Hair coat contamination with zoonotic helminth eggs of farm and pet dogs and foxes

Infections of dogs with Toxocara canis and Echinococcus multilocularis pose an infection-risk particularly for contact persons. We examined specimens of hair coat and faeces of 124 farm dogs, 118 household dogs, 49 kennel dogs, 15 puppies from two litters, and 46 red foxes. Microscopically identified eggs of Toxocara or taeniids were further investigated by species-specific PCRs. In farm dogs, eggs of E. multilocularis or T. canis were identified in each 2.4% of faecal samples, eggs of T. cati (gastrointestinal passage) in 7.3%, respectively. Household dogs excreted eggs of T. canis (0.8%) and of T. cati (2.5%). In kennel dogs, eggs of T. canis (4.1%), but not of T. cati were detectable. Coat samples contaminated with eggs of Toxocara spp. were found from farm dogs (5.6%), household dogs (1.7%) and kennel dogs (2.0%). Taeniid eggs were isolated from the coat samples from only two farm dogs (1.6%); a molecular species determination was not achieved. In six intrauterinely infected puppies, Toxocara-eggs were found in 17/38 samples taken within six weeks. No intact Toxocara eggs could be isolated from the coat of nine puppies from a second litter 13 days after deworming. Of the 46 red foxes investigated (dissection and faecal samples) 13 (28.3%) were infected with E. multilocularis and 20 (43.5%) with Toxocara. Eggs of taeniids and Toxocara were found in 13% (in three cases confirmed as E. multilocularis) and 21.7%, respectively, of the coat samples. None of the retrieved Toxocara eggs in the coat samples were embryonated. Thus, an infection of humans through the transmission of E. multilocularis eggs after direct contact with dogs or foxes is conceivable, whereas a corresponding infection risk by Toxocara eggs must be critically challenged.     

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.     

Toxocara eggs shed by dogs and cats and their molecular and morphometric species-specific identification: is the finding of T. cati eggs shed by dogs of epidemiological relevance?

Intestinal infections with Toxocara cati and Toxocara canis in their definitive host (felids and canids, respectively) are diagnosed by egg identification in faeces using coproscopical techniques. The Toxocara species is assumed to comply with the species from which the examined faeces were obtained, i.e. T. cati in cats and T. canis in dogs. We isolated and measured Toxocara eggs from faecal samples of 36 cats and 35 dogs from Switzerland and identified the Toxocara species by PCR. Amongst the isolates originating from dogs, 24 (68.5%) were determined as T. canis and 11 (31.5%) as T. cati. In all samples originating from cats, only T. cati was identified. Based on PCR identification, eggs of T. canis (n=241) and T. cati (n=442) were measured, revealing statistically significant different (p<0.001) mean sizes of 62.3 by 72.7 μm for T. cati and 74.8 by 86.0 μm for T. canis eggs. Considering that coprophagy is not unusual for dogs, a considerable percentage of Toxocara infections coproscopically diagnosed in dogs, as well as assumptions on anthelminthic resistance in regularly treated dogs, might in fact relate to intestinal passages of eggs following the uptake of other animals' faeces.     

Differentiation of Toxocara canis and T. cati eggs by light and scanning electron microscopy

In this study, we investigated the morphological identification of Toxocara canis and T. cati eggs on the basis of light and scanning electron microscopic (SEM) observations. T. canis and T. cati eggs used in this study were recovered from the uteri of respective gravid female worms. Measurement of egg size was not helpful in the differentiation of these species, because approximately 90% of eggs measured were of similar size. Using SEM, we were able to differentiate T. canis eggs from T. cati eggs based on their respective characteristic surface structures. Both species have subspherical eggs with markedly pitted surfaces like those of a golf ball, but the surface pitting in T. canis is more coarse than that in T. cati. In this study, however, these differences were not absolute, as 16% of T. canis and 29% of T. cati eggs showed surface pitting that was uncharacteristic of their species. Of the 16% of T. canis eggs that could not be differentiated by surface structure, 3% had pitting resembling T. cati, and the remaining 13% showed intermediate type surface pitting. Similarly, 5% of T. cati eggs resembled T. canis, and 25% of these were of intermediate type. Light microscopic observation yielded results similar to those of SEM, indicating that light microscopy is also a useful tool for the identification of Toxocara eggs.     

Distribution of Toxocara infection in the environment and in definitive and paratenic hosts in Estonia

The aim of the study was to elucidate the distribution and possible transmission routes of Toxocara spp. infection in Estonia. Out of 454 faecal and sand samples collected from park lawns and sandpits in the town of Tartu, 19 were Toxocara positive (4.2%). Out of the 45 sandpit samples 17.8% were Toxocara positive. Cat faeces was found in 21 sandpit samples. Parasitological necropsies were performed on 41 euthanised stray dogs and 27 cats in the Tallinn Dog Home. Additionally, 13 wild free-roaming brown rats (Rattus norvegicus) were captured from the Tallinn Dog Home territory, necropsied and studied for the presence of Toxocara larvae. Toxocara canis adults were found in 14.6% of the dogs and Toxocara cati (syn. mystax) adults in the small intestines of 48.2% of the cats examined. Larval infection was detected in the kidney and liver in 5 dogs (12.2%). Our study demonstrated only low-level larval Toxocara infections in adult dogs. Toxocara larvae were not found in cats and brown rats. According to the results of this study, cats more often carry Toxocara infection than dogs. Under our conditions, stray and free-roaming cats are the main contaminators of the environment with Toxocara eggs. Children playing in sandpits are the main risk group for larval toxocarosis.     

Tolerance to low temperatures of Toxocara cati larvae in chicken muscle tissue

Infectivity of Toxocara cati larvae in muscle tissue of chickens after storage at 4°C and -25°C was assessed in a mouse bioassay to provide information on the risk of meat-borne toxocarosis. Muscle tissue samples of 30-day old T. cati infections were stored at 4°C for 14 and 28 days and at -25°C for 12, 24 and 48h, whereafter, larvae were released by digestion. For each experimental group, the released larvae were inoculated in six mice. After 15 days, mice were euthanized and larval burden was assessed by digestion. In the control group (no storage of the infected chicken meat), 47.9% of the inoculated larvae established in mice, whereas storage of meat at 4°C for 14 days or 28 days reduced the recovery to 24.1% or 3.3%, respectively. Muscle larvae exposed to -25°C for 12, 24 or 48h did not establish in the mice. The observation that larvae retain infective after refrigeration at exposure in 4°C for 28 days, emphasize the zoonotic potential of poultry meat as a causative agent of human toxocarosis.     

Circulation of Toxocara spp. in suburban and rural ecosystems in the Slovak Republic

Toxocara spp., the common roundworms of domestic and wild animals, are the causative agents of larval toxocarosis in humans. The migration of Toxocara larvae in men causes clinical syndrome, called larva migrans visceralis or larva migrans ocularis. The objective of the present work was to investigate the prevalence of toxocarosis in dogs, red foxes (Vulpes vulpes), and small mammals in the Slovak Republic. T. canis infection was diagnosed in 16.6% out of 145 dogs examined. Young animals showed significantly higher positivity (50.0%) than adults (12.4%). Coprological investigation of 310 red foxes showed 8.1% prevalence of T. canis, with higher occurrence in animals from rural conditions (8.6%) than from suburban environment (5.6%). Out of 710 small mammals examined, anti-Toxocara antibodies were detected in 7.7% of the animals. The most frequently seropositive species was Apodemus agrarius (15.9%). Seropositivity of small mammals from suburban localities was higher (10%) compared with rural areas (5.8%), with adult animals exhibiting higher seropositivity (8.0%) than subadults (6.8%). Our results have confirmed the importance of dogs, red foxes and small mammals in circulation of this serious helminthozoonosis in suburban and rural ecosystems.     

Changes to plasminogen activators and matrix metalloproteinase-9 in dogs with toxocarosis

The relationship between proteolytic enzymes and hematological response to infection was studied in five 1-month-old dogs inoculated experimentally with 2000 eggs of Toxocara canis. Moderate leukocytosis and marked eosinophilia occurred 14 days post-infection with T. canis. Plasminogen activators (PAs) and matrix metalloproteinase-9 (MMP-9) activity in serum was significantly different in dogs infected with T. canis, compared with controls. Urokinase-type PA (uPA) activity was positively correlated with eosinophilia, and tissue-type PA (tPA) and MMP-9 activity was negatively correlated with eosinophilia. However, there was no correlation between inflammation and MMP-2. The use of uPA, tPA or MMP-9 proteolytic enzymes as laboratory reference markers for toxocarosis requires further study.     

Ascarid nematodes in domestic and wild terrestrial mammals

The biology of the ascarid nematodes has been discussed in the context of their important economic role in farm animals, pet animals and zoo animals with special attention to carnivores and primates. In farm animals, infection with the most common roundworm of horses (Parascaris equorum) and swine (Ascaris suum) depend on many factors such as environmental conditions (larval development in the egg and egg survival), age of the host, breed, husbandry system, hygiene and treatment schedule. The monoxenic ascarids Toxocara canis and T. cati are the most important nematodes in carnivorous animals (dogs, cats, foxes) and carnivores in the zoo. In the period of March 2000 till March 2001, 57.1% of examined representatives of Felidae, Ursidae and Canidae in the Zoological Garden of Wroc?aw were found to be infected with ascarids (T. canis, T. cati, Toxascaris leonina). The prevalence of T. canis in the Canidae was 66.7%, of T. cati in the Felidae was 14.3%, while 57.1% of the Felidae carried T. leonina infection. Ascaris lumbricoides, a typical parasite of primates, was found in some gorillas, chimpanzees and orang-utans during parasitological survey.     

Larval toxocarosis in sheep: the immunohistochemical characterization of lesions in some affected organs

Morphological and immunohistochemical responses of lambs following oral infection with 10,000 infective Toxocara canis (T. canis) eggs were studied up to 28 days post-infection (pi). The small intestine, liver, lungs and brain of both infected and control lambs were examined using the routine histological methods for paraffin sections and immunohistochemical techniques for frozen tissue sections. Eosinophil-rich hepatic granulomas and diffuse T. canis-induced pulmonary inflammation were the most prominent pathological features. CD2+, CD4+, CD8+, IgM bearing cells, and macrophages were demonstrated in the liver granulomas. The observed histomorphologic changes were similar to other paratenic hosts. It was concluded that larval toxocarosis followed the classical migratory path in the infected lambs.     

Helminth and protozoan parasites in dogs and cats in Belgium

This study investigates the level of helminthic and protozoal infestation over the last 10 years in strays, well-cared-for dogs and cats. Determination of the prevalence of infections was based either on faecal examination or on worm counts at necropsy. Of 2324 faecal flotations (NaCl sp.gr. 1.20) of stray dogs, 34.2% had eggs or proglottids of one or more worm species consisting of Toxocara canis (17.4%), Toxascaris leonina (10.1%), Uncinaria stenocephala (11.4%), Trichuris vulpis (7.0%) and cestodes (2.1%). Isospora oocysts were observed in 5.2% of the dogs. The data on the distribution of the various worm species in the positive dogs indicate that T. canis eggs were by far the most common (50.9%). Necropsy data from 212 infected dogs indicate that 38.9% were infected with T. canis and 33.7% with T. leonina. The overall prevalence of worm infestation of 246 well-cared-for kennel dogs, based on worm egg counts by the McMaster technique, was 36.1%. Of 30 feline faecal samples examined by flotation, 83.3% were positive for parasites, including Toxocara cati (60%), Ancylostoma tubaeformae (36.6%), Taenia (Hydatigera) taeniaeformis (20%) and coccidia (30%). Toxocara cati was the most frequently found worm species at the necropsy of 25 cats (52%). Toxoplasma was not observed.     

Ascarid infections of cats and dogs

The ascarids Toxocara canis, Toxocara cati, and Toxascaris leonina are probably the most common gastrointestinal helminths encountered in small animal practice. Both T. canis and T. cati can cause serious disease in kittens and puppies; T. leonina is generally less pathogenic. Prenatal transmission assures that virtually all puppies are born infected with T. canis. Transmammary transmission is probably the major route of infection for kittens with T. cati. In addition, all three species of worm produce resistant eggs and use paratenic hosts to facilitate transmission. Much is now known about the complex biology and life history of T. canis. However, many questions, such as those concerning the mechanisms of larval survival within host tissues and of larval reactivation and migration during pregnancy, await further study. The mechanism of resistance to ascarid infections in cats and dogs has not been clearly defined. Ascariasis is traditionally thought to be a disease of young animals, with older animals being considered immune. However, at least in the case of T. canis, adult dogs can be repeatedly infected. A wide range of anthelmintics is available with extremely high efficacy against patent ascarid infections. The problem of prenatal infection with T. canis may be overcome by strategic use of the newer benzimidazole-carbamates, and the production of ascarid-free puppies now seems possible. However, complete larvicidal activity against somatic stages has not been convincingly demonstrated. Visceral larva migrans-like syndromes are now being recognized in dogs and cats. In addition, visceral larva migrans in children due to T. canis continues to be a significant zoonotic disease in North America and underscores the need for the veterinary profession to control ascarid infections in cats and dogs at every opportunity.     

Prevalence of Toxocara spp. and other parasites in dogs and cats in Halifax, Nova Scotia.

Several cases of visceral larva migrans syndrome in children led to an investigation of the ova of Toxocara spp. and other species in stool specimens from stray dogs and cats in Halifax, Nova Scotia. Fecal samples from 474 stray dogs and 299 stray cats were examined. Ova of Toxocara canis or cati were present in stools from 26.6% of the dogs and 25.1% of the cats. In dogs the prevalence of Toxocara canis was noticeably greater in puppies (56.1%) than in mature animals (11.9%).     

Prevalence of chlamydia, toxoplasma, toxocara and ringworm in farm cats in south-west England

The prevalence of infection with Chlamydia psittaci, Toxoplasma gondii, Toxocara cati and Microsporum canis was examined in 51 cats on 22 sheep farms in the Bristol area. Serum antibody to C psittaci and T gondii was present in 45 per cent and 47 per cent of cats, respectively. At the time of sampling C psittaci was isolated from 6 per cent of the cats, T cati was identified in 63 per cent of faecal samples but neither T gondii nor M canis was isolated. When examined according to the farm of origin, 22.7 per cent of farms had cat populations with no evidence of infection with C psittaci or T gondii. Of the remainder, 45.5 per cent supported cat populations with evidence of both infections and 31.8 per cent had evidence of T gondii infection alone. None of the farms had cat populations with evidence of C psittaci infection alone. Two of the cats infected with C psittaci were excreting viable organisms in the faeces. The possible significance of this to the epidemiology of ovine enzootic abortion is discussed.     

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.     

Anthelmintic efficacy of flubendazole paste against nematodes and cestodes in dogs and cats

Small dogs and cats, naturally infected with nematodes and cestodes, were used in a critical test under laboratory conditions to determine the palatability and efficacy of flubendazole as a past formulation. Subsequently, a control test in dogs was conducted under field conditions. A 4.4% past formulation was given at a dosage of 22 mg/kg of body weight once a day for 2 or 3 consecutive days. In a critical test in dogs, the efficacy against Toxocara canis was 97.4% after a 2-day administration and 100% after 3 days. Toxascaris leonina seemed to be the most susceptible worm species, since either 2 or 3 treatments were 100% active. The efficacy against Uncinaria stenocephala was 97.5% after 2 treatments; the same dose level for 3 days improved the efficacy to 100%. The efficacy was 100% for the removal of Trichuris vulpis after a daily dosage for 2 days and 96.7% after 3 days. One of 2 dogs infected with Taenia pisiformis was cleared of the infection after a 2-day treatment, and 3 of 4 dogs were cleared after a 3-day regimen. All cats were cleared of Toxocara cati after 2 or 3 days of treatment. One of 2 cats infected with Hydatigera taeniaeformis was cleared of the infection after a 2-day treatment; a 3-day treatment in 7 cats was 100% effective. The results in the laboratory test in dogs were confirmed under field conditions by a control test, based on the reduction of eggs per gram of feces count after treatment. The paste formulation was well accepted by all dogs and cats without any side effects.     

A simplified method for hatching and isolating Toxocara canis larvae to facilitate excretory-secretory antigen collection in vitro

Human toxocariasis causes several dangerous syndromes that can involve the viscera, vision and central nervous system. Diagnosing toxocariasis requires the identification of antibodies against Toxocara canis or Toxocara cati excretions and secretions (ES). To obtain ES it is necessary to collect a large number of larvae. However, since the earliest work describing the culture of Toxocara larvae, few advances in the method have been made. It has been suggested that carbon dioxide triggers molecular mechanisms that enable nematode hatching. A similar hypothesis has been made regarding Giardia excystation. To test the hypothesis we used the Giardia excystation HBSS method to hatch embryonated T. canis eggs. We found that the HBSS method was more effective than the original De Savigny method. Our results suggest that both parasites require stimulation in an acidic environment, and the abrupt change to a basic milieu in duodenum. This physiological adaptation is successful to exploit the intestinal habitat.     

Haematological and serological data from dogs raised worm-free and monospecifically infected with helminths

Pups free from helminth infection were obtained by treating the bitch prior to parturition, and the bitch and her pups after whelping, with oxfendazole and praziquantel. They were then housed in an area free from contamination with helminth eggs. Freedom from infection was confirmed by necropsy examinations and by serological tests using the Enzyme Linked Immunosorbent Assay (ELISA). Groups of pups were infected with Toxocara canis, Echinococcus granulosus, Taenia ovis, T. pisiformis and T. hydatigena and bled at 5-day intervals. Haematological examinations and measurement of serum proteins were performed on blood samples obtained throughout 30 days of infection. Levels of serum proteins were not significantly altered in any infection. Pups infected with T. canis showed a marked increase in numbers of circulating eosinophils during the phase of larval migration through the liver and lungs; they also showed marked antibody responses to T. canis larval ES antigen. Haematological values were not significantly altered in any other pups.     

Investigations into the prevention of prenatal and lactogenic Toxocara canis infections in puppies by application of moxidectin to the pregnant dog

Aim of the investigation was to examine whether two administrations of moxidectin to pregnant dogs could prevent pre-natal and lactogenic infections of puppies with reactivated Toxocara canis larvae. Four pregnant beagles, infected experimentally with 20 000 embryonated eggs of T. canis, were treated subcutaneously with 1 mg moxidectin per kg body weight on days 40 and 55 of pregnancy (5-13 days before parturition). One further dam and its puppies served as untreated control. Two applications of moxidectin completely prevented pre-natal and lactogenic infections in the puppies. Neither intestinal stages nor somatic larvae were found in the dams or their corresponding puppies. All puppies and dams of the treatment group remained coproscopically negative until 42 days after parturition. The administration of moxidectin did not show any side effects in the dams. None of the puppies of the treated dams showed any pathological abnormalities. In the untreated dam one adult and 26 somatic larvae of T. canis were detected at necropsy. All puppies of the untreated dam showed a patent T. canis infection from day 28 post-natum (p.n.); 296 pre-adult and adult stages of T. canis were spontaneously eliminated and 51 intestinal stages and five somatic larvae of T. canis were recovered at necropsy. In contrast to the puppies of the treated dams all negative control puppies showed blood eosinophilia after parturition and elevated liver enzyme levels.     

Toxocara cati larvae persist and retain high infectivity in muscles of experimentally infected chickens

The distribution of Toxocara cati larvae in the organs of chickens (n=31), experimentally inoculated with 3000 embryonated eggs, was examined 1, 2, 3, 7, 29, 86 and 175-176 days post-infection (dpi), and the infectivity of recovered larvae was evaluated by bioassay in mice. The duodenum, liver, lungs, heart, brain, pectoral muscles (white meat), and hindlimb muscles (red meat) of the chickens were HCl-pepsin digested for larval recovery. Larvae were recovered from all chickens [mean=220.4 ± 114.9 (SD)], and although no decrease of total larval recovery was observed over time, predilection sites changed: liver (92.6% of larval recovery) at 1 dpi; lungs (77.3%) and liver (20.9%) at 2 dpi; lungs (80.8%), muscles (9.4%), and liver (8.5%) at 3 dpi; muscles (52.0%) and lungs (45.6%) at 7 dpi, whereafter most larvae were recovered from muscles: 29 dpi (99.5%), 86 dpi (99.3%) and 175-176 dpi (99.6%). In the bioassay, 52.9% of larvae from 175 to 176 days old infections in chickens established in mice, which demonstrates that T. cati larvae retain infective in the muscles of chickens for half a year. These results highlight the zoonotic potential of poultry meat as a causative agent of human toxocarosis.