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Cerebellar ataxia due to Toxocara infection in Mongolian gerbils, Meriones unguiculatus.

We assessed the usefulness of gerbils as an experimental model for neurologic toxocarosis. Mongolian gerbils, Meriones unguiculatus, infected with Toxocara canis or Toxocara cati (1000 eggs/gerbil) showed progressive neurologic disorders from 50 days after infection in T. canis-infected gerbils or from 120 days after infection in T. cati-infected gerbils. The incidence of the onset was 6 of the 13 gerbils (49%) in the T. canis-gerbils and 5 of the 7 gerbils (71%) in the T. cati-gerbils. Histopathologically, the cerebellum was the most affected in both groups. We observed loss of Purkinje cells, glial nerve fibers, and nerve sheaths. We also found foci consisting of aggregated macrophages scattered in the white matter of the cerebellum. The affected gerbils showed ataxia and ultimately died of cachexia. Our findings suggest that irreversible neurologic toxocarosis in gerbils can be induced by infection with either T. canis or T. cati.

Animals↗

An estimation of Toxocara canis prevalence in dogs, environmental egg contamination and risk of human infection in the Marche region of Italy.

The human risk of infection with larvae of Toxocara canis was estimated in people from the Marche region of Italy. This region includes both urban and rural areas and its inhabitants frequently keep dogs for company, hunting, as guardians or shepherds. T. canis infection was diagnosed in 33.6% out of 295 dogs examined. Nearly half of the dogs (48.4%) living in rural areas were found T. canis positive, compared to about one-quarter of the dogs (26.2%) from urban areas. Analysis by provenance and role revealed the highest infection rate in rural hunting dogs (64.7%) and the lowest in urban companion dogs (22.1%). According to questionnaire data, the peridomestic environment, i.e. gardens and dog pens, is the most important defecation site in both rural and urban areas. Since over 40% of the dogs who defecate in dog pens are infected and 24% of urban and 47% of rural dogs who leave their droppings in the house surroundings harbour the parasite, it is clear that these environments may constitute sites of zoonotic risk. Our analysis of soil samples from 60 farms confirmed the high contamination level, revealing positive soil samples in more than half of the farms. Substantial egg contamination was also found in urban areas, as 3/6 parks examined were Toxocara spp. positive. Finally, our serological findings indicate that human infection actually occurs in the area: 7 out of 428 adults examined (1.6%) had very high levels of antibodies to T. canis antigen, suggesting a previous contact with the Larva migrans of the nematode.

Animals↗

Transmission trials, ITS2-PCR and RAPD-PCR show identity of Toxocara canis isolates from red fox and dog.

Toxocara canis isolates from dog and from red fox were compared in transmission trials and with molecular analysis using RAPD-PCR technique and comparison of the ITS2 sequence. After oral infection of bitches with 20,000 embryonated T. canis eggs of vulpine and canine origin, the vertical transmission to pup's was examined. All animals of both groups developed typical clinical symptoms of toxocarosis. The haematological, serological, parasitological and post mortem results showed no differences between both isolates except for the infectivity of T. canis stages in mice where the fox isolate showed a significant higher infectivity than the dog isolate. The RAPD-PCR showed a similarity coefficient of 0.95, similar to the range of intraspecific variation in Toxocara cati and Toxascaris leonina specimens as outgroups. The ITS2 comparison showed a 100% identity between both isolates with no intraspecific variations. Therefore, the study shows that the fox and the dog isolate of T. canis were identical in infectivity, transmission and molecular structure; a host adaptation could not be found and the fox has to be seen as a reservoir for T. canis infections in dogs. Considering the increasing number of foxes in urban areas the importance of helminth control in dogs is stressed.

Animals↗

Toxocara cati: an underestimated zoonotic agent.

The role of Toxocara cati as a zoonosis is reviewed. It is suggested that, despite case histories of human infection in the literature, historical factors have led to T. cati being under-recognized as a zoonosis, particularly when compared with the prominence given to Toxocara canis in dogs. Differentiation of the two infections remains challenging even today. It is recommended that further work be conducted to facilitate differentiation so that the importance of T. cati as a zoonosis can be clearly defined.

Animals↗

A new technique for the recovery of Toxocara eggs from soil.

A new flotation technique for the extraction of Toxocara eggs from soil has been devised. It has a fairly high recovery efficiency for Toxocara eggs, can be used for replicated quantitative experiments, and may also be used for recovery of other ascarid eggs from soil.

Animals↗

Toxocara infection in pigs. The use of indirect fluorescent antibody tests and an in vitro larval precipitate test for detecting specific antibodies.

An in vitro larval precipitate test using second-stage Toxocara canis larvae and an indirect fluorescent antibody (IFA) test employing cuticles of T. canis larvae as antigen were evaluated using antisera produced in pigs experimentally infected with T. canis, T. cati, Ascaris suum, Toxascaris leonina and Parascaris equorum. The former test was both specific and sensitive and is suggested as a reliable and simple method of detecting Toxocara antibodies in pigs. The latter test was considered unsuitable because of cross-reactions that occurred when sera from pigs infected with other ascarids were tested. An IFA test for Ascaris antibodies, employing cuticles of A. suum larvae as antigen, is described. The degree of specificity of this test suggests that it may be of value in the detection of antibodies to Ascaris in pigs under natural conditions.

Animals↗

A seroepidemiological study of human Toxocara infection in the Slovak Republic.

Sera of 908 blood donors, average age 35 years, living in towns and villages of the Slovak Republic were tested for Toxocara canis antibodies using an ELISA-IgG test. The seroprevalence in the healthy population was estimated at 13.65%. Antibodies were predominantly demonstrated in the sera from persons living in rural communities (17.09%) compared to the sera investigated from urban communities (11.82%). Women had a significantly higher seropositivity than men. The serological response in 2703 sera from patients suspected of toxocarosis was compared with their clinical manifestation, sex and age. Toxocara antibodies were found in 27.41% of suspected patients. Although the seroprevalence in adults was significantly higher among women than men, in children, boys had a higher seroprevalence than girls. The main clinical manifestations were leucocytosis and eosinophilia (46%), ocular disorders (36%), lung, liver and neurological disorders (26%). The most severe forms of this disease were observed in 2 to 5-year-old children with geophagy.

Adolescent↗

Serological response of ascarid-free dogs to Toxocara canis infection.

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.

Animals↗

Population dynamics of Toxocara canis in pigs receiving a single or multiple infection.

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.

Animals↗

Molecular characterization of a Toxocara variant from cats in Kuala Lumpur, Malaysia.

The ascaridoid nematode of cats from Kuala Lumpur, Malaysia, previously identified morphologically as Toxocara canis, was characterized using a molecular approach. The nuclear ribosomal DNA (rDNA) region spanning the first internal transcribed spacer (ITS-1), the 5.8S gene and the second internal transcribed spacer (ITS-2) was amplified and sequenced. The sequences for the parasite from Malaysian cats were compared with those for T. canis and T. cati. The sequence data showed that this taxon was genetically more similar to T. cati than to T. canis in the ITS-1, 5.8S and ITS-2. Differences in the ITS-1 and ITS-2 sequences between the taxa (9.4-26.1%) were markedly higher than variation between samples within T. canis and T. cati (0-2.9%). The sequence data demonstrate that the parasite from Malaysian cats is neither T. canis nor T. cati and indicate that it is a distinct species. Based on these data, PCR-linked restriction fragment length polymorphism (RFLP) and single-strand conformation polymorphism (SSCP) methods were employed for the unequivocal differentiation of the Toxocara variant from T. canis and T. cati. These methods should provide valuable tools for studying the life-cycle, transmission pattern(s) and zoonotic potential of this parasite.

Animals↗

Allogeneic bone marrow transplantation-mediated transfer of specific immunity against Toxocara canis associated with excessive IgE.

A girl with myelodysplastic syndrome (RAEB-T) received HLA-identical bone marrow from her younger brother after myeloablative treatment with busulfan and cyclophosphamide. After bone marrow transplantation, fever, exanthema, pruritus, and a pulmonary infiltrate were treated symptomatically. Bacterial cultures remained negative. Leukocyte engraftment began on day 10, and all blood cell populations proved to be of donor origin on FISH analysis. Increasing IgE levels (21 000 U/ml) on day 14 after BMT, positive RAST, specific IgG-antibodies, and missing Toxocara (T.) canis antigens in the recipient indicated donor-derived seroconversion. Before BMT, the recipient had been negative for T. canis in routine parasitological screening, and the donor proved to be positive for T. canis antibody by ELISA. This report suggests that the transfer of IgE immunity in the absence of detectable antigens may be responsible for IgE-mediated symptoms consistent with toxocara infection and confirms the need for parasite screening in donor medical examinations.

Adoptive Transfer↗

Identification, characterization and expression of Toxocara canis nematode polyprotein allergen TBA-1.

We have cloned the cDNA of TBA-1, the Nematode polyprotein allergen (NPA) of Toxocara canis and found it to be most similar to ABA-1, the Ascaris NPA, on the basis of amino acid sequence. We could study the antigenic properties of an E-coli synthesized fusion protein prepared with the cloned gene since no glycosylation site was expected from the deduced amino acid sequence. Although no IgE responses to TBA-1 were detected, recombinant TBA-1 was differently recognized by serum IgG antibodies when the recombinant TBA-1 was directly adsorbed vs when immobilized via a streptavidin linkage on polystyrene microtitre wells. One group of sera recognized TBA-1 directly immobilized while the second only recognized TBA-1 immobilized via streptavidin linkage. The former were from rodents immunized with a Toxocara sp. adult worm extract while the latter were obtained from rodents infected with T. canis larva or immunized with a Anisakis simplex L3 larval extract. These observations suggest that the two in vivo forms of TBA-1 are expressed, but during different stages of the parasite's life cycle.

Allergens↗

Toxocara canis: egg presence in Melbourne parks and disease incidence in Victoria.

PURPOSE: Toxocara canis can cause blinding eye disease. This study assessed the presence of T. canis eggs in soil from parks in Melbourne and also the incidence of presumed ocular toxocariasis in Victoria. METHODS: One hundred and eighty soil samples were collected from nine suburban locations in Melbourne, Australia. These were analyzed for the presence of T. canis eggs. A search of laboratory records of T. canis serology requests from Victorian patients over an 8-year period was performed. RESULTS: Only one soil sample was positive for T. canis eggs. Positive T. canis serology was reported in 13 samples from patients. These patients all had ocular features suggestive of T. canis infection. CONCLUSION: Toxocara canis eggs are rare in public parks in Melbourne and symptomatic ocular toxocariasis is uncommon in the Victorian population. The acquisition of the disease is unlikely to be from public parks.

Adolescent↗

Eosinophilic pleocytosis and myelitis related to Toxocara canis infection.

Toxocara canis causes the visceral larva migrans syndrome in which central nervous involvement is rare. We report the case of a 40-year-old woman presenting with a subacute weakness of the right leg and dysaesthesiae in the right Th8-Th10 dermatomas. Spinal magnetic resonance imaging examination showed abnormal hyperintensity within the spinal cord. Cerebrospinal fluid analysis revealed eosinophilic pleocytosis. Antibody titres to Toxocara canis were higher in the cerebrospinal fluid than in the serum. Treatment using mebendazole led to a complete clinical recovery, normalization of cerebrospinal fluid parameters and improvement in spinal magnetic resonance imaging abnormalities.

Adult↗

Toxocara canis infections in a pig model: immunological, haematological and blood biochemistry responses.

The immunological, haematological and enzymatic responses to the inoculation in pigs of 100,000 embryonated eggs of Toxocara canis were studied. Fifteen females were inoculated and three remained as controls. Haematological values were analysed from day 7 p.i. until day 126 p.i. In the inoculated group, white blood cells were raised on day 14 p.i. and eosinophil values on days 7, 14, 21, 35 and 49 p.i. showing significant differences compared with controls (P < 0.05). Absolute eosinophil counts (per ml) presented two rises, the first on days 7, 14 and 21 p.i. and the second on days 35 and 49 p.i. Blood biochemistry was maintained within normal values. Serological examination by ELISA to determine antibody levels against Toxocara canis L2/L3 excretory-secretory (ES) antigens showed values higher than the positive cut-off (1:32) from day 7 p.i. and until the end of the study on day 126 p.i., presenting two peaks: one on day 28 p.i. and the second covering days 49 to 56 p.i. Western blots of sera of inoculated animals presented, from day 7 p.i., two polypeptide bands of 55 and 70 kDa MW and, from day 56 p.i., an additional band of 120 kDa MW, all of which persisted until the end of the study. Immunological responses were sustained over time. No direct correlation was observed between the rise in eosinophils and antibody titres. To validate the conclusions, more studies are required on the polypeptide bands.

Animals↗

Toxocara eggs in the soil of public places and schools in Basrah, Iraq.

One hundred and eighty soil samples were collected from public places and school yards in urban and rural areas in Basrah between October 1991 and March 1992. Each sample was examined by zinc sulphate flotation and sedimentation. Toxocara eggs were found in 22 samples (12.2%). Of the recovered positive samples 10 contained viable and fully embryonated eggs. Viable eggs were observed in higher proportions of soils collected from the surroundings of Basrah slaughter-house, where there are large numbers of dogs and cats. The observed mean number of eggs per 5 g of soil was 2.8 +/- 1.9 S.D. The contamination rate in urban areas (17.3%) was significantly higher (P < 0.05) than that in rural areas (6.1%). Thirteen of 116 samples (11.2%) collected from public places and nine of 64 samples (14.1%) from school yards contained Toxocara eggs. Urban public places and schools have higher contamination rates (18.0% and 16.2%, respectively) than those in rural areas (3.6% and 11.1%), but the differences are significant (P < 0.05) between public places only.

Animals↗

A survey of Toxocara infections in cat breeding colonies in The Netherlands.

Faecal samples from 225 adults cats and 112 kittens and dust and soil samples from 25 catteries in the Netherlands were examined for Toxocara eggs. The results of this survey showed a low nematode infection rate in the investigated Dutch catteries since only four adult cats (2%) from two catteries (8%) were found to shed Toxocara cati eggs. No other helminth eggs were seen in the faecal samples. Nematode eggs were not present in the environmental dust and soil samples from houses and kennels; only Dipylidium caninum eggs were found in only two samples of household dust.

Animal Husbandry↗

Nephrotic syndrome associated with Toxocara canis infection.

This case report describes nephrotic syndrome in a 7-year-old boy coincident with Toxocara canis infection. This rare association was confirmed by elevated Toxocara-specific IgM titres. Treatment with corticosteroids resulted in remission of renal symptoms as well as abatement of the T. canis infection. The relationship between T. canis infection and glomerular disease is still unclear; nephrotic syndrome may be another manifestation of T. canis infection.

Animals↗