and serum antibodies are found regularly in populations examined. It is able to develop into the larval stage in human beings. The hypothesis presented here is that MS could be initiated by such infections in previously unexposed subjects under conditions of long-term stress.

Animals↗

Zoonotic risk of Toxocara canis infection through consumption of pig or poultry viscera.

The potential zoonotic risk of Toxocara canis infections from consumption of swine or poultry viscera containing larvae was assessed using a pig model. Two groups of six pigs were fed either fresh swine viscera (group FS) or poultry viscera (FP) containing around 3500 Toxocara larvae. Another two groups of six pigs were fed swine viscera (PS) or poultry viscera (PP) preserved at 4 degrees C for 1 week. All pigs were necropsied 14 days after the exposure. Liver white spots were counted and T. canis specific IgG antibodies were measured by ELISA. Larval burdens were assessed in the mesenteric lymph nodes, liver, lungs, brain, tongue, and eyes. All recipient pigs exhibited several white spots on the liver surface and detectable antibody levels. Larvae were recovered predominantly from the lungs, but also from the mesenteric lymph nodes and the liver, a few larvae were found in the brain and tongue of the pigs. Two larvae were found in the eyes of two pigs in group FS. Mean percentages of total larval recoveries in groups FS, FP, PS, and PP were 75.3, 63.6, 42.6, and 18.8%, respectively. Significantly higher numbers of larvae were recovered from pigs given swine viscera than pigs given poultry viscera. The preservation at 4 degrees C for 1 week caused a significant reduction in the larval infectivity overall, nevertheless, the recoveries remained substantial. The fact that larvae migrating in swine or poultry organs and tissues have high infectivity in pigs even after preservation at 4 degrees C for 1 week, suggests that human infection with T. canis might easily occur following consumption of raw or undercooked dishes, either fresh or refrigerated, prepared from swine or poultry organs and tissues harbouring T. canis larvae.

Animal Feed↗

Prevalence of Toxocara cati and other parasites in cats' faeces collected from the open spaces of public institutions: Buenos Aires, Argentina.

Toxocarosis is a worldwide parasitic infection that affects both cats and dogs. Toxocara cati (Schrank, 1788) syn. Toxocara mystax (Zeder, 1800) prevalence was studied in faeces from stray cats collected from the open spaces of public institutions of Buenos Aires city, both building and surrounding open spaces are fenced off. Of the 465 samples obtained from March to June of 2005, 58.3% were found to have parasite eggs. The following parasites were identified from the 271 positive samples: T. cati (61.2%), Cystoisospora spp. (20.3%), Trichuris spp. (17.0%), Toxascaris leonina (15.1%), Ancylostoma spp. (14%) and Aelurostrongylus abstrusus (2.6%). T. cati prevalence was 35.7% (95% confidence interval: 31.2-40.1), with a 42.2% single isolations. The most frequent combination was T. cati and Cystoisospora spp. (9%). More than half the areas studied showed over 40% prevalence. Seventy-one percent of the collected samples were fresh with a variable moist consistency and 29% were older with a dry consistency. A statistically significant association was found between sample consistency and presence of parasites (chi2 = 10.81; p = 0.001) as also between sample consistency and presence of T. cati (chi2 = 11.27; p = 0.0007). Moist consistencies were significantly different from the rest: consistency (wet or dry) versus parasites (z = 1.95; p = 0.02) (95% confidence interval: 0.004-0.203); consistency (wet or dry) versus T. cati (z = 3.25; p = 0.0006) (95% confidence interval: 0.075-0.254). The cat population that inhabits these public green spaces contaminates the environment, thus transforming them into dangerous spaces with a variable rate for the human population that spends time in these places.

Animals↗

Contamination of soil with Toxocara eggs in urban (Prague) and rural areas in the Czech Republic.

Contamination of soil with feline and canine ascarid eggs in public parks, backyards and sand pits in Prague, Czech Republic was investigated in this work. Soil samples from shelters and rural areas were also collected. The comparison of soil from different areas (urban, rural, backyards and shelters) exhibited significant difference (chi(2)=32.16, d.f.=3 and p<0.0001). The highest rate of contamination (45%) was found in backyards inhabited by feral cats. The eggs of Toxocara spp. were found in 20.4% of parks, 10% of shelters and 5% of rural samples. Mean egg density per sample from Prague parks was 6.2 eggs/100g of soil. In 126 composite samples from children's and pits, the prevalence of Toxocara eggs was 11.90%. The number of eggs in positive samples varied from 2 to 22 (per 100g). A high proportion (46.9%) of eggs was fully embryonated. There was no difference between the sand pits with or without formal exclusion of dogs (chi(2)=0.6, d.f.=1 and p<0.0001).

Animals↗

Optical coherence tomographic and angiographic findings of a case with subretinal toxocara granuloma.

PURPOSE: To study a case with a subretinal Toxocara granuloma. DESIGN: Interventional case report. METHODS: A patient with a exudative macular lesion, diagnosed as ocular toxocariasis, was examined with optical coherence tomography (OCT) and angiography before and after systemic corticosteroid and anthelminthic therapy. RESULTS: In OCT images, the macular granuloma initially appeared as a highly reflective mass protruding above the retinal pigment epithelium and showed dye leakage by angiography. Posttherapy, the lesion was no longer exudative, was less elevated, and was covered by the retinal pigment epithelium. There was reticular hyperfluorescence surrounded by a hypofluorescent rim in the angiograms. CONCLUSIONS: Subretinal Toxocara granuloma may have a presentation similar to idiopathic choroidal neovascularization (CNV) and should be included in the differential diagnosis of the idiopathic CNV.

Adult↗

Visceral larva migrans: migratory pattern of Toxocara canis in pigs.

The migratory pattern of Toxocara canis was investigated following infection of pigs with 60000 infective eggs. Groups of six pigs were slaughtered at 7, 14 and 28 days after infection (p.i.), and the number of larvae in selected organs and muscles was determined by digestion. A group of uninfected pigs was used as negative controls for blood parameters and weight gain. Toxocara canis migrated well in the pig, although the relative numbers of larvae recovered decreased significantly during the experiment. On day 7 p.i., high numbers of larvae were recovered from the lymph nodes around the small intestine and to some extent also from the lymph nodes around the large intestine, and from the lungs and the liver. On day 14, the majority of larvae were recovered from the lungs and the lymph nodes around the small intestine, and by day 28 p.i. most larvae were found in the lungs. Larvae were recovered from the brain on days 14 and 21, with a maximum on day 14 p.i. No larvae were found in the eyes. Severe pathological changes were observed in the liver and lungs, especially on day 14 p.i.; also, development of granulomas was observed in the kidneys. Finally, a strong specific antibody response towards T. canis L2/L3 ES products was observed from day 14 p.i. until termination of the experiment, and the maximum eosinophil response was observed 14 days p.i. The pig is a useful non-primate model for human visceral larva migrans, since T. canis migrate well and induce a strong immunological response in the pig. However, the importance of the pig as a paratenic host is probably minor, because of the relatively early death of most of the larvae.

Animals↗

Recovery of intraocular Toxocara canis by pars plana vitrectomy.

Pars plana vitrectomy has been used in cases of ocular toxocariasis to remove vitreous opacities due to chronic inflammation and to relieve vitreo-retinal traction causing macular distortion and retinal detachment. To date, the diagnosis of intraocular Toxocara infection in vitrectomy cases has been presumptive, based on characteristic clinical findings, the presence of eosinophils, and corroborative immunodiagnostic tests. The authors report a case in which a vitrectomy for vitreopapillary traction recovered an intact Toxocara canis organism.

Adolescent↗

Immunological and hematological response in experimental Toxocara canis-infected pigs.

The relationship between the immunological and hematological response to infection was studied in pigs inoculated experimentally with Toxocara canis. Two groups of four pigs were infected with doses of 1000 and 2000 infective eggs, respectively. Two uninfected animals were used as negative controls. Blood samples were collected from each pig once a week. Serological examination by ELISA to determine antibody levels against T. canis L2/L3 excretory-secretory (ES) antigens showed values higher than the positive cut-off point (1:32) for both the infected groups. These values increased from day 7 p.i. and remained high during the experimental period until day 56. Significant differences were recorded for the two inoculating doses (p</=0.01) from day 14 to 42 p.i. Western blots run on day 7 p.i. with serum from the 2000 eggs group showed two polypeptide bands of 70 and 55kDa. These same bands were observed for both infected groups from day 15 p.i. until the end of the experiment. Eosinophil counts were significantly higher (p</=0.05) for both inoculated groups between weeks 2 and 7 p.i. as compared with the control group. No significant changes were observed for other blood parameters. The pig is a useful animal model since T. canis induces an early immunological response and ELISA can detect the presence of these antibodies against T. canis L(2)/L(3) ES antigens in animals inoculated with low doses of Toxocara eggs.

Animals↗