IL-17RA receptor signaling contributes to lung inflammation and parasite burden during Toxocara canis infection in mice.

IL-17 is a cytokine produced by innate and acquired immunity cells that have an action against fungi and bacteria. However, its action in helminth infections is unclear, including in Toxocara canis infection. Toxocariasis is a neglected zoonosis representing a significant public health problem with an estimated seroprevalence of 19% worldwide. In the present study, we describe the immunopathological action of IL-17RA in acute T. canis infection. C57BL/6j (WT) and IL-17RA receptor knockout (IL-17RA-/-) mice were infected with 1000 T. canis eggs. Mice were evaluated 3 days post-infection for parasite load and white blood cell count. Lung tissue was harvested for histopathology and cytokine expression. In addition, we performed multiparametric flow cytometry in the BAL and peripheral blood, evaluating phenotypic and functional changes in myeloid and lymphoid populations. We showed that IL-17RA is essential to control larvae load in the lung; however, IL-17RA contributed to pulmonary inflammation, inducing inflammatory nodular aggregates formation and presented higher pulmonary IL-6 levels. The absence of IL-17RA was associated with a higher frequency of neutrophils as a source of IL-4 in BAL, while in the presence of IL-17RA, mice display a higher frequency of alveolar macrophages expressing the same cytokine. Taken together, this study indicates that neutrophils may be an important source of IL-4 in the lungs during T. canis infection. Furthermore, IL-17/IL-17RA axis is important to control parasite load, however, its presence triggers lung inflammation that can lead to tissue damage.

Prevalence of Toxocariasis and Its Risk Factors in Patients with Eosinophilia in Korea.

Eosinophilia occurs commonly in many diseases including allergic diseases and helminthic infections. Toxocariasis has been suggested as one cause of eosinophilia. The present study was undertaken to examine the prevalence of toxocariasis in patients with eosinophilia and to identify the risk factors for toxocariasis. This prospective cohort study recruited a total of 81 patients with eosinophilia (34 males and 47 females) who visited the outpatient clinic at Seoul National University Hospital from January 2017 to February 2018 and agreed to participate in this study. The prevalence of toxocariasis was examined by T. canis-specific ELISA, and the various risk factors for toxocariasis were evaluated by a questionnaire survey. Among 81 patients with eosinophilia, 18 were positive for anti-T. canis antibodies (22.2%); 88.9% were male (16/18) and 11.1% were female (2/18). Multivariate statistical analysis revealed that males (OR 21.876, 95% CI: 1.667-287.144) with a history of consuming the raw meat or livers of animals (OR 5.899, 95% CI: 1.004-34.669) and a heavy alcohol-drinking habit (OR 8.767, 95% CI: 1.018-75.497) were at higher risk of toxocariasis in patients with eosinophilia. Toxocariasis should be considered a potential cause of eosinophilia when the patient has a history of eating the raw meat or livers of animals in Korea. A single course of albendazole is recommended to reduce the migration of Toxocara larvae in serologically positive cases with eosinophilia.

Urticaria and silent parasitism by Ascaridoidea: Component-resolved diagnosis reinforces the significance of this association.

Urticaria remains a major problem in terms of aetiology, investigation, and management, and although parasitic diseases are considered potential causes, the absence of a consistent link between parasitic infections and skin allergy symptoms leads to the need for a deeper study of parameters that support this association. The objectives of this study were to analyse a possible relationship between parasitism by Ascarididae (Toxocara canis and Anisakis simplex) and the clinical expression of urticaria and to identify possible parasitic molecular markers for improving the diagnosis of unknown urticaria aetiology. The prevalence of Toxocara and Anisakis infestations was evaluated by measuring the levels of specific IgG (sIgG) and IgE (sIgE) antibodies against crude extracts and isolated components from whole larvae of Anisakis simplex (Ani s 1, Ani s 3 and Ani s 7) and Toxocara canis (TES-120, TES-70, TES-32 and TES-26) using immunologic and molecular diagnostic methods. A cross-sectional study was performed in a group of 400 individuals. The study group consisted of 95 patients diagnosed with urticaria (55 with chronic urticaria and 40 with acute urticaria). A control group consisted of 305 subjects without urticaria (182 diagnosed with respiratory allergy and 123 without allergy). Statistically significant differences were demonstrated in the seroprevalence of specific IgG and IgE antibodies between the urticaria patients and the healthy general population when isolated ascarid antigens were evaluated. The prevalence of IgG antibodies against Ani s 1, IgE antibodies against TES-120 and IgE antibodies against TES-70 were significantly different between the control individuals (healthy general population) and patients with urticaria. Moreover, the urticaria patient group demonstrated a higher seroprevalence of antibodies (sIgE and sIgG) against Anisakis simplex larva whole extract than the control group but just with statistically diferences when sIgE was evaluated. The presence of IgE and/or IgG antibodies against Ani s 3 (tropomyosin) can help to discriminate between patients with and without urticaria. Both ascarids seem to be associated with urticaria, although in our region, Anisakis seems to have greater involvement than Toxocara in this relationship. Molecular diagnostics can be used to associate urticaria with parasite infestations. Tropomyosin and Ani s 1 were the most relevant markers to demonstrate the association between urticaria and the most relevant Ascarididae parasites in our region.

Prevalence of Toxocara and Toxascaris infection among human and animals in Iran with meta-analysis approach.

BACKGROUND: Toxocariasis is a worldwide zoonotic parasitic disease caused by species of Toxocara and Toxascaris, common in dogs and cats. Herein, a meta-analysis was contrived to assess the prevalence of Toxocara/Toxascaris in carnivore and human hosts in different regions of Iran from April 1969 to June 2019. METHODS: The available online articles of English (PubMed, Science Direct, Scopus, and Ovid) and Persian (SID, Iran Medex, Magiran, and Iran Doc) databases and also the articles that presented in held parasitology congresses of Iran were involved. RESULTS: The weighted prevalence of Toxocara/Toxascaris in dogs (Canis familiaris) and cats (Felis catus) was 24.2% (95% CI: 18.0-31.0%) and 32.6% (95% CI: 22.6-43.4%), respectively. Also, pooled prevalence in jackal (Canis aureus) and red fox (Vulpes vulpes) was 23.3% (95% CI: 7.7-43.2%) and 69.4% (95% CI: 60.3-77.8%), correspondingly. Weighted mean prevalence of human cases with overall 28 records was 9.3% (95% CI: 6.3-13.1%). The weighted prevalence of Toxocara canis, Toxocara cati, and Toxascaris leonina was represented as 13.8% (95% CI: 9.8-18.3%), 28.5% (95% CI: 20-37.7%) and 14.3% (95% CI: 8.1-22.0%), respectively. CONCLUSION: Our meta-analysis results illustrate a considerable prevalence rate of Toxocara/Toxascaris, particularly in cats and dogs of northern parts of Iran. The presence of suitable animal hosts, optimum climate and close contact of humans and animals would have been the reason for higher seroprevalence rates of human cases in our region. Given the significance clinical outcomes of human Toxocara/Toxascaris, necessary measures should be taken.

Electrochemical microfluidic immunosensor based on TES-AuNPs@Fe3O4 and CMK-8 for IgG anti-Toxocara canis determination.

We report a microfluidic immunosensor for the electrochemical determination of IgG antibodies anti-Toxocara canis (IgG anti-T. canis). In order to improve the selectivity and sensitivity of the sensor, core-shell gold-ferric oxide nanoparticles (AuNPs@Fe3O4), and ordered mesoporous carbon (CMK-8) in chitosan (CH) were used. IgG anti-T. canis antibodies detection was carried out using a non-competitive immunoassay, in which excretory secretory antigens from T. canis second-stage larvae (TES) were covalently immobilized on AuNPs@Fe3O4. CMK-8-CH and AuNPs@Fe3O4 were characterized by transmission electron microscopy, scanning electron microscopy, energy dispersive spectrometry, cyclic voltammetry, electrochemical impedance spectroscopy, and N2 adsorption-desorption isotherms. Antibodies present in serum samples immunologically reacted with TES, and then were quantified by using a second antibody labeled with horseradish peroxidase (HRP-anti-IgG). HRP catalyzes the reduction from H2O2 to H2O with the subsequent oxidation of catechol (H2Q) to p-benzoquinone (Q). The enzymatic product was detected electrochemically at _100 mV on a modified sputtered gold electrode. The detection limit was 0.10 ng mL-1, and the coefficients of intra- and inter-assay variation were less than 6%, with a total assay time of 20 min. As can be seen, the electrochemical immunosensor is a useful tool for in situ IgG antibodies anti-T. canis determination.

Immune response to chronic Toxocara canis infection in a mice model.

AIMS: The zoonotic nematode Toxocara canis causes larva migrans syndrome that induces an immune response characterized by the production of antibodies and eosinophilia. A Th2 polarization has been associated with the infection, but there are still details of the cellular and humoral immune response that need to be described. Thus, the aim of this study was to describe the systemic host immune response to T canis chronic infection in a mouse model. METHODS AND RESULTS: BALB/c mice were inoculated once with 500 T canis embryonated eggs, per os. After 49 days, the amounts of larval found in brain and muscle tissues were statistically two and four times higher, respectively, than the amounts found in lung, liver, kidney or heart tissues. Splenic proportions of F4/80+ cells, as well as B, cytotoxic T and CD4+ Foxp3+ lymphocytes, were statistically higher (P ≤ .05, P ≤ .01, P ≤ .001 and P ≤ .001, respectively) as compared with control mice. In lymph nodes, some of these proportions changed, with the exception of F4/80+ cells. IgG1 levels in infected mice sera were increased. IL-4, IL-10 and VEGF levels were statistically higher in spleen (P ≤ .05, all) and sera (P ≤ .01, P ≤ .05 and P ≤ .05, respectively) in the infected mice. Also, in infected animals, IL-5 serum levels were increased (P ≤ .01). CONCLUSION: These results suggest that T canis chronic infection in BALB/c mice results in a type 2 response with an incipient regulatory response.

Cytokine production and signalling in human THP-1 macrophages is dependent on Toxocara canis glycans.

The effect of Toxocara canis antigens on cytokine production by human THP-1 macrophages was studied in vitro. Toxocara Excretory-Secretory products (TES) and recombinant mucins (Tc-MUC-2, Tc-MUC-3, Tc-MUC-4, and Tc-MUC-5) as well as deglycosylated forms of these antigens were used in the study. TES products stimulated macrophages to produce the innate proinflammatory IL-1ß, IL-6, and TNF-α cytokines regardless of the presence of glycans. Recombinant mucins induced glycan-dependent cytokine production. Sugar moieties led to at least 3-fold higher production of regulatory IL-10 as well as proinflammatory cytokines. The presence of glycans on mucins also affected the downstream signalling pathways in stimulated cells. The most prominent difference was noted in AKT and AMPK kinase activation. AKT phosphorylation was observed in cells stimulated with glycosylated mucins, whereas treatment with deglycosylated antigens led to AMPK phosphorylation. MAP kinase family members such as JNK and p38 and c-Jun transcription factor were phosphorylated in both cases what suggests that toll-like receptor signalling may be involved in mucin-treated macrophages. This pathway is however modified by other signalling molecules as only mucins containing intact sugars significantly induced the production of cytokines.

Histopathological characterization of Toxocara canis- and T. cati-induced neurotoxocarosis in the mouse model.

Infective larvae of Toxocara canis and T. cati, the common roundworms of dogs and cats, may invade the central nervous system of paratenic hosts, including humans, causing neurotoxocarosis (NT). Previous studies on NT in the model organism "mouse" have indicated distinct differences between T. canis and T. cati regarding larval migration patterns as well as the severity of clinical symptoms and behavioural alterations. The objective of the present study was to provide an extensive characterization of the underlying histopathological alterations, comparing T. canis- and T. cati-induced changes in different brain areas over the course of murine infection. Four histological sections of five brains each of T. canis- and T. cati-infected as well as uninfected C57Bl/6 mice were investigated 7, 14, 28, 42, 70 and 98 days post infection (dpi), while brains of T. cati-infected and control mice were also available 120 and 150 dpi. In addition to haematoxylin-eosin and luxol fast blue-cresyl violet staining, immunohistochemistry was employed to study microglia/macrophage cell morphology and to detect accumulation of ß-amyloid precursor protein (ß-APP) as an indicator of axonal damage. Haemorrhages, eosinophilic vasculitis and activated microglia/macrophages were detected in both infection groups starting 7 dpi, followed by eosinophilic meningitis in cerebra as from 14 dpi. Overall, little differences in the proportion of animals affected by these alterations were found between the two infection groups. In contrast, the proportion of animals displaying ß-APP accumulation was significantly higher in the T. canis than T. cati group as from 28 dpi regarding the cerebrum as well as at 98 dpi regarding the cerebellum. In T. canis-infected mice, myelinophagic microglia/macrophages ("gitter cells") appeared as from 14 dpi, whereas these were first observed at 70 dpi in T. cati-infected animals. The proportion of animals displaying demyelination and/or gitter cells in the cerebrum was significantly higher in the T. canis than T. cati group as from 28 dpi, and at 28 and 42 dpi regarding the cerebellum. Earlier and more severe neurodegeneration during T. canis- than T. cati-induced NT, especially in the cerebrum, may explain the differences in behavioural alterations observed in previous studies. In addition to differences in larval migration preferences, immunological processes may contribute to these patterns, which warrant further investigation.

An innovative approach in the detection of Toxocara canis excretory/secretory antigens using specific nanobodies.

Human toxocariasis is a zoonosis resulting from the migration of larval stages of the dog parasite Toxocara canis into the human paratenic host. Despite its well-known limitations, serology remains the most important tool to diagnose the disease. Our objective was to employ camelid single domain antibody fragments also known as nanobodies (Nbs) for a specific and sensitive detection of Toxocara canis excretory/secretory (TES) antigens. From an alpaca immune Nb library, we retrieved different Nbs with specificity for TES antigens. Based on ELISA experiments, these Nbs did not show any cross-reactivity with Ascaris lumbricoides, Ascaris suum, Pseudoterranova decipiens, Anisakis simplex and Angiostrongylus cantonensis larval antigens. Western blot and immunocapturing revealed that Nbs 1TCE39, 1TCE52 and 2TCE49 recognise shared epitopes on different components of TES antigen. The presence of disulphide bonds in the target antigen seems to be essential for recognition of the epitopes by these three Nbs. Three separate sandwich ELISA formats, using monovalent and bivalent Nbs, were assessed to maximise the detection of TES antigens in solution. The combination of biotinylated, bivalent Nb 2TCE49 on a streptavidin pre-coated plate to capture TES antigens, and Nb 1TCE39 chemically coupled to horseradish peroxidase for detection of the captured TES antigens, yielded the most sensitive ELISA with a limit of detection of 0.650 ng/ml of TES antigen, spiked in serum. Moreover, the assay was able to detect TES antigens in sera from mice, taken 3 days after the animals were experimentally infected with T. canis. The specific characteristics of Nbs make this ELISA not only a promising tool for the detection of TES antigens in clinical samples, but also for a detailed structural and functional study of TES antigens.

The serodiagnostic potential of recombinant proteins TES-30 and TES-120 in an indirect ELISA in the diagnosis of toxocariasis in cattle, horses, and sheep.

Toxocariasis is a zoonotic disease that affects humans and animals alike. Although recombinant proteins are widely used for its diagnosis in humans, their performance in companion and production animals remains unknown. This study aimed to investigate the serodiagnostic potential of the recombinant proteins rTES-30 and rTES-120 from Toxocara canis in an indirect ELISA for cattle, horses, and sheep. Serum samples collected from the animals were tested with indirect ELISA and Western Blotting using T. canis TES-30 and TES-120 recombinant proteins produced in Escherichia coli, as well as native-TES. In the ELISA, rTES-30 showed high serodiagnostic potential in sheep and horses (92.6% and 85.2%, respectively), while the sensitivity of rTES-120 was higher in cattle and horses (97.2% and 92.6%, respectively). Furthermore, a highly positive association was observed between native and recombinant proteins in seropositive samples, while a moderately positive association was observed in seronegative samples, probably due to the lower specificity of native TES. In conclusion, our study indicates that the use of recombinant proteins in an indirect ELISA is an effective tool for the serodiagnosis of toxocariasis in animals, with the choice of protein being species-dependent.

Human toxocariasis seroprevalence among patients with uveitis in Alborz Province, Iran.

INTRODUCTION AND OBJECTIVE: Toxocariasis, predominantly caused by Toxocara canis, is a common zoonotic parasitosis worldwide. Toxocara infection is a cause of vision impairment and blindness. The presented study investigates the frequency of antibodies against Toxocara among uveitis patients and the epidemiological factors associated with disease. MATERIAL AND METHODS: Fifty-four patients with uveitis and 59 healthy subjects were studied. Anti-Toxocara antibodies status was determined in all serum samples using enzyme linked immunosorbent assay (ELISA), and seropositive samples analyzed by Western blot (WB) technique. RESULTS: The frequency of Toxocara canis infection was found to be significantly higher in uveitis patients, compared to healthy controls by the use of ELISA test, being 14.8% and 1.7%, respectively. From 8 seropositive samples, 5 (62.5%) patients exhibited Toxocara immunoglobulin G (IgG) antibodies in response to Western blot, whereas in the control group, none were detected positive by Western blot. No significant difference was found between pet owners, nor between different places of residence. The seroprevalence to Toxocara among uveitis patients was significantly related to gender, age and medical diagnosis. The highest prevalence was found in patients with posterior uveitis (27.8%). CONCLUSIONS: Anti-Toxocara antibody titers are associated with the risk of vision impairment -uveitis. The risk factor associated with Toxocara exposure identified in this study warrants further investigation.

Diagnostic Potential of Anti-RTE30 Polyclonal Antibodies In A Blocking Elisa For Toxocara canis Detection.

The main etiologic agent of human toxocariasis, a zoonotic disease, is the helminth Toxocara canis. Among the diagnostics used for human toxocariasis, ELISA using T. canis excretion and secretion antigen (TES) is considered as a standard technique. TES antigen requires the cultivation of T. canis larvae, which makes its production difficult. Besides this, the use of TES antigen does not eliminate the cross-reactions with other similar proteins that are produced by other intestinal worms. In this context, recombinant antigens are being tested to improve the diagnosis of human toxocariasis. Herein, we describe the production of polyclonal antibodies against recombinant protein TES30 (pAb-rTES30) and evaluate its use in a blocking ELISA (b-ELISA) using human sera. The b-ELISA showed 95.6% sensitivity and 94.4% specificity. Thus, the b-ELISA using pAb-rTES30 offers a viable option for toxocariasis diagnosis owing to its configuration, which prevents cross-reactivity with non-species-specific antibodies.

Matrix metalloproteinase-2 and matrix metalloproteinase-9 in mice with ocular toxocariasis.

In ocular toxocariasis, Toxocara canis-induced inflammatory reaction can lead to eye destruction and granuloma, which is formed by immune cell infiltration and concurrent extensive remodeling tissue. Herein, the histomorphology of granuloma and proteinase production in the eye of T. canis-infected BALB/c mice were investigated. Pathological effects substantially increased after the infection culminated in a severe leukocyte infiltration and granuloma formation from days 4 to 56 post-inoculation. The matrix metalloproteinase (MMP)-2 and MMP-9 activities remarkably increased, compared with those of uninfected control, by gelatin zymography and Western blot analysis in ocular toxocariasis. Granuloma formation had a remarkably positive correlation with MMP-2 and MMP-9 levels. We suggested that T. canis larvae and leukocytes infiltrated from blood vessel both migrated into corpus adiposum orbitae. Activated leukocytes secreted MMP-2 and MMP-9, leading to fibronectin degradation. The imbalance of MMP-2/TIMP-2 and MMP-9/TIMP-1 may play a role in inflammatory cell infiltration and extracellular matrix degradation, forming granuloma, in ophthalmological pathogenesis of T. canis infection.

Space Occupying Lesion in the Liver Caused by Hepatic Visceral Larva Migrans: A Case Report.

Visceral larva migrans (VLM) is one of the clinical syndromes of human toxocariasis. We report a case of hepatic VLM presenting preprandial malaise and epigastric discomfort in a 58-year-old woman drinking raw roe deer blood. The imaging studies of the abdomen showed a 74-mm hepatic mass featuring hepatic VLM. Anti-Toxocara canis immunoglobulin G (IgG) was observed in enzyme-linked immunosorbent assay (ELISA) and western blot. Despite anthelmintic treatment, the patient complained of newly developed cough and skin rash with severe eosinophilia. Hepatic lesion increased in size. The patient underwent an open left lobectomy of the liver. After the surgery, the patient was free of symptoms such as preprandial malaise, epigastric discomfort, cough, and skin rash. Laboratory test showed a normal eosinophilic count at postoperative 1 month, 6 months, 1 year, and 4 years. The initial optical density value of 2.55 of anti-T. canis IgG in ELISA was found to be negative (0.684) at postoperative 21 months. Our case report highlights that a high degree of clinical suspicion for hepatic VLM should be considered in a patient with a history of ingestion of raw food in the past, presenting severe eosinophilia and a variety of symptoms which reflect high worm burdens. Symptom remission, eosinophilia remission, and complete radiological resolution of lesions can be complete with surgery.

Seroprevalence of ToxocaraCanis in Asthmatic Children and its Relation to the Severity of Diseases - a Case-Control Study.

Background: Toxocariasis is a common parasitic infectionworldwide even in developed countries. Through this health problem, the immune system is triggered and the antibody is produced, leading to some hypersensitive situations like asthma. In the present study, we tried to show a correlation between Toxocara Canis (T. canis) seropositivityand asthma in Isfahan city. Methods: This research is a cross-sectional study involving 40 asthmatics and 46 non-asthmatic cases aged 2-18 years. In all cases, T. canis IgG level was measured using enzyme-linked immunosorbent assay (ELISA) and compared between two groups. Results: The seroprevalence of IgG anti-T. canis antibodies were 45%in the asthmatic patients and 21.7% in thecontrols (P= 0.022). The more severe asthmatic patients had significantly more risk for T. canis seropositivity (P= 0.019). In the asthmatic patients, there was a significant correlation between Toxocara seropositivity and goingto park or playground (P= 0.001). Conclusion: In this study, we found a significantly positive serology of T. canis in asthmatic children especially in more severe disease. To verify the etiologic role of Toxocara in asthma, more advanced studies are needed.

Detection of Toxocara canis DNA in tissues of experimentally infected mice.

The main etiological agent of toxocariasis is the helminth Toxocara canis. Several difficulties are found in the diagnosis of this disease, because of nonspecific clinical signs and possible cross-reactions that may occur in the available test, the indirect ELISA. Therefore, molecular diagnosis has been indicated as an alternative to conventional diagnosis. The purpose of this study was to evaluate the polymerase chain reaction (PCR) technique for the identification of T. canis in tissues of experimentally infected mice. To this end, nine mice were inoculated with 1500 embryonated eggs and were divided into two groups, the first euthanized 48 h (G1) and the other 30 days post inoculation (G2). Lungs, brain, liver and blood were collected from all the animals for DNA Extraction and tissue digestion, also was collected blood samples for DNA extraction and ELISA test (serum). Toxocara canis DNA was identified in all the inoculated animals using the ITS-2 target gene. The PCR test successfully identified the parasite in the brain, lung and liver of the animals euthanized 48 h PI and 30 days PI. This technique yielded good results in the identification of the parasite in the brain, being more sensitive than the method for the recovery of larvae, in the group with acute infection (48 h PI). The infection was confirmed by PCR within 48 h after infection, while the ELISA indicated serological conversion occurred only 14 days after inoculation. This study demonstrates the ability of PCR to identify T. canis in the liver, lungs and brain during acute and chronic infection.

Seroprevalence of Toxoplasma gondii and Toxocara canis in a human rural population of Southern Rio Grande do Sul.

Due to the growing population of pets, especially homeless dogs and cats, zoonoses still represent a significant public health problem. Toxoplasma gondii and Toxocara spp. are epidemiologically important zoonotic agents as they are etiological factors of human toxoplasmosis and toxocariasis, respectively. These parasites remain neglected even though they are substantially prevalent in rural areas. The aim of this study was to investigate T. gondii and T. canis seroprevalence and risk factors of seropositivity in a rural population in Pelotas municipality, Brazil. The study participants (n=344) were patients of a Basic Healthcare Unit (BHU) located in Cerrito Alegre. Blood samples were collected and tested for T. gondii antibodies by indirect immunofluorescence and T. canis antibodies by an indirect ELISA that targets an excreted-secreted antigen (TES). T. gondii seropositivity was 53.2%, with higher titers (1:256 - 1:1,024) in individuals who habitually eat pork, beef, or chicken, while T. canis seropositivity was 71.8% and concomitant T. gondii and T. canis seropositivity was 38.3%. Among the seropositivity risk factors assessed, only habitual undercooked meat consumption was significant (p = 0.046; OR = 3.7) for T. gondii and none of them were associated with T. canis seropositivity. Both parasites have a high prevalence in rural areas, which reinforces the need to invest in rural community education and health.

Reactivity of recombinant Toxocara canis TES-30/120 in experimentally infected mice.

AIM: While the use of recombinant antigens is being widely investigated in the diagnosis of human toxocariasis, relatively little attention has been given to animal diagnostic models. For this reason, this study aimed to investigate the diagnosis potential of Toxocara canis TES-30 and TES-120 recombinant antigens in mice, the animal model for toxocariasis studies. METHODS AND RESULTS: Serum samples obtained from mice infected with T. canis or Toxocara cati were tested by indirect ELISA using T. canis TES-30 and TES-120 recombinant antigens produced in Escherichia coli. 90% of the samples reacted with rTES-30, whereas there was almost no reactivity with rTES-120. CONCLUSION: Despite rTES-120 being a good antigen for diagnosis in humans, it could not reproduce its reactivity in this animal model. As rTES-30 has good reactivity in mice, it is a valuable tool for diagnosis.

Prolactin as immune cell regulator in Toxocara canis somatic larvae chronic infection.

Toxocariasis is a zoonotic disease produced by ingestion of larval Toxocara spp. eggs. Prolactin (PRL) has been considered to have an important role in Toxocara canis infection. Recent evidence has found that PRL directly can increase parasite growth and differentiation of T. canis The present study, evaluated the effect of high PRL levels on the immune system's response and parasites clearance in chronic infection. Our results showed that hyperprolactinemia did not affect the number of larvae recovered from several tissues in rats. Parasite-specific antibody production, showed no difference between the groups. Lung tissue presented eosinophilic granulomas typical of a chronic infection in all the experimental groups. Flow cytometry analysis was made in order to determine changes in the percentage of innate and adaptive immune cell subpopulations in the spleen, peripheric (PLN) and mesenteric (MLN) lymphatic nodes. The results showed a differential effect of PRL and infection on different immune compartments in the percent of total T cells, T helper cells, T cytotoxic cells, B cells, NK cells, and Tγδ cells. To our knowledge, for the first time it is demonstrated that PRL can have an immunomodulatory role during T. canis chronic infection in the murine host.