LARVICIDE ACTIVITY OF LACTOBACILLUS SPP. AND SACCHAROMYCES BOULARDII SUPERNATANTS ON TOXOCARA CANIS.

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.

Saccharomyces boulardii reduces the vertical transmission of Toxocara canis larvae in mice.

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.

Predictive Value of CD8 Expression and FoxP3 Methylation in Nasopharyngeal Carcinoma Patients Treated with Chemoradiotherapy in a Non-endemic Area.

Undifferentiated Nasopharyngeal Carcinoma (UNPC) is associated with Epstein-Barr Virus (EBV) and characterized by an abundant immune infiltrate potentially influencing the prognosis. Thus, we retrospectively assessed the significance of immunosuppression in the UNPC microenvironment as prognostic biomarker of treatment failure in a non-endemic area, and monitored the variation of systemic EBV-specific immunity before and after chemoradiotherapy (CRT). DNA and RNA were extracted from diagnostic biopsies obtained by tumor and adjacent mucosa from 63 consecutive EBV+ UNPC patients who underwent radical CRT. Among these patients 11 relapsed within 2 years. The expression of the EBV-derived UNPC-specific BARF1 gene and several immune-related genes was monitored through quantitative RT-PCR and methylation-specific PCR analyses. Peripheral T cell responses against EBV and BARF1 were measured in 14 patients (7 relapses) through IFN-γ ELISPOT assay. We found significantly higher expression levels of BARF1, CD8, IFN-γ, IDO, PD-L1, and PD-1 in UNPC samples compared to healthy tissues. CD8 expression was significantly reduced in both tumor and healthy tissues in UNPC patients who relapsed within two years. We observed a hypomethylated FOXP3 intron 1 exclusively in relapsed UNPC patients. Finally, we noticed a significant decrease in EBV- and BARF1-specific T-cells after CRT only in relapsing patients. Our data suggest that a high level of immunosuppression (low CD8, hypomethylated FoxP3) in UNPC microenvironment may predict treatment failure and may allow an early identification of patients who could benefit from the addition of immune modulating strategies to improve first line CRT.

Frequency of enteroparasitic infections and serum positivity for Toxocara spp. in children from a public day care center in Southern Brazil / Frequência de enteroparasitos e de soropositividade para Toxocara spp. em crianças de uma creche pública do Sul do Brasil

Abstract 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.

Resumo Introdução As creches desempenham um importante papel social na educação e desenvolvimento inicial das crianças. O objetivo deste estudo foi investigar a frequência de infecções parasitárias intestinais, além de infecção por Toxocara spp. em crianças de uma creche na cidade do Rio Grande, Rio Grande do Sul, Brasil. Métodos: Para o estudo, foram coletadas amostras fecais de 50 crianças de três a seis anos, analisadas pelas técnicas de Ritchie, Faust, Rugai e Kinyoun, sendo aplicado um questionário epidemiológico fechado e estruturado aos pais/responsáveis ​​das crianças. Também foi realizada uma pesquisa sorológica de anticorpos para Toxocara spp. em 41 crianças, pelo ensaio imunoenzimático, associado ao antígeno de excreção e secreção (ES), com soros pré-adsorvidos com antígeno somático de Ascaris lumbricoides. Resultados: A frequência de enteroparasitos foi de 18%, enquanto que 43.9% das crianças foram soropositivas para Toxocara spp.. Baixa renda familiar e baixo nível de alfabetização dos pais/responsáveis ​​foram fatores comuns entre as famílias das crianças parasitadas. A maioria dos pais ou responsáveis das crianças, com diagnóstico positivo para enteroparasitos ou para Toxocara spp., relatou utilizar apenas água para a higienização de verduras e frutas. Conclusões: As altas taxas de soropositividade para Toxocara spp. indicam que as crianças foram expostas a esse parasito, sendo importante minimizar o risco de infecção. Além disso, a positividade de 18% para enteroparasitos e os dados obtidos com as entrevistas realizadas com os pais ou responsáveis demonstram a necessidade da realização de trabalhos com a população estudada que visem a divulgação sobre as parasitoses intestinais, especialmente sobre medidas profiláticas especificas.

Frequency of enteroparasitic infections and serum positivity for Toxocara spp. in children from a public day care center in Southern Brazil.

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.

Risk and other factors associated with toxoplasmosis and toxocariasis in pregnant women from southern Brazil.

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.

Frequency of enteroparasitic infections and serum positivity for Toxocara spp. in children from a public day care center in Southern Brazil

Abstract Introduction Day care centers play an important social role in childrens 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 Kinyouns 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.

Resumo Introdução As creches desempenham um importante papel social na educação e desenvolvimento inicial das crianças. O objetivo deste estudo foi investigar a frequência de infecções parasitárias intestinais, além de infecção por Toxocara spp. em crianças de uma creche na cidade do Rio Grande, Rio Grande do Sul, Brasil. Métodos: Para o estudo, foram coletadas amostras fecais de 50 crianças de três a seis anos, analisadas pelas técnicas de Ritchie, Faust, Rugai e Kinyoun, sendo aplicado um questionário epidemiológico fechado e estruturado aos pais/responsáveis das crianças. Também foi realizada uma pesquisa sorológica de anticorpos para Toxocara spp. em 41 crianças, pelo ensaio imunoenzimático, associado ao antígeno de excreção e secreção (ES), com soros pré-adsorvidos com antígeno somático de Ascaris lumbricoides. Resultados: A frequência de enteroparasitos foi de 18%, enquanto que 43.9% das crianças foram soropositivas para Toxocara spp.. Baixa renda familiar e baixo nível de alfabetização dos pais/responsáveis foram fatores comuns entre as famílias das crianças parasitadas. A maioria dos pais ou responsáveis das crianças, com diagnóstico positivo para enteroparasitos ou para Toxocara spp., relatou utilizar apenas água para a higienização de verduras e frutas. Conclusões: As altas taxas de soropositividade para Toxocara spp. indicam que as crianças foram expostas a esse parasito, sendo importante minimizar o risco de infecção. Além disso, a positividade de 18% para enteroparasitos e os dados obtidos com as entrevistas realizadas com os pais ou responsáveis demonstram a necessidade da realização de trabalhos com a população estudada que visem a divulgação sobre as parasitoses intestinais, especialmente sobre medidas profiláticas especificas.

Modulation of IL-12 and IFNγ by probiotic supplementation promotes protection against Toxocara canis infection in mice.

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.

Toxocara canis: anthelmintic activity of quinone derivatives in murine toxocarosis.

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.

Transcriptome modification of white blood cells after dietary administration of curcumin and non-steroidal anti-inflammatory drug in osteoarthritic affected dogs.

The dietary effect of non-steroidal anti-inflammatory drug (NSAID) or curcumin on the gene expression of peripheral white blood cells in osteoarthritis (OA) affected dogs was investigated using a 44K oligo microarray. Two groups of OA dogs and one group of healthy dogs (6 dogs each) were clinically evaluated and blood was sampled before (T0) and after 20days (T20) of dietary administration of NSAID (NSAID group) or curcumin (CURCUMIN group). Differentially expressed genes (P<0.05) in comparison to the control group were identified with MeV software and were functional annotated and monitored for signaling pathways and candidate biomarkers using the Ingenuity Pathways Analysis (IPA). After 20days of treatment, the differentially expressed transcripts significantly (P<0.05) decreased from 475 to 173 in NSAID group and from 498 to 141 in CURCUMIN group. Genes involved in "inflammatory response" and in "connective tissue development and function" dramatically decreased at T20. Other genes, included in "cellular movement", "cellular compromise" and "immune cell trafficking", were differentially expressed at T0 but not at T20 in both groups. Specific molecular targets of CURCUMIN, not observed for NSAID, were the IkB up regulation in the "TNRF1 signaling pathway" and IL18 down regulation in the "role of cytokines in mediating communication between immune cells". The activity of CURCUMIN was also evidenced from the inhibition of macrophages proliferation (HBEGF), related to a strong down regulation of TNFα and to activation of fibrinolysis (SERPINE1). The results would suggest that curcumin offers a complementary antinflammatory support for OA treatment in dogs.

The laboratory as a tool to qualify tuberculosis diagnosis.

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.