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IntroductionToxoplasma gondii, responsible for causing toxoplasmosis, is a prevalent food and waterborne pathogen worldwide. It commonly infects warm-blooded animals and affects more than a third of the global human population. Once ingested, the parasite enters the host’s small intestine and rapidly disseminates throughout the body via the bloodstream, infiltrating various tissues. Leukocyte-driven responses are vital against T. gondii, with neutrophils playing a dual role: swiftly recruited to infection sites, releasing inflammatory mediators, and serving as a replication hub and Trojan horses, aiding parasite spread. Neutrophils from various hosts release extracellular traps (NETs) against the protozoan. However, gaps persist regarding the mechanisms of NETs production to parasite and their significance in infection control. This study investigates the interplay between human neutrophils and T. gondii, exploring dynamics, key molecules, and signaling pathways involved in NETs production upon protozoan challenge.Methods and ResultsUsing confocal and electron microscopy, live cell imaging, pharmacological inhibitors, and DNA quantification assays, we find that human neutrophils promptly release both classical and rapid NETs upon pathogen stimulation. The NETs structure exhibits diverse phenotypes over time and is consistently associated with microorganisms. Mechanisms involve neutrophil elastase and peptidylarginine deiminase, along with intracellular calcium signaling and the PI3K pathway. Unexpectedly, human traps do not diminish viability or infectivity, but potentially aid in capturing parasites for subsequent neutrophil phagocytosis and elimination.DiscussionBy revealing NETs formation mechanisms and their nuanced impact on T. gondii infection dynamics, our findings contribute to broader insights into host-pathogen relationships.

Toxoplasma gondii is the causative agent of toxoplasmosis, a disease that affects warm-blooded animals and one third of the human population worldwide. Pregnant women who have never been exposed to the parasite constitute an important risk group, as infection during pregnancy often leads to congenital toxoplasmosis, the most severe form of the disease. Current therapy for toxoplasmosis is the same as it was 50 years ago and has little or no effect when vertical transmission occurs. Therefore, it is urgent to develop new strategies to prevent mother-to-fetus transmission. The implementation of experimental animal models of congenital toxoplasmosis that reproduces the transmission rates and clinical signs in humans opens an avenue of possibilities to interfere in the progression of the disease. In addition, knowing the parasite load in maternal and fetal tissues after infection, which may be related to organ abnormalities and disease outcome, is another important step in designing a promising intervention strategy. Therefore, we implemented here a murine model of congenital toxoplasmosis with outbred Swiss Webster mice infected intravenously with tachyzoites of the ME49 strain of T. gondii that mimics the frequency of transmission of the parasite, as well as important clinical signs of human congenital toxoplasmosis, such as macrocephaly, in addition to providing a highly sensitive quantitative real-time PCR assay to assess parasite load in mouse tissues. As the disease is not restricted to humans, also affecting several domestic animals, including companion animals and livestock, they can also benefit from the model presented in this study.

The life cycles of many parasitic nematodes include terrestrial gastropods as intermediate hosts. Over the past few decades, a number of cases of parasitism between molluscs and medically-important nematodes have been reported in Brazil, in particular, those involving the invasive giant African gastropod, Achatina fulica, and zoonoses caused by the nematodes Angiostrongylus cantonensis and Angiostrongylus costaricensis, the etiological agents of neuroangiostrongyliasis and abdominal angiostrongyliasis, respectively. In the present study, larvae found infecting A. fulica, Latipes erinaceus, and Thaumastus taunaisii, from two localities in the Brazilian state of Rio de Janeiro were characterized using light and scanning electron microscopy, and sequences of the 18S rRNA and MT-CO1 genes. Genetic markers allowed to identify the larvae collected in the present study as Cruzia tentaculata, whose adults parasitize didelphid marsupials in the Americas. These findings indicate that both native and non-native gastropods may act as intermediate hosts and represent a previously unnoticed heteroxenous life cycle of C. tentaculata.

Abstract The Japanese invasive jumping snail Ovachlamys fulgens is a pest of ornamental plants and an intermediate host of a nematode that causes eosinophilic meningitis. We expand its distribution to eight municipalities from Rio de Janeiro State, and one locality from the Paraná State, and generated for the first time partial sequences of the cytochrome c oxidase subunit I (COI) gene for Brazilian populations. External morphology, reproductive system, shell, radula, and jaw were also analyzed and described. Twenty-one lots were collected from Rio de Janeiro, Niterói, Magé, Miguel Pereira, Petrópolis, Teresópolis, Nova Friburgo, Bom Jardim and Paraty, in Rio de Janeiro State, and from Foz do Iguaçu, Paraná State. External morphology, shell and reproductive system were typical of O. fulgens, with some peculiarities found in the shell and radula. A single haplotype was found, which was 100% similar to sequences of COI available in GenBank for specimens from Japan and Argentina. The species seems to be adapted to many habitats and be rapidly expanding its distribution in Southeastern and Southern Brazil, and other South America countries. We highlight the importance of monitoring O. fulgens, considering its potential to compete with native mollusks, attack several plants, and be a transmitter of diseases.

Toxoplasma gondii is the causative agent of toxoplasmosis, an infectious disease that affects over 30% of the human world population, causing fatal infections in immunocompromised individuals and neonates. The life cycle of T. gondii is complex, and involves intermediate hosts (birds and mammals) and definitive hosts (felines, including domestic cats). The innate immune repertoire against the parasite involves the production of neutrophil extracellular traps (NET), and neutrophils from several intermediate hosts produce NET induced by T. gondii. However, the mechanisms underlying NET release in response to the parasite have been poorly explored. Therefore, the aims of this study were to investigate whether neutrophils from cats produce NET triggered by T. gondii and to understand the mechanisms thereby involved. Neutrophils from cats were stimulated with T. gondii tachyzoites and NET-derived DNA in the supernatant was quantified during the time. The presence of histone H1 and myeloperoxidase was detected by immunofluorescence. We observed that cat neutrophils produce both classical and rapid/early NET stimulated by T. gondii. Inhibition of elastase, intracellular calcium, and phosphatidylinositol 3-kinase (PI3K)-δ partially blocked classical NET release in response to the parasite. Electron microscopy revealed strands and networks of DNA in close contact or completely entrapping parasites. Live imaging showed that tachyzoites are killed by NET. We conclude that the production of NET is a conserved strategy to control infection by T. gondii amongst intermediate and definitive hosts.

Os autores descreveram as características morfológicas de Triatoma klugi Carcavallo, Jurberg, Lent & Galvão, 2001 pertencente ao grupo de espécies que compõem o "complexo T. oliveirai". Até o presente tem sido difícil separar essas espécies com base nas características ninfais, o que justifica o desenvolvimento deste trabalho. Os espécimes foram coletados em frestas de rochas no morro Malavok na localidade de Linha Brasil, município de Nova Petrópolis, no Estado do Rio Grande do Sul. O local de captura dos triatomíneos situa-se entre 700 e 800 m de altitude (29º18'38''S, 51º04'57''W). Juntamente com T. oliveirai (Neiva, Pinto & Lent, 1939), espécie morfologicamente mais próxima, são as únicas do complexo que não foram encontradas, até o momento, no Estado do Mato Grosso, Mato Grosso do Sul e Goiás, Brasil. Em condições experimentais, já foi testada e comprovada a sua susceptibilidade à infecção pelo Trypanosoma cruzi (Chagas, 1909) e Trypanosoma rangeli (Tejera, 1920). A análise comparativa das ornamentações do exocório dos ovos e de três estádios ninfais de T. klugi por microscopia óptica e microscopia eletrônica de varredura mostrou algumas particularidades morfológicas, com destaque para: a face ventral da cabeça, o sulco estridulatório e os últimos segmentos abdominais (IX, X - futura genitália e XI - tubo anal). Esses dados contribuem para a ampliação dos parâmetros diferenciais visando à diagnose de T. klugi durante o seu desenvolvimento ninfal.The authors had described the morphologic characteristics of Triatoma klugi Carcavallo, Jurberg, Lent & Galvão, 2001 to the group of species belonging to "T. oliveirai complex". Until the moment it has been difficult to separate on the nymphs characteristics basis these species, what it justifies the development of this work. The specimens had been collected in openings of rocks in the Malavok mount in the locality of Linha Brazil, city of Nova Petrópolis, in the State of Rio Grande do Sul. The local of capture of the Triatominae is placed enters 700 and 800 m of altitud (29º18'38''S, 51º04'57''W). Together with T. oliveirai (Neiva, Pinto & Lent, 1939), its morphologically related species, is the only ones of the complex that had not been found, until the moment, in the State of Mato Grosso, Mato Grosso do Sul and Goiás, Brazil. In experimental conditions, already it was tested and proven its susceptibilities to the infection for Trypanosoma cruzi (Chagas, 1909) and Trypanosoma rangeli (Tejera, 1920). The comparative analysis of the ornaments of the exocorium of eggs and of three nymphs of T. klugi by optic microscopy and scanning electron microscopy showed some morphologic particularities, with prominence for: the ventral face of the head, the stridulatory sulcus and the last abdominal segments (IX, X - future genitalia and XI - anal duct). These data contribute for the magnifying of the distinguishing parameters aiming at diagnosis of T. klugi during its nymphal development.

Macrophages and muscle cells are the main targets for invasion of Trypanosoma cruzi. Ultrastructural studies of this phenomenon in vitro showed that invasion occurs by endocytosis, with attachment and internalization being mediated by different components capable of recognizing epi-or trypomastigotes (TRY). A parasitophorus vacuole was formed in both cell types, thereafter fusing with lysosomes. Then, the mechanism of T. cruzi invasion of host cells (HC) is essentially similar (during a primary infection in the abscence of a specific immune response), regardless of wether the target cell is a professional or a non-professional phagocytic cell. Using sugars, lectins, glycosidases, proteinases and proteinase inhibitors, we observed that the relative balance between exposed sialic acid and galactose/N-acetyl galactosamine (GAL) residues on the TRY surface, determines the parasite's capacity to invade HC, and that lectin-mediated phagocytosis with GAL specificity is important for internalization of T. cruzi into macrophages. On the other hand, GAL on the surface to heart muscle cells participate on TRY adhesion. TRY need to process proteolytically both the HC and their own surface, to expose the necessary ligands and receptors that allow binding to, and internalization in the host cell. The diverse range of molecular mechanisms which the parasite could use to invade the host cell may correspond to differences in the available "receptors"on the surface of each specific cell type. Acute phase components, with lectin or proteinase inhibitory activities (a-macroglobulins), may also be involved in T. cruzi-host cell interaction.

Abstract Background The interest in the mechanisms involved in Toxoplasma gondii lipid acquisition has steadily increased during the past few decades, but it remains not completely understood. Here, we investigated the biogenesis and the fate of lipid droplets (LD) of skeletal muscle cells (SkMC) during their interaction with T. gondii by confocal and electron microscopy. We also evaluated whether infected SkMC modulates the production of prostaglandin E2 (PGE2), cytokines interleukin-12 (IL-12) and interferon-gamma (INF-g), and also the cyclooxygenase-2 (COX-2) gene induction. Methods Primary culture of skeletal muscle cells were infected with tachyzoites of T. gondii and analysed by confocal microscopy for observation of LD. Ultrastructural cytochemistry was also used for lipid and sarcoplasmatic reticulum (SR) detection. Dosage of cytokines (IL-12 and INF-g) by ELISA technique and enzyme-linked immunoassay (EIA) for PGE2 measurement were employed. The COX-2 gene expression analysis was performed by real time reverse transcriptase polymerase chain reaction (qRT-PCR). Results We demonstrated that T. gondii infection of SkMC leads to increase in LD number and area in a time course dependent manner. Moreover, the ultrastructural analysis demonstrated that SR and LD are in direct contact with parasitophorous vacuole membrane (PVM), within the vacuolar matrix, around it and interacting directly with the membrane of parasite, indicating that LD are recruited and deliver their content inside the parasitophorous vacuole (PV) in T. gondii-infected SkMC. We also observed a positive modulation of the production of IL-12 and IFN-g, increase of COX-2 mRNA levels in the first hour of T. gondii-SkMC interaction and an increase of prostaglandin E2 (PGE2) synthesis from 6 h up to 48 h of infection. Conclusions Taken together, the close association between SR and LD with PV could represent a source of lipids as well as other nutrients for the parasite survival, and together with the increased levels of IL-12, INF-g and inflammatory indicators PGE2 and COX-2 might contribute to the establishment and maintenance of chronic phase of the T. gondii infection in muscle cell.

The present paper summarizes new approaches regarding the progress done to the understanding of the interaction of Trypanosoma cruzi-cardiomyocytes. Mannose receptors localized at the surface of heart muscle cell are involved in binding and uptake of the parasite. One of the most striking events in the parasite-heart muscle cells interaction is the disruption of the actin cytoskeleton. We have investigated the regulation of the actin mRNA during the cytopathology induced in myocardial cells by the parasite. T. cruzi invasion increases calcium resting levels in cardiomyocytes. We have previously shown that Ca2+ ATPase of the sarcoplasmic reticulum (SERCA) is involved in the invasion of T. cruzi in cardiomyocytes. Treating the cells with thapsigargin, a drug that binds to all SERCA ATPases and causes depletion of intracellular calcium stores, we found a 75% inhibition in the T. cruzi-cardiomyocytes invasion.