Toxocara canis infection worsens the course of experimental autoimmune encephalomyelitis in mice.

Toxocara canis, a gastrointestinal parasite of canids, is also highly prevalent in many paratenic hosts, such as mice and humans. As with many other helminths, the infection is associated with immunomodulatory effects, which could affect other inflammatory conditions including autoimmune and allergic diseases. Here, we investigated the effect of T. canis infection on the course of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. Mice infected with 2 doses of 100 T. canis L3 larvae 5 weeks prior to EAE induction (the Tc+EAE group) showed higher EAE clinical scores and greater weight loss compared to the non-infected group with induced EAE (the EAE group). Elevated concentrations of all measured serum cytokines (IL-1α, IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ and TNF-α) were observed in the Tc+EAE group compared to the EAE group. In the CNS, the similar number of regulatory T cells (Tregs; CD4+FoxP3+Helios+) but their decreased proportion from total CD4+ cells was found in the Tc+EAE group compared to the EAE group. This could indicate that the group Tc+EAE harboured significantly more CD4+ T cells of non-Treg phenotype within the affected CNS. Altogether, our results demonstrate that infection of mice with T. canis worsens the course of subsequently induced EAE. Further studies are, therefore, urgently needed to reveal the underlying pathological mechanisms and to investigate possible risks for the human population, in which exposure to T. canis is frequent.

Antigenic Proteins from the Excretory-Secretory Products of Toxocara canis Larvae and Evaluation of Their Potential for Immunodiagnostics of Larval Toxocarosis.

BACKGROUND: Larval toxocarosis is a zoonosis caused by larvae of Toxocara canis and T. cati, a gastrointestinal nematode of canids and felids, respectively. Diagnosis is usually performed by ELISA IgG using Toxocara excretory-secretory products as an antigen. Due to laboriousness of isolation of the products and subsequent process of standardization of antigenic compounds, routine use of this method is limited and can produce inaccurate diagnostical results. The purpose of this study was to discover new specific antigenic proteins that could be used in routine serological methods of larval toxocarosis. MATERIALS AND METHODS: Toxocara excretory-secretory products were collected and separated by SDS-PAGE. Proteins from the gel were electro-transferred to a membrane and incubated with mouse sera. Antigenic proteins were analyzed using the liquid chromatography-tandem mass spectrometry approach. Selected proteins were prepared in recombinant form and tested with mice and human sera by ELISA and Western blot. RESULTS: A total of four recombinant protein antigens were prepared (rTc-TES-26, rTc-ASA, rTc-PDP, and rTc-ASP). They were analyzed by ELISA and Western blot using mice and human sera. For all sera, three of the four recombinant antigens correlated with Toxocara excretory-secretory products in ELISA analysis. By Western blot, the infection was confirmed in all experimentally infected mice and two out of seven human patients. CONCLUSION: Combination of the presented methods and analyses represents a possible method of effective identification of Toxocara protein antigens for the purpose of routine serodiagnosis.

Seroprevalence of Larval Toxocarosis in the Czech Republic.

BACKGROUND: Larval toxocarosis (LT), a zoonotic disease transmitted by dogs, cats, and other carnivores, is caused by roundworms of the genus Toxocara. Humans become infected by ingesting embryonated eggs of this parasite. In this study, we present data on the seroprevalence of LT in the Czech Republic collected by the National Reference Laboratory for Tissue Helminthoses in 2012-2016. METHODS: Using enzyme-linked immunosorbent assay, a total of 4428 adults and children with or without clinical symptoms were examined for the presence of IgG antibodies against Toxocara canis excretory-secretory antigens. RESULTS: Of all the persons examined, specific Toxocara antibodies were detected in 160 (3.6%) individuals. There were, however, significant differences between various regions, with seropositivity rates ranging from 1.4 to 7.5%. CONCLUSION: In comparison to studies from 1998 and 2004, our results suggest a decrease in overall Toxocara seroprevalence in the Czech population, whereby the rates are similar to or even lower than rates in some other Central European countries.

Heterolobosean amoebae from Arctic and Antarctic extremes: 18 novel strains of Allovahlkampfia, Vahlkampfia and Naegleria.

The diversity of heterolobosean amoebae, important members of soil, marine and freshwater microeukaryote communities in the temperate zones, is greatly under-explored in high latitudes. To address this imbalance, we studied the diversity of this group of free-living amoebae in the Arctic and the Antarctic using culture dependent methods. Eighteen strain representatives of three heterolobosean genera, Allovahlkampfia Walochnik et Mulec, 2009 (1 strain), Vahlkampfia Chatton et Lalung-Bonnaier, 1912 (2) and Naegleria Alexeieff, 1912 (15) were isolated from 179 samples of wet soil and fresh water with sediments collected in 6 localities. The Allovahkampfia strain is the first representative of the genus from the Antarctic; 14 strains (7 from the Arctic, 7 from the Antarctic) of the highly represented genus Naegleria complete the 'polar' cluster of five Naegleria species previously known from the Arctic and Sub-Antarctic regions, whereas one strain enriches the 'dobsoni' cluster of Naegleria strains of diverse origin. Present isolations of Naegleria polarisDe Jonckheere, 2006 from Svalbard, in the Arctic and Vega Island, in the Antarctic and N. neopolarisDe Jonckheere, 2006 from Svalbard and Greenland in the Arctic, and James Ross Island, the Antarctic demonstrate their bipolar distribution, which in free-living amoebae has so far only been known for Vermistella Morand et Anderson, 2007.