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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.)

Probiotics have been shown to reduce the intensity of Toxocara canis infection in mice. However, larval transmission of this nematode also occurs via transplacental and transmammary routes. Thus, the aim of this study was to evaluate the effect of the Saccharomyces boulardii probiotic on the vertical transmission of T. canis in Swiss mice. The mice received 107S. boulardii colony-forming units per gram of food. The supplementation began 15 days before mating and was maintained throughout pregnancy and lactation. The animals were inoculated with 300 T. canis embryonated eggs on the 14th day of pregnancy. The presence of larvae was examined in the organs of the females and their offspring. The examined organs included the following: brain, liver, lungs, heart, kidneys, spleen, eye, skeletal muscle (carcass) and mammary glands of lactating females. There was a 42% (P = 0.041) reduction in the number of larvae transmitted to offspring in the group that received probiotic-supplemented food (GI). Additionally, there was a 50% reduction (P = 0.023) in the number of larvae found in the brains of lactating offspring in the GI group. These results reveal the potential of S. boulardii probiotic use as an auxiliary method of controlling visceral toxocariasis.

Human toxocariasis consists of chronic tissue parasitosis that is difficult to treat and control. This study aimed to evaluate the action of the probiotic Lactobacillus acidophilus ATCC 4356 on larvae of Toxocara canis and the effect of IFN-γ cytokine on parasite-host in vivo (1.109 CFU) and in vitro (1.106, 1.107, 1.108, 1.109 CFU) interactions. Four groups of six BALB/c mice were formed: G1 - L. acidophilus supplementation and T. canis infection; G2 - T. canis infection; G3 - L. acidophilus supplementation; and G4 - PBS administration. Mice were intragastrically suplemented with probiotics for 15 days before inoculation and 48 h after inoculation with 100 T. canis eggs. The inoculation of T. canis was also perfomed intragastrically. The recovery of larvae took place through digestion of liver and lung tissues; the evaluation of IFN-γ gene transcription in leukocytes was performed by qPCR. The in vitro test consisted of incubating the probiotic with T. canis larvae. The supplementation of probiotics produced a reduction of 57.7% (p = 0.025) in the intensity of infection of T. canis larvae in mice, whereas in the in vitro test, there was no larvicidal effect. In addition, a decrease in the IFN-γ gene transcription was observed in both, T. canis-infected and uninfected mice, regardless of whether or not they received supplementation. The probiotic L. acidophilus ATCC 4356 reduced T. canis infection intensity in mice, however, the probiotic did not have a direct effect on larvae, demonstrating the need of interaction with the host for the beneficial effect of the probiotic to occur. Yet, the proinflammatory cytokine IFN-γ did not apparently contributed to the observed beneficial effect of probiotics.

The nematode Toxocara canis is of public health importance and is the main causative agent of toxocariasis in humans. This disease is difficult to diagnose due to several factors, including the possibility of cross-reactions with other nematodes in the ELISA. To overcome this problem, molecular tests have been recommended as an alternative to identify the parasite. The quantitative real-time polymerase chain reaction (qPCR) technique was used in this study to identify and quantify the parasite load of T. canis in the mouse brain. To this end, 24 mice were divided into six groups, five of which were challenged with different infective doses of T. canis larvae (L3) (1000, 500, 250, 100 and 50 larvae), while the sixth group, uninfected, acted as negative control. Forty-five days after infection, the animals were euthanized to collect the brain, from which two portions of 20 mg of tissue were taken for DNA extraction, while the rest of the brain tissue was digested to quantify the number of larvae by microscopy. The number of DNA copies was calculated from the standard DNA quantification curve, showing values of E = 93.4%, R2 = 0.9655 and Y = -3.415. A strong positive correlation (R = 0, 81; p < .001) was found between the number of copies and the recovery of larvae from brain. However, the parasite's DNA was also identified even in animals from whose brain no larvae were recovered after tissue digestion. The results of this study therefore confirm that the qPCR technique can be a valuable tool for the detection and quantification of T. canis DNA in murine hosts, even in animals whose with tissues contain very few parasites., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Introduction: Day care centers play an important social role in children's early education and development. The objective of this study was to investigate the frequency of intestinal parasitic infections and infection with Toxocara spp. in children from a day care center in the city of Rio Grande, Rio Grande do Sul, Brazil. Methods: The study was conducted using fecal samples from 50 children, ages three to six years, using Ritchie, Faust, Rugai, and Kinyoun's techniques. A closed and structured epidemiological questionnaire was used to collect data from the parents/guardians of the children. Serological studies for detection of antibodies to Toxocara spp. by immunoenzymatic assays using Toxocara excretion and secretion antigen (TES) were conducted on sera collected from 41 children. Results: The frequency of enteroparasites was 18%, with 43.9% of the children testing seropositive for Toxocara spp. Low family income and low literacy levels of parents/guardians were common factors between the families of the parasite-positive children. For the children who tested positive for Toxocara spp., most of the parents/guardians reported using only water for sanitizing raw vegetables and fruits. Conclusions: The high seropositivity rates for Toxocara spp. indicate that children were exposed to this parasite, and it is important to reduce the risk of infection. In addition, the seropositivity for enteroparasites and the interviews with the parents/guardians indicate the need to educate the studied population regarding the modes of intestinal parasite transmission and the prophylactic measures needed to prevent their dissemination.

Probiotics have shown promising results as a potential method to control toxocariasis in mice inoculated with embryonated eggs of Toxocara canis. This study aimed to evaluate the protective effect of Saccharomyces boulardii in mice fed in natura chicken livers infected with T. canis. Twenty 15-day-old male Sussex chickens were inoculated with 300 T. canis embryonated eggs via intragastric catheter (GI). After 72 h of infection, each liver was collected and individually offered to a group of 20 mice. Mice that received supplemented ration with S. boulardii (1.10 7 colony forming units) and consumed in natura chicken liver showed reduction in infection intensity of 67.1%. This study demonstrated that administration of S. boulardii has potential as a probiotic to assist in controlling visceral toxocariasis caused by the consumption of viscera from paratenic hosts containing infective parasite larvae.

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., Competing Interests: The authors have declared that no competing interests exist.

Toxocariasis is a neglected zoonosis that affects children and adults. Recombinant proteins have been widely investigated for diagnosis, achieving high sensitivity and specificity in an overall population; however, little is known about age as a factor in its application. This study aims to investigate the diagnostic potential of Toxocara canis TES-30 and TES-120 recombinant proteins in humans, differentiating between its performance in children and adults. Serum samples collected from children and adults seropositive to Toxocara spp. were tested with indirect ELISA using T. canis TES-30 and TES-120 recombinant proteins produced in Escherichia coli. While rTES-30 sensitivity was not affected by age (81.8% in children and 87% in adults), rTES-120 sensitivity severely decreased in children to only 63.6%, down from 95.7% in adults. Furthermore, the sensitivity of rTES-30 increased to 97.8% after Western blotting confirmation. High specificity (>94%) against other geohelminths was reported for both recombinant proteins. Our study favors the use of rTES-30 with total IgG as the primary antibody in an indirect ELISA assay as a tool for epidemiological human studies., Competing Interests: The authors have declared that no competing interests exist.

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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

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., (© 2018 John Wiley & Sons Ltd.)

Human toxocariasis is a neglected global parasitic zoonosis. The efficacy of drug treatment for this disease has been hindered by the biological complexity of the main etiological agent, the nematode Toxocara canis. Experimental studies have shown the potential of probiotics to promote a reduction in the parasite load of T. canis larvae. This study aimed to evaluate the effect of probiotic Lactobacillus rhamnosus ATCC 7469 on the parasite load of BALB/c mice with acute toxocariasis and evaluate the direct effect of this probiotic on T. canis larvae in vitro. In vivo administration of probiotics reduced the parasite load of T. canis larvae by 53.3% (p = 0.0018) during the early stage of infection in mice. However, when analyzed in vitro, it was observed that the probiotic did not present a deleterious effect on the larvae, as approximately 90% of these remained viable. These results demonstrate the potential of the probiotic L. rhamnosus in the reduction of T. canis larvae in BALB/c mice and suggest it could be used as an alternative means for the controlling of visceral toxocariasis. However, further studies are required to elucidate the mechanisms of action promoted by this probiotic.

Toxoplasmosis causes complications during pregnancy that have serious effects on fetal development. Thus far, toxocariasis has been reported to spread only via vertical transmission. Nonetheless, the population of pregnant women is also exposed to this infection. Co-infection with both Toxoplasma gondii and Toxocara spp. has been reported in children, but there are no reports of co-infection in the population of pregnant women. The aim of this study was to determine the prevalence of co-infection with T. gondii and Toxocara spp. in pregnant women at a university hospital in southern Brazil, and to identify the risk factors associated with infection by both parasites. Two hundred pregnant women were tested for the presence of anti-T. gondii and anti-Toxocara spp. antibodies and were asked to complete an epidemiological questionnaire. In this study, the co-infection rate observed in the total population of pregnant women was 8%. In addition, women with a positive result for a serology test for Toxocara spp. were at increased risk of infection by T. gondii (P = 0.019). Co-infection with both parasites in pregnant women was associated with low birth weights in neonates. The similar modes of transmission of both parasites could explain the co-infection. Only a few previous studies have investigated this phenomenon. The findings of the present study emphasize the importance of serological diagnosis during prenatal care and further research in this area to identify risk factors associated with this co-infection, and the possible implications of this co-infection during pregnancy and on the health of newborns.

Toxocariasis is a neglected and geographically widespread parasitic disease. The detection of specific antibodies associated with this disease is required to confirm its clinical diagnosis and to aid in prevention. Although helminth infection during pregnancy can promote foetal immune responses with long-term effects, specific information regarding the risk of Toxocara spp. infection to the human foetus during pregnancy is lacking. Therefore, the present study aimed to investigate the frequency of antibodies against Toxocara spp. in umbilical cord serum samples to determine the neonatal risk factors associated with Toxocara spp. infection. A cross-sectional study of the frequency of specific antibodies against Toxocara spp. was performed on umbilical cord samples of 280 neonates. A cord blood sample was obtained from each newborn after parturition, and serum samples were examined by enzyme-linked immunoassay. Epidemiological data were obtained through a questionnaire regarding obstetric history (abortion history, premature birthhistory, and pregnancy and birth numbers), general aspects (animal contact anddiet) and socio-economic factors. The frequency of anti-Toxocara spp. IgG antibodies in the umbilical cords of neonates was 20% in serum pre-adsorbed with Ascaris spp. antigen. Family income and dog ownership were considered risk factors associated with infection. No association was found between reproductive disorders and Toxocara seropositivity. The 20% frequency rate of anti-Toxocara spp. IgG antibodies in sera from umbilical cords of newborns can be related to IgG binding at the maternal-foetal interface, requiring greater care during pregnancy. Anti-Toxocara IgM and IgE antibodies no were found in umbilical cord serum samples, indicating that no vertical transmission of these parasites occurred in this population. Studies regarding antibodies against Toxocara spp. in umbilical cord sera are important for determining neonatal exposure to these parasites., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Trichomoniasis is the most prevalent non-viral sexually transmitted disease (STD) in the world; however, it remains a neglected parasitic disease. This study aimed to determine the prevalence of trichomoniasis and its associated epidemiological factors among women treated at a hospital in southern Brazil., Methodology/principal Findings: A cross-sectional study was performed to determine the prevalence of this infection in women treated at Hospital Universitário (HU) in Rio Grande, Rio Grande do Sul, Brazil, between January 2012 and January 2015. This study consisted a self-administered questionnaire regarding demographic, clinical, and behavioural data and a molecular diagnosis with polymerase chain reaction (PCR) using the TVK3/7 primer set, which was confirmed with sequence analysis. Of the 345 women surveyed, the overall prevalence of Trichomonas vaginalis (T. vaginalis) was 4.1% (14/345). The prevalence rates were 5.9% among pregnant women, 8.5% among HIV-positive women, and 10.1% among HIV-positive pregnant women. The rates for groups with other significant demographic and clinical features were as follows: 6.6% among women with white skin, 12.3% among women with an income below the minimum monthly wage, 7.4% among women with a vaginal pH greater than or equal to 4.6, and 7.9% among women with a comorbid STD. The multivariate analysis confirmed that pregnant women who were HIV-positive (p = 0.001) and had low incomes (p = 0.026) were the most likely to have this infection., Conclusions: A multivariate analysis confirmed that HIV-positive pregnant women with low incomes were the participants most likely to have trichomoniasis. These results are important because this Brazilian region presents a high prevalence of HIV-1 subtype C, which is associated with greater transmissibility. Additionally, low family income reveals a socioeconomic fragility that might favour the transmission of this STD.

Human toxocariasis is a neglected parasitic disease worldwide. Researchers studying this disease use infectious strains of Toxocara for experiments. Health workers are at risk in the course of their daily routine and must adhere to biosafety standards while carrying out the activities. Researchers on biosafety concerning working with these parasites are insufficient. The aim of this study was to determine the rate of seroprevalence of Toxocara species among health-care research laboratory workers (professors, technicians, and students), and to investigate the risk factors of Toxocara infection associated with laboratory practices. This cross-sectional study involved 74 researchers at two federal universities in southern Brazil from February 2014 to February 2015; 29 researchers manipulated infective strains of Toxocara canis (test group) and 45 did not (control group). Serum samples were examined using enzyme-linked immunosorbent assay. Epidemiological data were obtained via a questionnaire containing information about laboratory routine, eating behavior, and contact with dogs. The seroprevalence of anti-T. canis IgG was 14.9% (11/74; 13.8% [4/29] in the test group and 15.6% [7/45] in the control group). Most individuals in the test group correctly understood the primary mode of infection; however, 13.8% did not use gloves while manipulating T. canis eggs. Knowledge of biosafety must be well understood by health-care professionals doing laboratory work with biological agents. To our knowledge, this is the first study to investigate the rate of seroprevalence of IgG against Toxocara spp. among professionals and students who handle infective forms of the nematode T. canis., (© The American Society of Tropical Medicine and Hygiene.)

Introduction  Toxoplasmosis a parasitic zoonosis of global distribution, responsible for disorders during gestation can cause fetal death or congenital anomalies. Objective  To evaluate the knowledge of toxoplasmosis among pregnant and postpartum women treated at the University Hospital of the city of Rio Grande, Rio Grande do Sul, Brazil. Methods  This was a cross-sectional study of 100 pregnant and postpartum women at the University Hospital. Participants answered a self-administered questionnaire and gave consent for data relating to serological examinations to be abstracted from their medical records. Results  The proportion of women who received information about toxoplasmosis was higher among those who received care in the private health care system (52.9%) than among those cared for in the public health care system (25.0%). Only 55.7% of women reported having some knowledge about toxoplasmosis. Of these, 53.7% received information during the prenatal period. However, most participants were unable to answer questions about preventive measures and modes of infection. Of the 100 patients in the study, only 46 underwent serologic testing for toxoplasmosis, 65.2% of whom tested negative (IgG). Conclusion  Findings from this study are relevant to the training of health professionals regarding toxoplasmosis education and prevention. Improved education for health care providers and patients can lead to earlier diagnoses and reductions in adverse outcomes., (Thieme Publicações Ltda Rio de Janeiro, Brazil.)

Objective: The objective of this study is to investigate the association between anti-Toxocara IgG seropositivity and asthma in children., Methods: This was a case-control study conducted in a university hospital in south Brazil between May 2012 and June 2013. Were recruited 208 children up to 12 years old of whom 156 had asthma (cases) and 52 did not have asthma (controls), with a case-control ratio of 3:1 matched by age. Children's parents or guardians were interviewed using a structured questionnaire with closed questions. Serology was performed using enzyme-linked immunosorbent assay (ELISA) with excretory-secretory antigen of Toxocara canis (TES)., Results: The seroprevalence of IgG anti-T. canis antibodies was 12.8% in the cases and 7.7% in the controls. There was no significant association between seropositivity to T. canis and risk of asthma (adjusted odds ratio [OR]: 1.89, 95% CI: 0.52 to 6.89, p = 0.33). Household income < 2 minimum salaries, paternal school years < 9, allergic rhinitis in children, a positive family history of asthma and rhinitis and contact with cats were significantly associated with asthma, with adjusted ORs (95% CIs) of 3.05 (1.21 to 7.73), 2.83 (1.11 to 7.18), 10.5 (4.32 to 25.6), 2.65 (1.14 to 6.17), 2.49 (1.07 to 5.78) and 2.73 (1.03 to 7.27), respectively., Conclusions: This study did not find a statistically significant association between seropositivity to Toxocara sp. and risk of asthma in children. Low family income, low paternal education level, concomitant allergic rhinitis, family history of asthma and allergic rhinitis and contact with cats were independent factors associated with childhood asthma.

In this study, supplementation with the probiotic Saccharomyces boulardii promoted a reduction in intensity of infection by Toxocara canis and modulates cytokines mRNA expression in experimentally infected mice. IL-12 gene transcription had 40-fold increase in S. boulardii supplemented uninfected mice and sevenfold increase in supplemented infected mice comparing with not supplemented group. Regarding IFNγ, similar results were observed, since probiotic supplementation induced approximately 43-fold increase, but only in uninfected mice (P < 0·05). T. canis infection upregulated IL-10 expression while S. boulardii downregulated it and no change was observed for IL-4. Thus, based in these findings; we suggest that one possible mechanism responsible for S. boulardii protection effect against T. canis infection is by the modulation of cytokines expression, especially IL-12., (© 2016 John Wiley & Sons Ltd.)

Human toxocarosis is a chronic tissue parasitosis most often caused by Toxocara canis. The seroprevalence can reach up to 50%, especially among children and adolescents. The anthelmintics used in the treatment have moderate efficacy. The aim of this study was to evaluate the in vitro and in vivo anthelmintic activity of quinones and their derivatives against T. canis larvae and the cytotoxicity of the larvicidal compounds. The compounds were evaluated at 1 mg mL(-1) concentration in microculture plates containing third stage larvae in an Roswell Park Memorial Institute (RPMI) 1640 environment, incubated at 37 °C in 5% CO2 tension for 48 h. Five naphthoxiranes were selected for the cytotoxicity analysis. The cell viability evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assays using murine peritoneal macrophages isolated from C57BL/6 mice revealed that the naphthoxiranes (1 and 3) were less cytotoxic at a concentration of 0.05 mg mL(-1). The efficacy of naphthoxiranes (1 and 3) was examined in murine toxocarosis also. The anthelmintic activity was examined by evaluating the number of larvae in the brain, carcass, liver, lungs, heart, kidneys and eyes. Compound (3) demonstrated anthelmintic activity similar to that of albendazole by decreasing the number of larvae in the organs of mice and thus could form the basis of the development of a new anthelmintic drug.

Considering the therapeutic potential of fatty acid amides, the present study aimed to evaluate their in vitro activity against Toxocara canis larvae and their cytotoxicity for the first time. Linoleylpyrrolidilamide was the most potent, with a minimal larvicidal concentration (MLC) of 0.05 mg/mL and 27% cytotoxicity against murine peritoneal macrophages C57BL/6 mice, as assessed by the MTT assay., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: Human toxocariasis is a parasitic zoonosis with a worldwide distribution but is underdiagnosed with an underestimated impact on human health. The ingestion of embryonated eggs of Toxocara spp. present on the hands or in contaminated food or water is the main mode of infection. The only record of Toxocara congenital infection in humans occurred in a premature infant. Helminth infections during pregnancy may be associated with reproductive disorders. Studies investigating the occurrence of toxocariasis in pregnancy are scarce, as is research on the possible implications of these parasites in reproductive health. The aim of this study was to determine the seroprevalence of antibodies to Toxocara spp. in pregnant women and to identify risk factors associated with its infection., Methodology/principal Findings: The cross-sectional study of the seropositivity of specific antibodies for Toxocara spp. was performed on 280 pregnant women. Serum samples were examined with enzyme-linked immunoassay. Epidemiological data were obtained through a questionnaire containing information about obstetric history, general life style choices, and the social and economic status of the women. The prevalence of Toxocara spp. IgG in pregnant women was 6.4%. Some of the risk factors associated with the infection were owning dogs (p = 0.003), living in the city centre (p = 0.028), living at the city beach (p = 0.003), and having a family income at or below minimum wage (p < 0.001). There was no association between reproductive disorders and Toxocara seropositivity., Conclusions/significance: The seroprevalence of 6.4% for Toxocara spp. in pregnant women shows that there was exposure to the parasite. The study demonstrates the need for attention for the completion of clinical diagnosis parameters, as well as the expansion of highly specific serological studies in different regions to understand the impact of toxocariasis in pregnancy.

Anthelmintics used for intestinal helminthiasis treatment are generally effective; however, their effectiveness in tissue parasitosis (i.e. visceral toxocariasis) is moderate. The aim of this study was to evaluate the in vitro activity of lapachol, β-lapachone and phenazines in relation to the viability of Toxocara canis larvae. A concentration of 2 mg/mL (in duplicate) of the compounds was tested using microculture plates containing Toxocara canis larvae in an RPMI-1640 environment, incubated at 37 °C in 5% CO2 tension for 48 hours. In the 2 mg/mL concentration, four phenazines, lapachol and three of its derivatives presented a larvicide/larvistatic activity of 100%. Then, the minimum larvicide/larvistatic concentration (MLC) test was conducted. The compounds that presented the best results were nor-lapachol (MLC, 1 mg/mL), lapachol (MLC 0.5 mg/mL), β-lapachone, and β-C-allyl-lawsone (MLC, 0.25 mg/mL). The larvae exposed to the compounds, at best MLC with 100% in vitro activity larvicide, were inoculated into healthy BALB/c mice and were not capable of causing infection, confirming the larvicide potential in vitro of these compounds.

Human toxocariasis is a neglected public health problem. Infection of humans generally results from the accidental ingestion of embryonated Toxocara canis eggs, but it is important to broaden knowledge about other forms of transmission. This study aimed to demonstrate the prevalence of transmammary transmission in mice with chronic toxocariasis. BALB/c mice in groups 1 (G1) and 3 (G3) were inoculated with 1200 T. canis eggs 60days before mating, whereas those of group 2 (G2) were not infected. After delivery, the G1 neonates were transferred to G2 females to be nursed, and vice versa. Thus, the mice generated by G2 females and breastfed by G1 females could be infected only during lactation. In the G3 group, offspring were not exchanged. The search for T. canis larvae in the bodies of the lactating females and their offspring was performed after weaning and at 60days old, respectively. The frequency of transmammary infection in the mice generated by G2 uninfected females and breastfed by G1 infected females was 19.8%, which was similar to that observed (19.6%) in the mice bred and fed by G3 females. The frequency of infection in the mice generated by G1 females and breastfed by G2 females was only 4.2%, which was lower than that of G1 (p=0.0064) and G3 (p=0.0062) groups. Transmammary infection by mice with chronic toxocariasis was found to be more prevalent than congenital infection., (Copyright © 2014. Published by Elsevier Ireland Ltd.)

Toxocariasis is a widespread zoonosis and is considered an important worldwide public health problem. The aim of this study was to investigate the frequency of trans-mammary Toxocara canis infection in newborn BALB/c mice nursed by females experimentally infected with 1,200 eggs after delivery. After 50 days of age, the presence of larvae in different organs of the offspring was investigated. Trans-mammary infection was confirmed in 73.9% of the mice that had been nursed by infected females. These data show a high trans-mammary transmission of T. canis and confirm the significance of this transmission route in paratenic hosts.

Toxocariasis is a neglected zoonosis caused by the nematodes Toxocara canis and Toxocara cati. This disease is widespread in many countries, reaching high prevalence independently of the economic conditions. However, the true number of cases of toxocariasis is likely to be underestimated owing to the lack of adequate surveillance programs. Although some diagnostic tests are available, their sensitivity and specificity need to be improved. In addition, treatment options for toxocariasis are limited and are non-specific. Toxocariasis is listed as one of the five most important neglected diseases by the CDC. This review presents recent advances related to the control of toxocariasis, including new immunodiagnostics, therapies, and drug formulations, as well as novel interventions using DNA vaccines, immunomodulators, and probiotics., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Toxocariasis is a zoonotic disease in that IgM titers can remain high for long periods making difficult to determine the stage of the disease. The aim of this study is to investigate the applicability of indirect ELISA, associated with urea, to discriminate between the acute and chronic toxocariasis. IgG avidity was evaluated in 25 BALB/c mice experimentally infected with 1000 Toxocara canis eggs. Blood samples were collected, and sera treated with 6 M urea and assayed by ELISA every two weeks. The percent IgG avidity was determined using the mean absorbance of sera treated with urea, divided by the mean absorbance of untreated sera. In the first 15 days post-inoculation, was observed a low percentage, between 7.25 and 27.5%, IgG avidity, characteristic of an acute infection. After 60 days of infection, all the mice showed between 31.4 and 58% IgG avidity, indicating a chronic infection.

Experimental studies and registries of cases of human toxocariasis have shown that the consumption of raw or undercooked offal of the paratenic host of Toxocara canis may pose a risk of infection. Thus, we evaluated the risk of infection due to the consumption of liver of chickens inoculated with different doses of embryonated T. canis eggs. Doses were 5-100 times smaller than the ones previously employed in this type of study. Groups of five chickens were inoculated with 5000 (control), 1000, 500, 300 or 50 eggs of T. canis, and at 72 h post-inoculation, the liver of each bird was consumed by a BALB/c receptor mouse. Forty-eight hours after consumption, we examined the organs and carcasses of the mice for larvae of T. canis. All mice were positive for larvae, except the group that consumed the chicken liver inoculated with 50 eggs. This group contained only one positive mouse, in which the larva was lodged in the brain. In mice that consumed livers of chickens inoculated with ≥300 eggs, larvae concentration was primarily in the liver and lungs, characterizing the initial phase of infection. We conclude that the consumption of raw poultry liver, under the studied conditions, poses a risk of infection even with a low number of infected T. canis eggs., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Visceral toxocariasis is a neglected zoonosis caused by Toxocara canis larvae in unusual hosts. In dogs, the definitive host, the infection occurs mainly through transplacental and transcolostral transmission. Studies on experimental models have shown that vertical transmission may result from acute infections. Considering that toxocariasis is characterized as a chronic infection, with possible reactivation of larvae present in the brain, this study evaluated the presence of larvae in the brain of female BALB/c mice and their offspring with chronic infection during three successive pregnancies. ELISA-TES was used to evaluate the antibody levels. T. canis larvae were detected in the brain tissue of the mice during the three successive generations evaluated. The offspring's IgG level gradually decreased, and mean absorbance (ABS) above the cutoff point (0.070) was observed only at 30 (0.229) and 50 (0.096) days of age, while IgM was not detected. The infections in the offspring confirmed that vertical transmission of T. canis larvae occurred during chronic toxocariasis in three successive generations of mice.

Visceral toxocariasis is a neglected parasitic zoonosis that occurs through the ingestion of embryonated Toxocara spp. eggs. A wide range of animal species can act as paratenic hosts for this ascarid. The main risk factor for humans is the ingestion of the eggs from contaminated soil; however, infection can also occur through the ingestion of contaminated raw or undercooked infected meat from paratenic hosts. The aim of this study was to verify the presence of Toxocara spp.-specific antibodies in sheep and to determine the risk factors associated with the infection of sheep in Rio Grande do Sul (a major sheep-producing and sheep-consuming state) in southern Brazil. Serum samples collected from 1,642 sheep were tested using an IgG enzyme-linked immunosorbent assay based on the excretory-secretory Toxocara canis antigen. Seroprevalence was 29.0% (477/1,642), and every farm included in the study contained at least one seropositive animal. These results indicate that T. canis infection is widely distributed among sheep herds in Rio Grande do Sul and that it represents a potential risk to human health.

In a previous study our group found that the probiotic Saccharomyces boulardii was capable of reducing the intensity of infection in mice with toxocariasis. In order to assess whether the mechanism involved would be a direct action of the probiotic on Toxocara canis larvae, this study was designed. Both probiotics were singly cultivated in plates containing RPMI 1640 medium and T. canis larvae. S. boulardii and B. cereus var. toyoi cultures presented 97.6% and 95.7% of larvae with positive motility, respectively, and absence of color by the dye trypan blue, not representing significant difference to the control group (p > 0.05). We conclude that none of the probiotics showed in vitro effects on T. canis larvae and that the interaction with the intestinal mucosa is necessary for the development of the protective effect of S. boulardii.

The seroprevalence of Toxocara canis antibodies in children aged from 1 to 12 yr old was evaluated in Pelotas City, Rio Grande do Sul, Brazil. Human toxocariasis or visceral larva migrans (VLM) was diagnosed with the use of an ELISA based on the T. canis excretory-secretory (TES) antigens; Western blotting was used to confirm the ELISA-positive results. From 427 samples, 50.6% were positive for the presence of anti-TES antibodies. A confirmatory test (Western blot) was carried out on a sample of the ELISA-positive sera (n = 70), and all were positive. The Western blots had specific banding pattern characteristics, where the 30-kDa fraction demonstrated the highest reactivity. This fraction could be important for the specific diagnosis of toxocariasis.

Human toxocariasis is a neglected parasitic zoonosis of worldwide distribution. The consumption of raw or undercooked meat and offal from paratenic hosts of the Toxocara canis nematode can cause infection in humans, but there have been a lack of studies examining specific prophylactic measures to combat this mode of transmission. The aim of this study was to evaluate the establishment of infection by T. canis larvae at the initial and chronic phases of visceral toxocariasis after the consumption of mouse liver subjected to cold treatment. This study was divided into two stages using groups (G) of five donor mice inoculated with 2,000 eggs of T. canis. Two days post-inoculation, the livers of donor mice in G1 and G2 were kept at -20 °C and between 0 and 4 °C, respectively, for 10 days. In the first stage of the study, the livers of mice from G1, G2, and G3 (control) were subjected to a tissue digestion technique and found to be positive for infection. In the second stage, which evaluated infection in mice that had consumed livers from donor mice, receiver mice of G4 and G7 were fed with livers of donor mice from G1 (freezing), receiver mice of G5 and G8 were fed with livers of donor mice from G2 (cooling), and receiver mice of G6 and G9 with livers from G3 (control). Then, the tissue digestion technique was performed for recovering larvae from organs and carcasses of mice, at 2 days (G4, G5, and G6) and 60 days after liver consumption (G7, G8, and G9). It was observed that freezing inhibited the viability of 100 % of the larvae, while cooling promoted 87.7 and 95.7 % reductions in the intensity of infection at 2 and 60 days after liver consumption, respectively. Under the studied conditions, cold treatment shows great potential to help control this parasitosis, both in the initial and chronic phases of toxocariasis.

Helicobacter pylori infects nearly 50% of the world's population. This microorganism is accepted as the most important agent of gastritis and as a risk factor for peptic ulcer disease and gastric adenocarcinoma. Currently many diagnostic methods exist for detecting H. pylori, however they all have limitations, thus it is recommend a combination of at least two methods. The aim of this study was to evaluate diagnostic methods, such as in-house urease test, culture and Polymerase Chain Reaction (PCR), for the detection of the H. pylori in gastric biopsy specimens of 144 dyspeptic patients, using as gold standard the association between histology and rapid urease test. According to the gold standard used in this study, 48 (33.3%) patients were infected with H. pylori, while 96 (66.7%) were classified as not infected. The in-house urease test and the PCR were the most sensitive methods (100%), followed by culture (85.4%). However, the inhouse urease test and the culture were the most specific (100%), followed by PCR (75%). In conclusion, this study showed that, in comparison with the combination of histology and rapid urease test, the in-house urease test and the PCR presented 100% of sensitivity in the diagnosis of gastric infection by H. pylori, while the in-house urease test and the culture reached 100% of specificity. These finding suggest that the combination of two or more methods may improve the accuracy of the H. pylori detection.

Introduction: Tuberculosis (TB) control is linked to the availability of qualified methods for microbiological diagnostics; however, microscopy with limited sensitivity is the only method available in many locations. The objective of this study was to evaluate the introduction of culture, drug susceptibility testing (DST), and genotyping in the routine of a Municipal Program of Tuberculosis Control., Methods: Direct microscopy of sputum and culture in Ogawa-Kudoh were performed on 1,636 samples from 787 patients. DST of positive cultures was performed by resazurin microtiter assay and genotyping by mycobacterial interspersed repetitive units-variable number tandem repeat., Results: A total 91 patients with TB were identified. The culture increased case detection by 32% compared with the microscopy; acquired resistance was 3.3% and the genotyping showed high genetic diversity., Conclusions: Ogawa-Kudoh contributed significantly to the increase in case detection and is suitable for implementation in poor-resource locations. The acquired resistance rate was lower than that reported in a recent Brazilian survey. The high genetic diversity is possibly related to the high TB prevalence in the population, as well as to early detection and suitable treatment of patients. The interaction between research and health care is important for reorienting the practice, transferring technology, and improving TB control.

Several studies have shown the benefit of the use of probiotics in the prevention and treatment of diseases; however, few of them have investigated the effect of probiotics on parasitosis. In this study, the effect of Saccharomyces boulardii on the intensity of infection of mice with toxocariasis was evaluated. The animals were fed with a diet supplemented with S. boulardii for 15 days before inoculation with Toxocara canis eggs and for 2 or 60 days post-inoculation. S. boulardii promoted a reduction of approximately 36% in the average number of recovered T. canis larvae, suggesting that it can be used as an alternative to help control toxocariasis., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Visceral toxocariasis is a serious public health problem with a cosmopolitan distribution. Children are susceptible due to their immature immune system and high risks of infection. Nevertheless, the few completed studies about immunosuppression have had controversial results. To evaluate the effect of two immunosuppressive drugs on the larval burden of Toxocara canis, four groups of ten Swiss strain mice each were inoculated on day 0 with 1,200 embryonated T. canis eggs. Fifteen days before the experimental infection, group 1 (control) was treated via intraperitoneal injection (IP) with sterile distilled water and groups 2 and 3 were treated with dexamethasone (DEX) at 1 and 5 mg/kg/day, respectively. Additionally, group 4 was treated IP with cyclophosphamide (CY) at 50 mg/kg at two times per week for 2 weeks. Sixty days following infection, the mice were euthanised to recover the larvae by means of the tissue digestion technique. The levels of antibodies detected by indirect ELISA were not associated with the larval burden. Administration of CY (50 mg/kg) and DEX (5 mg/kg) resulted in an increase of the larval burden of 162.1% and 50.8%, respectively, in relation to the control group. These two treatments, especially CY (50 mg/kg), promoted immunosuppression and the establishment of a significant larval burden, supporting its further utilisation in studies related to immunosuppression in visceral toxocariasis.

The close contact between dogs and humans poses a high risk of exposure to Toxocara canis eggs, which can lead to Visceral Larva Migrans (VLM) syndrome. The objective of this study was to investigate the existence of these nematode eggs on the hair of the perianal region in stray and owned dogs. Samples of hair from 104 dogs of different ages were collected: 25 (24%) were positive for T. canis eggs, with mean of 614.8 eggs per gram of hair. Puppies were responsible for 99% of the total number of eggs. The ages of the animal and hair length were factors that influenced the intensity of the observed eggs. This study showed that dog hair contaminated by T. canis eggs in different developmental stages represents a potential source of VLM infection for humans., (Copyright © 2010 Elsevier B.V. All rights reserved.)

This study identified pathogenicity genes in 40 Helicobacter pylori clinical isolates. The cagA, vacA, and iceA genes were detected in 65%, 97.5%, and 97.5% of the isolates, respectively. The cagA, iceA1, and vacAs1a/m1 genes were related to erosive gastritis, whereas the vacAs2/m2 and iceA2 genes were associated with enanthematous gastritis.

The increase of incidence of tuberculosis (TB) with resistant strains and HIV co-infection has reinforced the necessity of developing new drugs for its treatment. The reaction of naphthoquinones with aromatic or aliphatic aldehydes in the presence of ammonium acetate led to the synthesis of the three β-lapachone derivatives (naphthoimidazoles) that were tested in this study. Phenazines were prepared by the reaction of the respective naphtoquinone with o-phenylenediamine in acetic acid under reflux. The antimicrobial activity of the derivatives was evaluated in vitro against Mycobacterium tuberculosis H37Rv (ATCC 27294) and the rifampicin-resistant strain (ATCC 35338) containing a His-526-Tir mutation in the rpoB gene. Using the Resazurin Microtiter Assay (REMA) method, bioactive molecules were observed in the susceptible and resistant strains with MICs ranging from 2.2 μM to 17 μM. The naphthoimidazoles with p-toluyl and indolyl group attached to the imidazole ring were more active against the H37Rv strain (MIC 9.12 μM and 4.2 μM, respectively) than the rifampicin-resistant strain (MIC 8.3 μM and 17 μM, respectively). The phenazine with the allyl-pyran group was most active among the two strains and had an MIC of 2.2 mM. These results show the potential of these molecules as prototypes for future drugs used in treating TB.

Aims: This research evaluated the utilization of a urease in-house test, culture and molecular method (ureA PCR) as a diagnostic tool for Helicobacter pylori infection. Furthermore, we assessed the presence of the cagA gene in the specimens and in isolated strains that were positive for ureA by PCR positive., Results: Sensitivity and specificity, respectively, were 100 and 95.8% for the urease in-house test 93.3 and 95.8 for the ureA PCR assay of the specimen and 100 and 100% for the culture. The presence of the cagA gene was observed in eight (53%) ureA-positive samples., Conclusions: In this study, we found that the PCR technique has applicability in the study of cagA, and other genes related to the H. pylori pathogen. This method can be applied to samples directly from biopsy or isolated from the bacteria.

Genotyping tools have been widely used to study the occurrence of outbreaks and to identify the patterns of transmission of Mycobacterium tuberculosis strains. The clonal diversity of 65 clinical isolates of M. tuberculosis was determined by PCR methods. The Double Repeat Element method (DRE-PCR) and spoligotyping identified 45 and 26 distinct patterns respectively. Among these, LAM (38%) was the most frequent lineage, followed by Haarlem (31%) and T (20%). Five orphan patterns were not present in the SITVIT database. Mycobacterial interspersed repetitive units (MIRU) using 12 loci revealed 46 distinct patterns. MIRU loci 10, 23, 26 and 40 had the highest discriminatory power. The high genetic diversity found among M. tuberculosis isolates in this study suggests a high level of recent TB transmission, indicating an endemic mode of TB transmission and a putative importation of new TB genotypes. In addition, the high diversity among the isolates could indicate early detection of the infection in patients and an efficient rate of cure.

Objectives: To evaluate the performance of laboratory diagnosis of tuberculosis, clinical samples underwent culture, species identification and drug susceptibility testing (DST)., Methods: A total of 554 samples from 269 patients were tested for smear microscopy using Kinyoun stain. Culture was performed in Ogawa-Kudoh medium and species identification was performed using the IS6110 amplified region. DST for rifampicin, isoniazid (INH) and streptomycin were carried out using the Resazurin assay., Results: Cultures augmented the number of cases diagnosed by 22.1%, IS6110 amplification identified all Mycobacterium tuberculosis strains thus isolated and DST detected three strains resistant to INH and one multidrug-resistant strain., Conclusion: Simultaneous use of different techniques enhanced culture yield, species identification and detection of drug resistance even in a laboratory with limited facilities.

Tuberculosis remains as a serious infection disease of worldwide distribution, with high morbidity and mortality, mainly in low socio-economic condition countries. The state of emergency of tuberculosis caused by the resistant and multidrug-resistant (MDR) strains, became the main threat to the tuberculosis treatment and control programs. A fast detection method for the resistant strains will allow the implementation of an adequate treatment and contribute for controlling the dissemination of these resistant strains. This study evaluated the performance of the nitrate reductase assay in solid (NRA-LJ) and liquid (NRA-7H9) media, to determine the susceptibility to first line anti-tuberculosis drugs: isoniazid (INH), rifampicin (RMP), ethambutol (EMB) and streptomycin (SMR). Both methods NRA-LJ and NRA-7H9 were evaluated among 18 strains with a known susceptibility profile. The resazurin microtiter assay (REMA) was performed as a reference method. One hundred percent of accordance was observed between NRA-7H9 and REMA for the four tested drugs. When the NRA-LJ method was compared to REMA, the sensitivity and the specificity to INH, RMP, EMB and SMR were 100%, 100 %, 85.7%, 76.9% and 80%, 100%, 75% and 80%, respectively. From the 57 clinical isolates of M. tuberculosis evaluated by NRA-7H9 and REMA, 56 (98.2%) were sensitive to all antibiotics tested (INH, RMP, EMB and SMR) by the NRA-7H9 method, while three of these strains were resistant to INH by REMA. One strain showed resistance to INH and RMP for both methods, and MIC of 1.0 μg/ml to INH for both methods, while MIC of 1.0 and 2.0 μg/ml to RMP for REMA and NRA-7H9, respectively. The three assays showed a high level of agreement for rapid detection of rifampicin and isoniazid resistance. Regarding rapidness, the detection of color change in the NRA method is within instants as compared to the overnight incubation required for the REMA test. NRA might represent an inexpensive and alternative assay for rapid detection of resistance in low-income countries.

In order to investigate the presence of helminths agents of parasitic zoonoses, were examined 237 fecal samples of dogs from central area of the main beach of south coast of Rio Grande do Sul State. The eggs and larvae of genus Ancylostoma (71.3%) and Trichuris (32.5%) and Toxocara eggs (9.3%) were the most prevalent parasites detected.