Experimental infection of Japanese quail with Toxocara canis larvae through earthworms

The migration and distribution of Toxocara canis larvae in the tissues of earthworms exposed to 5 x 10(5) infective eggs in soil, as well as the reinfectivity of the larvae in Japanese quail fed with three earthworms, were studied. The average number of larvae recovered from an earthworm gradually declined from 9 +/- 3.21 to 4.5 +/- 2.00 at 4-16 days. No larvae were recovered at Day 20. When compared between three portions of the earthworms' body, the middle part showed the greatest recovery in comparison with the anterior and posterior parts. At necropsy, the average larval burden of quail at 15 days post-infection was 6 +/- 3.60 in the liver and no larvae were recovered from other tissues. The role of the earthworm in relation to paratenism is discussed.     

Acquired immunity to Toxocara canis infection in mice

Acquired immunity develops against Toxocara canis infection in mice, and NIH mice are more immunoresponsive than CD1 mice. Twice infected NIH female mice showed 27% reduction in the total larval recoveries compared with non-sensitized controls. Twice-infected NIH male, and CD1 (both sexes) mice showed a negligible reduction in the total recoveries, though a significant (P less than 0.05) number of larvae were retained in the liver compared with the non-sensitized controls. All twice-infected mice showed a significant reduction in the number of larvae recovered from the brain compared with once-infected mice. Vaccination using ultraviolet irradiated embryonated eggs gave the best protection against reinfection. Excretory/secretory antigen afforded less protection, whilst whole adult worm vaccine and whole L2 culture vaccine gave no protection. Vaccinated mice had a higher 'free:penetrating ratio' of larvae in their intestine than similarly challenged but non-vaccinated mice. When the ileum was examined histologically 9 h post-infection, an inflammatory reaction was seen around the penetrating larvae in the sensitized and vaccinated mice but not in untreated controls, suggesting a role played by the intestine in the resistance against T. canis infection in mice.     

Experimental Toxocara canis infection in cynomolgus macaques (Macaca fascicularis)

Visceral larva migrans was produced in 16 cynomolgus macaques (Macaca fascicularis) experimentally inoculated with 45,000 embryonated eggs of Toxocara canis as a single bolus or in 3 divided doses. The hematologic and serologic changes were similar to those observed in children with visceral Toxocara infection. Neurologic signs developed in 3 animals and were characterized by ataxia and nystagmus. Growth rates were diminished in inoculated animals when compared with the rates in noninoculated controls. Diagnostic antibody titers as measured by enzyme-linked immunosorbent assay were noted in all inoculated macaques by the 4th week, and these titers persisted during the 7-month period of observation. Antibody to Toxocara was not detected in the aqueous or vitreous humors. Lesions comprised severe granulomatous hepatitis and encephalomyelitis. Intraocular lesions associated with larval migration were not observed.     

Effects of milbemycin on adult Toxocara canis in dogs with experimentally induced infection

To determine the efficacy of a formulation of milbemycins in treating patent infection with Toxocara canis, 8 male and 7 female, 10-week-old, ascarid-free Beagles each were given 125 embryonated eggs of T canis. All dogs developed patent infection within 56 days. On post-infection day 70, the dogs were assigned to 1 of 3 groups of 5 dogs each; members of 1 group were given a placebo, while dogs of the other 2 groups were given either 5.68 or 34.08 mg of the milbemycin formulation, respectively. In both groups of dogs given the drug, the number of eggs passed per gram of feces decreased precipitously. However, a few eggs still were found in the feces of several dogs of each group on the day of necropsy (postinfection day 75). Worms or fragments of worms were passed by the treated dogs from the day of treatment until the day on which necropsy was performed; however, most worms were passed during the first 2 days after treatment. At necropsy, only dogs of the control group were found to harbor adult T canis.     

Larval development of Toxocara canis in dogs

The parasitic roundworm Toxocara canis is present in dog populations all over the world. Due to its zoonotic potential, this roundworm is of special interest not only for veterinarians, but also for medical practitioners. In the present review, current knowledge of infection routes and the subsequent development of larvae within the canine host is summarised. Furthermore, information about the clinical, pathological, enzymatic, haematological and histopathological changes was collected, giving a broad overview of current knowledge of the infection. Although the data collected over the years give an idea of what happens during the larval development of T. canis, many questions remain open. Nevertheless, it is important that we continue our efforts to further understand the biology of this versatile and compelling parasite and try to improve and optimise strategies to prevent the infection in dogs and thereby to protect humans from this infection.     

Periparturient immunosuppression in the bitch and its influence on infection with Toxocara canis

Pregnancy and lactation in dogs induced a marked suppression of their immunological responsiveness as judged by in vitro phytomitogen- and Toxocara canis antigen-induced lymphocyte transformation. In addition, the eosinophilia otherwise seen following infection with T. canis was suppressed during the periparturient period. Coincidental with this periparturient immunosuppression was the occurrence of heavy infections with T. canis in the intestine of lactating bitches and the establishment of such infections perhaps may be related to the suppressed immune reactivity observed. These infections in the bitch appeared to be acquired primarily through the ingestion of immature stages of T. canis passed in the faeces of their puppies. The results demonstrate that the lactating bitch can be a major source of contamination of the environment with eggs of T. canis.     

Variations in innate resistance to experimental Toxocara canis infection in two strains of mice

Strain-, sex- and age-based resistance of mice to Toxocara canis infection was demonstrated. Infection doses of similar numbers resulted in significantly (P less than 0.05) greater worm establishment in CD1 mice than in NIH mice, as well as a greater heterogeneity of responses between individuals. Male mice had significantly (P less than 0.05) fewer larvae in the brain and more larvae in the liver than female mice, when mice of greater than or equal to 3 months old were used. Appreciably more larvae were retained in the liver of male CD1 mice than in NIH mice. Orchidectomized mice had higher levels of brain infestation and a lower larval recovery from the liver compared with intact male mice. Infections in young mice of both strains showed more diversity in establishment and migration pattern between individuals than in older mice. Despite these differences in distribution of larvae and in the numbers recovered from experimental infections, the acute and chronic clinical signs observed in the two strains were very similar.     

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.     

Seroprevalence of Toxocara canis in sheep in Wales

Antibody levels to Toxocara canis L2 excretory/secretory antigens were examined by ELISA in 400 serum samples from sheep in Powys and Gwent, Wales. A positive OD value was set at the mean +/-3S.D. of 45 control samples. Seroprevalence increased with age. Seven percent and 13% of 6-month-old sheep showed positive OD values as did 16% of 10-month-old, 27% and 31% of 15-month-old and 47% of cull ewes. Analysis of variance showed a significant increase in ELISA OD values among the seropositive sheep with increasing age of sheep.     

Expert opinion based modelling of the risk of human infection with H5N1 through the consumption of poultry meat in Germany

According to current scientific opinion the risk of human infection with H5N1 via preparation and consumption of poultry meat is negligible.This opinion has not yet been challenged by a formal risk assessment, due to the lack of empirical data. We have developed a scenario pathway model as a conceptual framework for a formal assessment of the H5N1 risk to humans through consumption of poultry meat and parameterise the model using information derived from expert opinions. The aim of this study was to investigate whether the notion of an overall negligible risk via the oral infection route is consistent with ad hoc data and expert opinions on the relevant parameters of the model.The model is mainly based on expert opinion. A stochastic Monte-Carlo simulation was conducted which took into consideration (amongst others) the exposure and infection of chicken (broiler and layer), turkeys, ducks and geese, the probabilities of detection prior to slaughter, virus survival and contamination during slaughter, as well as during the cutting and preparation of meat in commercial plants and in private households, respectively. The empirical consumption pattern for poultry meat in Germany was taken into account in the simulation.The results show that the risk for the individual consumer is practically zero whereas up to 23 cases per year in Germany might occur if the upper (more pessimistic) ranges of the expert opinions apply.The finding of a low but non-negligible risk to the population is discussed in relation to the epidemiological information available from recent outbreaks in South East Asia.     

Contamination of dog hair with eggs of Toxocara canis

Toxocara canis, the common intestinal nematode of dogs and foxes, is the parasite responsible for human toxocarosis. It has recently been shown that dogs may harbour eggs of the parasite in their fur. To further investigate this claim a population of 100 stray dogs was examined to establish the prevalence and intensity of adult toxocaral worm infection in the intestines and eggs harboured in the hair. A novel method of washing the eggs from the hair was used. Sixty-seven percent of dogs were found to have T. canis eggs on their hair with a mean egg retrieval of nearly 584 eggs per gram from positive dogs. The age of the dog was found to be the only significant factor to influence the prevalence and intensity of eggs, with 95% of all the eggs recovered found on puppies. Thirty-nine percent of dogs were found to have adult T. canis worms in their intestine, although a significantly higher percentage of puppies (80%) were infected with worms than adults (22.5%). Puppies also had more worms per infection than adults and have a strong positive correlation between egg and worms numbers whereas adults did not. These studies show that stray dogs, particularly puppies, potentially harbour considerable numbers of eggs on their hair, at densities far higher than those reported in the soil or the general environment.     

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.     

Experimental Ascuris-induced airway hypersensitivity in sheep: no serological evidence of association with Toxocara canis infection

Abstract

Extract

The finding that 70% of sheep from the Manawatu district were hypersensitive to challenge by inhalation of Ascaris suum antigen, regardless of their previous history of exposure to pigs(1 Chen, W, Pack, RJ, Alley, MR, Carr, DH and Manktelow, BW. 1990. Airway hypersensitivity induced by Ascaris scum extract in New Zealand Romney sheep. New Zealand Veterinary Journal, 38: 5741–5741.  [Google Scholar]), has raised the possibility that these sheep had been sensitised by exposure to the common dog nematode Toxocara canis.     

Toxocara canis infection induces antigen-specific IL-10 and IFNgamma production in pregnant dogs and their puppies

Toxocara canis (T. canis) is originally a parasite of canine bitches and their pups. The pathogenicity of T. canis infection is enhanced during pregnancy and puppyhood. The aim of this study was to investigate if modification of IFNgamma and IL-10 secretion occurs during infection in pregnant dogs and puppies. Analysis of cytokines secreted could let us hypothesize a role for IL-10 and/or IFNgamma in T. canis infection. We tested T. canis-specific production of IFNgamma and IL-10 by lymphocytes of pregnant dogs and their puppies after in vitro re-exposure to purified excretory/secretory antigen (ESAg) from T. canis. Blood mononuclear cells (BMC) isolated from pregnant dogs and their puppies were cultured in the presence of ESAg. Cultures' supernatants were tested for cytokine levels by ELISA. Results obtained showed that IL-10 concentrations increased during pregnancy in infected animals and in the meantime IFNgamma production decreased. In puppyhood, we observed that, IL-10 concentration decreased with the age of puppies mainly in infected animals while IFNgamma increased. In conclusion, our data suggests that BMC of infected dogs have a particular modification of IL-10 and IFNgamma synthesis. These data could be the basis to design immunotherapeutic approaches.     

犬弓首蛔虫TcCPG1的表达及其与几丁质的结合特性

为探讨犬弓首蛔虫（Toxocara canis,T.canis）软骨素蛋白多糖TcCPG1的表达特征及其与几丁质的结合方式,为犬弓首蛔虫软骨素蛋白多糖的功能研究提供依据,笔者克隆T.canis软骨素蛋白多糖1基因（Tc-cpg-1）并原核表达重组蛋白TcCPG1,采用镍柱亲和层析技术对重组蛋白进行纯化,并与几丁质进行结合试验;采用实时荧光定量PCR（quantitative real-time polymerase chain reaction,qRT-PCR）进行Tc-cpg-1基因的转录水平的性别和组织差异表达分析。结果显示：Tc-cpg-1基因编码区（coding sequence,CDS）的长度为1 251 bp,共编码417个氨基酸;SDS-PAGE电泳结果显示重组蛋白TcCPG1以包涵体形式存在,相对分子质量大小约为70 ku;对表达条件进行优化后,以0.8 mmol·L^-1 IPTG于16℃诱导14 h时可获得大量目的蛋白;利用镍柱亲和层析纯化可以获得具有较高纯度的目的蛋白,与几丁质进行结合试验,验证了重组蛋白TcCPG1能够与几丁质结合;通过qRT-PCR对Tc-cpg-1基因转录水平的组织差异性进行分析,发现Tc-cpg-1在雌虫中高量表达,尤其在雌虫的生殖组织中表达量最高,表明Tc-cpg-1可能参与了T.canis雌虫的生殖过程,推测TcCPG1可能通过与卵壳中几丁质的相互作用从而影响T.canis的胚胎和生殖发育过程。