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Seroprevalence data illustrate that human exposure to Toxocara is frequent. Environmental contamination with Toxocara spp. eggs is assumed to be the best indicator of human exposure, but increased risk of exposure has also been associated with many other factors. Reported associations are inconsistent, however, and there is still ambiguity regarding the factors driving the onset of Toxocara antibody positivity. The objective of this work was to assess the validity of our current conceptual understanding of the key processes driving human exposure to Toxocara. We constructed an agent-based model predicting Toxocara antibody positivity (as a measure of exposure) in children. Exposure was assumed to depend on the joint probability of 3 parameters: (1) environmental contamination with Toxocara spp. eggs, (2) larvation of these eggs and (3) the age-related contact with these eggs. This joint probability was linked to processes of acquired humoral immunity, influencing the rate of antibody seroreversion. The results of the simulation were validated against published data from 5 different geographical settings. Using simple rules and a stochastic approach with parameter estimates derived from the respective contexts, plausible serological patterns emerged from the model in nearly all settings. Our approach leads to novel insights in the transmission dynamics of Toxocara.

In human Toxocara canis infection, an association has been shown between high IgG avidity in the chronic phase and low IgG avidity in recently acquired toxocarosis. The evolution of the antibody response in terms of avidity has been carried out through a T. canis infection in BALB/c mice. Infection with T. canis embryonated eggs (EE) was carried out with single doses (SD) of 6, 12, 50, 100, 200 or 1000 EE/mouse and with multiple doses (MD) of 200 and 1000 EE. Specific antibodies against T. canis (IgM+G, IgG, IgG1 and IgM) were detected by ELISA and Western Blot (WB) techniques in the presence and absence of urea. With the ELISA method, an increase in the avidity index (AI) of around 50% was detected from days 40-80 p.i. to the end of the study, with all the doses studied. The WB method showed the presence of high avidity antibodies bound to 100 kDa and 75 kDa T. canis proteins in all the cases when the IgM+G and the IgG1 antibodies were investigated. Antibodies of variable avidity were observed in those sera that recognized the group of low molecular weight proteins, between 37 kDa and 25 kDa.

Toxocara canis, the parasitic roundworm of dogs, can infect a number of paratenic hosts, such as mice and humans, due to the widespread dissemination of its ova in the environment. In these paratenic hosts, larvae have been shown to exhibit a predilection for the central nervous system, resulting in an increasing number of parasites migrating to the brain as infection progresses. In an initial experiment, we investigated the differential brain involvement of T. canis in 7 strains of inbred mice, and chose 2 strains, susceptible (BALB/c) and resistant (NIH) to cerebral infection. In a second experiment, both strains were investigated in terms of course of migration, larval accumulation, and behavioural response to T. canis infection. Results revealed that infected BALB/c mice took significantly longer to drink from a water source (following a period of deprivation), compared with control mice, indicating some degree of memory impairment. Cerebral larval recoveries from both strains of mice demonstrated variation between the two experiments, suggesting that larval burdens may not be a reliable indicator of susceptibility or resistance to T. canis infection. The percentage of total recovered larvae in each organ may be a better representation of larval distribution. Our model system may provide insights into the impact of chronic geohelminth infection on cognitive development.

Toxocariasis is a common helminthic infection that has a worldwide distribution. However, data from Lebanon about the prevalence of this infection are non-existent. We conducted a Toxocara seroprevalence study with 150 subjects attending the outpatient clinics at the American University of Beirut Medical Center between May and June 2004. Serum specimens were tested for anti-Toxocara antibodies by enzyme-linked immunosorbent assay and confirmed by Western blot. Multivariate analysis was performed to identify risk factors for infection. The seroprevalence rate of toxocariasis was 19%. Male gender and below high school education were significantly associated with a positive serological test (odds ratios = 3.1 and 2.8, respectively). Higher numbers of persons in the household, and low family income during childhood, were significant on bivariate analysis only. Toxocariasis is common in Lebanon. A large population-based survey is needed to confirm these results.

The impact of Toxocara canis infection of Balb/c mice mothers on the future immune response of their offspring towards reinfection with the same parasite was studied. Two groups of offspring, the first originating from the mothers infected with a single dose of 1000 Toxocara canis eggs and the second from non-infected mothers, were both challenged with 500 T. canis eggs per animal at 6 weeks of age. The proportions of spleen CD4+ and CD8+ T lymphocytes, the level of serum cytokines IFN-gamma and IL-5, eosinophilia in peripheral blood, the production of specific antibodies and the number of migrating larvae were monitored. In both groups of offspring, the challenge infection resulted in an increase in CD4+ T-cell subtype in comparison with the non-infected healthy control, although after an initial decline a subsequent increase in CD8+ was observed. The immunoregulation index (CD4+/CD8+) was lower in the group of mice originating from infected mothers throughout the whole experiment compared to the offspring of non-infected mothers as well as in healthy control mice of the same age. In the offspring of infected mothers mainly, a reduced production of IFN-gamma and of IL-5, suppressed eosinophilia and a higher level of protective antibodies was detected, compared to the control second group, in which the INF-gamma concentration significantly increased after day 42 p.i. In the first group of offspring before challenge, 12.7 +/- 2.5 larvae in the brains and 32 +/- 2.1 larvae in the muscles transmitted from the infected mothers were detected. There was a significant reduction in larval recovery from brain on days 42 and 49 p.i. (56.7 and 56.8%, respectively), while from muscles in the same time there was a reduction of 46.7 and 39%, respectively, compared to the offspring of non-infected mothers. These results indicate a significant protective memory of immune mechanisms against T. canis induced in offspring of Toxocara-infected mother mice.

The population dynamics of Toxocara canis in pigs, and their immune response to a primary and a challenge infection, were studied by parasitological, haematological and serological parameters. Seventy pigs were divided into 4 groups; 35 pigs received a primary infection (group A), 15 pigs received both a primary and a challenge infection (group B), 15 pigs received the challenge infection only (group C), and 5 pigs served as helminth-free controls (group NC). A dose of 50,000 eggs was administered for the primary infection (day 0) and a dose of 10,000 eggs was given for the challenge infection (day 28). On days 7, 14, 21 and 28 p.i., 5 pigs of group A, and on days 35, 42 and 49 p.i., 5 pigs from each of groups A, B and C were necropsied. Numbers of recovered larvae varied widely among the 5 pigs of each group on all days of necropsy. Toxocara canis larvae were recovered predominantly from the lungs; migration of larvae to other organs or tissues from the lungs was restricted. In group A, the larval burden in the lungs peaked on day 14 p.i., and the larval densities decreased significantly over time. Thereafter, the majority of larvae were recovered from the lungs until the end of the experiment (day 49 p.i.). A few larvae were found in the muscles and brain until day 42 p.i., and 2 larvae were found in the eyes of 2 pigs on day 35 p.i. There was little evidence of protective immunity to a challenge infection in this experiment. The eosinophil levels tended not to increase in pigs receiving a challenge infection, in contrast to the challenge control pigs. The fact that T. canis larvae migrate and persist in the tissues of pigs for more than 1 month suggests a zoonotic risk in infected pigs. The relevance of these data to the population biology and immunology of porcine and human toxocarosis is discussed.

Infective larvae of the dog roundworm Toxocara canis survive in the tissues of their hosts for extended periods in a state of developmental arrest, successfully evading immune destruction. This survival strategy is thought to be mediated by T. canis excretory/secretory (TES) products which downregulate or divert the immune response. We purified one of the major TES products, TES-70 and gained amino acid sequence from 4 tryptic peptides. These peptides were matched to a predicted protein from a cDNA that was isolated by expression screening a T. canis cDNA library with mouse anti-TES serum. The predicted protein (Tc-CTL-4) is similar to, but larger than, Tc-CTL-1, a 32-kDa C-type lectin secreted by T. canis larvae. Tc-CTL-4 has a signal peptide, 2 Cys-rich domains and a C-terminal calcium-dependent C-type lectin domain that shares sequence similarity with host immune cell receptors such as macrophage mannose receptor and CD23. The lectin domain was expressed in bacteria and antiserum to the purified recombinant protein was used to confirm that Tc-ctl-4 did encode the native TES-70 glycoprotein. TES-70 selectively bound to ligands on the surface of Madin-Darby Canine Kidney cells in vitro in a calcium-dependent manner, inhibitable by mammalian serum, indicating that a host glycan is the native ligand for this new parasite lectin.

An open randomized study was carried out to assess the efficacy of diethylcarbamazine (DEC) and mebendazole (MBZ) across, respectively, 39 and 41 patients, all exhibiting clinical and biological features evocative of toxocariasis, together with a positive immunodiagnosis by Western blot using Toxocara larvae excretory-secretory antigens. Clinical impact of the disease was quantified by a score system (20 parameters). Post-treatment follow-up used also results from eosinophil counts from total and specific anti-Toxocara IgE quantitations, and from Western blots. All patients attended the control check-up 1 month after the end of treatment. Analysis of the results showed a similar good efficacy of DEC and MBZ on clinical score and eosinophil count that were significantly lowered. MBZ therapy was slightly more efficient on specific anti-Toxocara IgE kinetics, while total IgE mean titres were not affected by either DEC or MBZ treatment. Patients from the DEC group reported a significantly higher rate of adverse reactions, most of which could be due to parasite lysis. This fact could be a strong argument for treating human toxocariasis with MBZ.

Antibodies to Toxocara were measured by enzyme-linked immunosorbent assay (ELISA) in sera of ascarid-free dogs which had been fed 0, 100 or 10 000 embryonated T. canis eggs. Dogs fed 10 000 eggs developed higher titres than those given 100 eggs and larvae failed to complete their migration to the gut. In addition, in dogs given 100 eggs, there was a significant negative correlation between the number of worms in the gut and granulomatous foci in the lung. The data suggest that the ELISA is sensitive and specific for Toxocara infection in dogs.

In this study, we have developed a reliable radioimmunoassay (RIA) for Ascaris suum protein (Asp) and have measured the serum concentration of Asp in control subjects and in patients with helminthiasis and a variety of gastrointestinal diseases. The cross-reactivities of Asp antibody with other helminth worm antigens were also measured by RIA. Ascaris lumbricoides contained approximately the same amount of Asp antigenic substance as did Ascaris suum. Toxocara canis had a high concentration of a substance partially cross-reactive with Asp. Small amounts of substance cross-reacting with Asp were also exhibited by Anisakis larvae, Schistosoma japonicum and Taenia saginata. However, no cross-reactivities of Fasciola hepatica and Paragonimus westermani with Asp were observed in this study. Almost all the serum levels of Asp from normal control subjects were below the sensitivity of this RIA (10 ng/ml). High concentrations of Asp were observed in sera from patients with ascariasis (64.5 +/- 18.8 ng/ml: mean +/- S.E.), anisakiasis (75.2 +/- 28.0 ng/ml), toxocariasis (78.4 +/- 31.3 ng/ml) and schistosomiasis japonica (14.3 +/- 3.4 ng/ml). Asp levels in sera from patients with other helminthiasis were not significantly high. From these findings, it was suggested that RIA for Asp may not be specific for diagnosis of ascariasis, but might be a suitable screening method for patients with some kinds of helminthiasis. Interestingly, high concentrations of Asp were also observed in sera from patients with ulcerative colitis (24.8 +/- 2.5 ng/ml) and Crohn's disease (19.8 +/- 5.2 ng/ml), but Asp levels from 7 other gastrointestinal diseases seemed not to be significantly high. These findings remain unexplained but may derive from cross-reactivities between helminthic and gut mucosal antigens.