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Human toxocariasis is a neglected anthropozoonosis with global distribution. Treatment is based on the administration of anthelmintics; however, their effectiveness at the tissue level is low to moderate, necessitating the discovery of new drug candidates. Several groups of synthetic compounds, including coumarin derivatives, have demonstrated bioactivity against fungi, bacteria, and even parasites, such as Dactylogyrus intermedius, Leishmania major, and Plasmodium falciparum. The aim of this study was to evaluate the effect of ten coumarin-derived compounds against Toxocara canis larvae using in vitro, cytotoxicity, and in silico tests for selecting new drug candidates for preclinical tests aimed at evaluating the treatment of visceral toxocariasis. The compounds were tested in vitro in duplicate at a concentration of 1 mg/mL, and compounds with larvicidal activity were serially diluted to obtain concentrations of 0.5 mg/mL; 0.25 mg/mL; 0.125 mg/mL; and 0.05 mg/mL. The tests were performed in a microculture plate containing 100 T. canis larvae in RPMI-1640 medium. One compound (COU 9) was selected for cytotoxicity analysis using J774.A1 murine macrophages and it was found to be non-cytotoxic at any concentration tested. The in silico analysis was performed using computational models; the compound presented adequate results of oral bioavailability. To confirm the non-viability of the larvae, the contents of the microplate wells of COU 9 were inoculated intraperitoneally (IP) into female Swiss mice at 7-8 weeks of age. This confirmed the larvicidal activity of this compound. These results show that COU 9 exhibited larvicidal activity against T. canis larvae, which, after exposure to the compound, were non-viable, and that COU 9 inhibited infection in a murine model. In addition, COU 9 did not exhibit cytotoxicity and presented adequate bioavailability in silico, similar to albendazole, an anthelmintic, which is the first choice for treatment of human toxocariasis, supporting the potential for future investigations and preclinical tests on COU 9., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Human toxocariasis is a parasitic anthropozoonosis that is difficult to treat and control. A previous study carried out with Lactobacillus acidophilus ATCC 4356 revealed that the cell free supernatant (CFS) of this probiotic killed 100% of Toxocara canis larvae in vitro. The present study aimed to investigate the characteristics of the CFS of L. acidophilus ATCC 4356, which may be involved in its larvicidal effects on T. canis. L. acidophilus ATCC 4356 was cultured, and lactic and acetic acids present in the CFS were quantified by high performance liquid chromatography (HPLC). The levels of pH and H 2 O 2 were also analyzed. To assess the larvicidal effect of the CFS, this was tested pure and diluted (1:2 to 1:128) on T. canis larvae. High concentrations of lactic and acetic acids were detected in the CFS. The acidity of the pure CFS was observed at pH 3.8, remaining acidic at dilutions of 1:2 to 1:16. Regarding the in vitro larvicidal effect, 100% death of T. canis larvae was observed using the pure CFS and 1:2 dilution. Based on these results, it can be inferred that the presence of higher concentrations of organic acids and low pH of the medium contributed to the larvicidal activity of the CFS of L. acidophilus ATCC 4356. In addition, the maintenance of the larvicidal effect, even after dilution, suggests a greater chance of the larvicidal effect of this CFS against T. canis in vivo., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Toxocariasis is an anthropozoonosis caused by the helminth Toxocara canis that shows different clinical manifestations as visceral, ocular, or neurological toxocariasis forms. Probiotics have been studied as alternatives to prevent and treat this parasitosis. Lactobacillus rhamnosus is a prospect that presents immunomodulatory activity that acts to strengthen the intestinal barrier. In this context, the main objective of this study was to evaluate the protective capacity and immunomodulatory action of the probiotic Lactobacillus rhamnosus at the level of the intestinal mucosa in different stages of T. canis infection (acute and chronic). Mice were supplemented by oral gavage with 1 × 10 7 UFC/mL L. rhamnosus for 15 days before inoculation with 100 embryonated eggs of T. canis. Euthanasia of mice was conducted at three different time points: 2, 15 and 30 days post-inoculation (PI). The brain, lungs and liver were collected to evaluate the intensity of infection. The small intestines were removed, and mucosal cells of the duodenum were collected to perform gene analysis of IFN-γ, IL-10, IL-4 and IL-13 by real-time polymerase chain reaction (qPCR). Jejunum and ileum segments were analysed by histological techniques. A reduction of 51% in infection intensity was observed in the tissue of supplemented animals evaluated 2 days PI; however, analysis of groups 15 and 30 days PI did not show a protective effect. The intestinal mucosa of supplemented animals presented an inflammatory process that initiated at 2 days PI, persisted at 15 days PI and had regressed at 30 days PI. IL-13 transcription was increased in the probiotic group 2 days after supplementation ended; however, the same increase was not observed in the group that was supplemented and infected. Toxocara canis modulated the local immune system, with suppression of IFN-γ at 2 days PI and increased levels of IL-4 and IL-10 at 15 days PI. These results indicate that, under the studied conditions, the protective effect of Lactobacillus rhamnosus against infection caused by T. canis is not related to IL-4, IL-10 or IFN-γ but could be influenced by IL-13 action at 2 days PI. The probiotic stimulated immune cell recruitment to the intestinal mucosa, which can be involved in the diminished capacity of larval penetration in the mucosa, resulting in the reduced infection intensity observed during acute infection., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Funding agency: National Council for Scientific and Technological Development, Brazil., (Copyright © 2023 Elsevier GmbH. All rights reserved.)

Experimental studies have demonstrated the potential of probiotics to control visceral toxocariasis, which is a tissue parasitosis that is difficult to treat. This study evaluated the in vitro activity of probiotics and their supernatants on Toxocara canis larvae. The probiotics Lactobacillus rhamnosus (ATCC 7469), Lactobacillus paracasei (ATCC 335), Saccharomyces boulardii, Saccharomyces cerevisiae, and Bacillus cereus var. toyoi were tested in the following preparations: probiotic (P) 1 × 102 to 1 × 109 colony-forming units (CFUs), inactivated probiotic (IP) 1 × 102 to 1 × 109 CFUs, supernatant probiotic (SUpP), and inactivated probiotic supernatant (SupIP). The probiotics and their respective supernatants were separately incubated with 100 T. canis larvae per well using microculture plates with RPMI-1640 medium for 48 hr at 37 C and 5% CO2. The evaluation of the in vitro tests was based on the viability of T. canis larvae, through morphologic integrity, positive motility, and the absence of trypan blue stain. Only culture supernatants (SUpP and SUpIP) of Lactobacillus spp. resulted in 100% dead larvae, whereas S. boulardii showed larvicidal activity in T. canis >70%. The rest of the tests did not show larvicide activity. Therefore, it is important to investigate the supernatant effects of Lactobacillus spp. and S. boulardii in vivo on T. canis visceral infections, their mechanisms of action, and major metabolites involved., (© American Society of Parasitologists 2023.)