The concept of One Health emphasizes the interdependence of human, animal and environmental health and is of growing significance, in part owing to the problems related to emerging infectious diseases of wildlife origin. Wild mammalian predators are a potential risk factor for transmission of zoonotic pathogens to domesticated animals and humans. This is especially relevant in rural areas, where transmission of zoonotic pathogens can occur particularly efficiently when free-ranging dogs are present. The main aim of this study was to determine helminth infections among wild mammalian predators and evaluate the overlap between helminth faunas of wild mammals and dogs. Scat samples of predators were collected in coastal areas of Western Estonia and genetic methodology applied for the correct identification of predator species from their scat. Parasitic helminths of mammalian predators in the scat samples were analysed and compared with dog data from a previous study. High helminth prevalence (~90%) was found in dominant predator species in the area, namely the red fox (Vulpes vulpes) and golden jackal (Canis aureus). Moreover, the helminth fauna of both wild species, including potentially zoonotic helminths, overlapped largely with that of rural dogs in the same area. The results, together with the ones from earlier parasitological studies among humans in Estonia, emphasize the potential risk of pathogen transmission from wild mammalian predators to dogs and from dogs to humans, making parasitic diseases of wildlife a One Health concern.

Toxocariasis is a zoonotic disease caused mainly by Toxocara canis and Toxocara cati and diagnosis in dogs and cats is an important tool for its control. For this reason, a new coprological loop-mediated isothermal amplification (LAMP) assay was developed for the simultaneous detection of these species. The primer set was designed on a region of the mitochondrial cox-1 gene. Amplification conditions were evaluated using a temperature gradient (52°C to 68°C), different incubation times (15–120 min), and different concentrations of malachite green dye (0.004–0.4% w/v). The analytical sensitivity was evaluated with serial dilutions of genomic DNA from T. canis and T. cati adult worms, and with serial dilutions of DNA extracted from feces using a low-cost in-house method. The specificity was evaluated using genomic DNA from Canis lupus familiaris, Felis catus, Escherichia coli, Toxascaris leonina, Ancylostoma caninum, Echinococcus granulosus sensu stricto and Taenia hydatigena. The LAMP assay applied to environmental fecal samples from an endemic area showed an analytical sensitivity of 10–100 fg of genomic DNA and 10−5 serial dilutions of DNA extracted from feces using the low-cost in-house method; with a specificity of 100%. Additionally, the total development of the assay was carried out in a basic laboratory and per-reaction reagent cost decreased by ~80%. This new, low-cost tool can help identify the most common agents of toxocariasis in endemic areas in order to manage prevention strategies without having to rely on a laboratory with sophisticated equipment.

Neuroinvasive larvae of the common dog and cat roundworms, Toxocara canis and Toxocara cati, may cause severe neurological and neuropsychological disturbances in humans. Despite their pathogenic potential and high prevalence worldwide, little is known about their cell-specific influences and cerebral host–pathogen interactions in neurotoxocarosis. To address this discrepancy, a co-culture system of viable larvae with murine neuronal (CAD), oligodendrocytal (BO-1) and microglial (BV-2) cell lines has been established. Additionally, murine adult brain slices have been co-cultured with Toxocara larvae to consider complex organotypic cell–cell interplay. Cytotoxicity of larval presence was measured enzymatically and microscopically. Microscopic evaluation using trypan blue exclusion assay revealed to be less reliable and sensitive than the lactate dehydrogenase activity assay. Ultimately, even low numbers of both T. canis and T. cati larvae have impaired survival of differentiated CAD cells, which morphologically resemble primary neurons. In contrast, viability of oligodendrocytal and microglial cells as well as brain slices was not impaired by larval presence. Therefore, immune-mediated mechanisms or trauma by migrating larvae presumably induce the in vivo pathology rather than acute cytotoxic effects. Conclusively, the helminthic larvae co-culture system presented here is a valuable in vitro tool to study cell-specific effects of parasitic larvae and their products.

The importance of larval viability in relation to the success of recovery techniques and to the interpretation of histological and migration studies was tested using mice infected with Toxocara pteropodis, T. canis and T. cati. Little difference was found in total numbers of viable larvae recovered from tissues incubated for 18 hours in 0·85% saline, 2% trypsin or 1% pepsin, but the rates of larval egress did vary. The recovery of dead larvae by digestion techniques was much lower. Larvae migrated in tissues undergoing fixation, which influenced histological findings. Larvae of T. canis did not undergo significant peritoneal migration in acutely-infected mice, but did penetrate their intestinal walls in large numbers within 4 hours of death. The implications of these findings in experimental studies are discussed.

Contamination of 13 sandpits in public parks in Hyogo Prefecture, Japan, by Toxocara eggs and animal faeces was studied. All 13 sandpits were contaminated with faeces. The mean number of faeces found in 1 m2 of the sandpits was 35, from which the number of faeces in each sandpit was estimated to be 261 to 2205. Toxocara eggs were detected in 12 of the 13 sandpits, each examined 4–15 times; the overall prevalence was 92%. The eggs were not always distributed in the same pattern, but in general, eggs were numerous on the sandpit surface but also present at depths of about 35 cm. Of the eggs recovered, 63% were fully embryonated. Of the 13 sandpits examined, two were heavily contaminated by eggs, and all but one of the others were mildly contaminated. The difference between heavy and light contamination was not related to differences in the environment or in the number of faeces; the cause is unknown.

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.

The activities of the enzymes superoxide dismutase (E.C.:1.15.1.1) and catalase (E.C: 1.11.1.6) were studied in purified extracts of four nematodes: Ascaris suum, Toxascaris leonina, Toxocara canis and T. cati adult males and females. No catalase activity was found in any of the extracts. The results reveal that the SOD activities of the four parasites presented species differences and also sexual differences within each species. Polyacrylamide gel electrophoresis pattern analysis confirmed that the mobilities, widths and band intensities varied according to the species and sex of the parasite from which the enzyme was obtained.

Studies were conducted on the role of small mammals in maintaining toxocariasis foci in urban, rural and montane biotopes. The lowest relative density of small mammals was recorded in the urban locality and the highest in the rural and montane localities. Anti-Toxocara antibodies were most frequently detected in synanthropic and hemisynanthropic species Mus musculus, Apodemus agrarius and Micromys minutus – 32·0, 30·4 and 25·0%, respectively. The highest seropositivity was found in small mammals from the urban and rural localities – 22·2 and 21·6%, respectively. Toxocara canis was most prevalent in urban stray dogs (75·0%) and least prevalent in foxes from the montane locality (7·0%). The prevalence of Toxocara cati in cats at the urban, rural and montane localities was 66·2, 65·2 and 76·9%, respectively. In clinically healthy human populations, the highest seroprevalence was detected in the rural locality (14·0%). Children of the same area were 3 times more seropositive (12·9%) than those from the urban and montane localities (4·3 and 4·0%). Our studies suggest an important role for small mammals as paratenic hosts – reservoirs of Toxocara larvae – in maintaining toxocariasis foci. In this respect toxocariasis may be classified as an anthropopurgic focal zoonosis.

Hosts living on islands carry few parasite species, and the prevalence and intensity of directly transmitted parasites are often higher in insular than in mainland populations. However, it is unclear whether density or other features of insular populations can be responsible for the pattern observed. We compared the parasite richness, prevalence and intensity of parasites between 2 feral populations of cats living either at low density on an island (Kerguelen) or at high density on the mainland (Lyon). Parasite richness was higher in Lyon than in Kerguelen, where only Toxocara cati was found. T. cati egg prevalence was higher in Kerguelen (71·1%) than in Lyon (58·0%). Because cat density cannot explain this pattern, we propose that the low number of parasite species, the diet and/or immunity of cats act to increase prevalence in Kerguelen. Moreover, prevalence, intensity and variance-to-mean ratio increased with age and body mass in Kerguelen whereas, in Lyon, prevalence decreased with age and body mass. We hypothesize that the pattern of exposure differs between populations, and that density-dependent parasite mortality is lower in Kerguelen than in Lyon. We discuss the consequences concerning the influence of parasites on insular host populations.