RONALDO CESAR BORGES GRYSCHEK
Projetos de Pesquisa
Unidades Organizacionais
Departamento de Moléstias Infecciosas e Parasitárias, Faculdade de Medicina - Docente
LIM/06 - Laboratório de Imunopatologia da Esquistossomose e outras Parasitoses, Hospital das Clínicas, Faculdade de Medicina
LIM/06 - Laboratório de Imunopatologia da Esquistossomose e outras Parasitoses, Hospital das Clínicas, Faculdade de Medicina
11 resultados
Resultados de Busca
Agora exibindo 1 - 10 de 11
- Seroprevalence and associated risk factors of strongyloidiasis in indigenous communities and healthcare professionals from Brazil(2023) SANTAREM, Vamilton Alvares; DOLINE, Fernando Rodrigo; SANTOS, Joao Henrique Farinhas dos; FERREIRA, Isabella Braghin; GOMES, Bruna Barroso; MEISEL, Dirce Mary Correa; BIONDO, Leandro Meneguelli; LESCANO, Susana Angelica Zevallos; GRYSCHEK, Ronaldo Cesar Borges; GIUFFRIDA, Rogerio; SANTOS, Andrea Pires dos; KMETIUK, Louise Bach; PAULA, Fabiana Martins de; BIONDO, Alexander WelkerAuthor summaryHuman strongyloidiasis, caused mainly by nematoda Strongyloides stercoralis, with estimate of infecting around 613.9 million people worldwide, and may play a pivotal role in causing morbidity in aboriginals and indigenous populations worldwide. Nevertheless, prevalence and risk factors for Strongyloides stercoralis in Brazilian indigenous populations remains to be fully established. The present study was the first comparative report of S. stercoralis seroprevalence in indigenous population and healthcare professionals. A high seroprevalence to Strongyloides stercoralis in indigenous communities of Brazil along with even higher exposure of healthcare professionals was reported and should be considered as vulnerability and health worker exposure. Male and adult indigenous persons were more likely seropositive as associated risk factors, while having septic tank as a sanitary facility represented a protective factor for S. stercoralis exposure. Thus, the present study has alerted for the public health concerns of strongyloidiasis in such vulnerable populations, particularly when lacking sanitation conditions. Finally, a further One Health approach may contribute for better understanding the strongyloidiasis exposure including the environmental, animal, and human components to battle this important parasitic neglected disease in indigenous communities. Strongyloides stercoralis, a pathogenic roundworm, is considered endemic in several tropical and subtropical areas worldwide. Indigenous populations have the highest soil-transmitted helminthiases-related mortality rates, but the prevalence and risk factors associated with S. stercoralis in Brazilian indigenous populations have not been established. Thus, the present study aimed to assess the seroprevalence and associated risk factors for S. stercoralis in indigenous communities and the healthcare professionals serving them in Brazil. Indigenous populations living in nine communities and healthcare professionals were tested for anti- S. stercoralis antibodies by ELISA. A questionnaire was used to assess socio-epidemiological information. Associated risk factors for seropositivity were tested by chi-square or Fisher's exact tests, using bivariate analyses and multivariate logistic regression. Overall, 174/463 (37.6%; CI 95%: 33.3-42.1) indigenous persons and 77/147 (52.4%; 95% CI: 44.3-60.3) healthcare professionals were seropositive for anti- S. stercoralis antibodies. Seropositivity among the two groups was statistically significant (p = 0.0016; OR = 0.547; 95% CI: 0.376-0.796) and revealed that healthcare professionals were 1.83 times more likely to be seropositive. The multivariate analysis showed that being male or being adult were also risk factors, while having a septic tank as a sanitary facility represented a protective factor for S. stercoralis exposure in indigenous persons. None of the variables evaluated were associated with S. stercoralis exposure in the professional group. The study herein has reported a high seroprevalence to Strongyloides stercoralis in indigenous communities of Brazil and healthcare professionals, warning for potential public health concerns of strongyloidiasis in such populations.
- Immunoreactivity of proteins within 30-40 kDa range during the acute and the recovery phases in rats experimentally infected with Strongyloides venezuelensis(2020) FONSECA, Priscilla Duarte Marques; CORRAL, Marcelo Andreeta; MEISEL, Dirce Mary C. Lima; LEVI, Debora; NASCIMENTO, Rafael Correa; CASTRO-BORGES, William; GRYSCHEK, Ronaldo; COSTA-CRUZ, Julia Maria; PAULA, Fabiana Martins deIn experimental infection with Strongyloides venezuelensis, the acute and recovery phases can be distinguished, unlike human infections caused by Strongyloides stercoralis. The objective of this study was to evaluate the production of anti-Strongyloides IgG antibodies and the recognition of immunogenic protein bands during the acute and the recovery phases in rats experimentally infected with S. venezuelensis. Rats were infected subcutaneously with 400 or 4,000 S. venezuelensis infective larvae. The acute phase was characterized by elimination of a large number of eggs in the faeces on days 6-14 post infection; the recovery phase was characterized by the resolution of the infection between days 30 and 35 post infection. Differences in IgG levels were observed in the acute and the recovery phases. Different antigenic fractions were recognized in both phases of infection. It is concluded that proteins within the 30-40 kDa range are immunoreactive markers for both the acute and the recovery phases in rats experimentally infected with S. venezuelensis, particularly using membrane antigen.
- Current status of research regarding Blastocystis sp., an enigmatic protist, in Brazil(2021) MELO, Gessica Baptista de; BOSQUI, Larissa Rodrigues; COSTA, Idessania Nazareth da; PAULA, Fabiana Martins de; GRYSCHEK, Ronaldo Cesar BorgesThe present study aimed to evaluate the occurrence of Blastocystis sp. in Brazilian studies over a period of years (2000-2020), as well as point out relevant aspects of this enigmatic organism. We performed a literature search using six sources of international databases. The data were divided into diagnostic by parasitological and molecular techniques, and relevant aspects. After applying the inclusion and exclusion criteria, 52 studies were included in the final analysis. The occurrence of Blastocystis sp. in Brazil ranged from 0.5% to 86.6%, as determined using parasitological techniques. The highest occurrence was in the North (27.3%) and the lowest, in the Midwest region (13.4%). In Brazil, most studies have employed molecular techniques and are concentrated in the Southeast region. The Blastocystis sp. subtype ST3 had the highest average positivity, followed by ST1 and ST2. These findings represent a panorama that reflects the reality of Brazil; thus, we believe that the effectiveness of parasitological diagnosis should be considered with regard to making an appropriate choice of technique for detecting Blastocystis sp. Additionally, we emphasize the importance of further studies in the context of molecular epidemiology with regard to this genus. Blastocystis sp. is not well understood yet, and very little information regarding this genus is available; hence, further research regarding this genus is urgently needed.
- Toxocara canis 30-35 kDa excretory-secretory antigen is an important marker in mice challenged by inocula containing different parasite load levels(2022) FONSECA, Gabriela Rodrigues e; CORRAL, Marcelo Andreetta; PAULA, Fabiana Martins de; MEISEL, Dirce Mary Correia Lima; GRYSCHEK, Ronaldo Cesar Borges; LESCANO, Susana Angelica ZevallosThe Western-blotting technique was applied to identify antigenic fractions of excretory-secretory Toxocara canis antigen recognized by IgG antibodies throughout an experimental infection in mice challenged by different inocula. Mice were inoculated with 5, 50 and 500 embryonated eggs and serum samples were collected 15, 30, 60, 90 and 120 days post-infection. Serum samples were analyzed using an excretory-secretory Toxocara antigen. Antibodies recognized antigenic fractions from 30 to 90 kDa. The protein fraction of 30-35 kDa was the most frequently recognized regardless of the size of inoculum and the stage of infection represented by the different collection times, but the antigenic recognition was more evident in groups infected with 50 and 500 eggs. This study presents an antigenic panel of the excretory secretory antigen of T. canis and suggests that the 30-35 kDa antigenic fraction is a promising marker of the infection and should be further explored in future studies on experimental toxocariasis.
- Screening of Strongyloides infection using an ELISA test in transplant candidates(2019) TOLEDO, Beatriz; CORRAL, Marcelo A.; MEISEL, Dirce Mary C. L.; GOTTARDI, Maiara; ABDALA, Edson; COSTA, Silvia F.; PIERROTTI, Ligia Camera; LESCANO, Susana A. Z.; GONCALVES, Elenice M. N.; CASTILHO, Vera L. P.; CHIEFFI, Pedro P.; GRYSCHEK, Ronaldo C. B.; PAULA, Fabiana M.OBJECTIVES: Hyperinfection or disseminated strongyloidiasis has been frequently reported after transplants and is related to high mortality. This study aimed to screen for strongyloidiasis using serological diagnoses in transplant candidates. METHODS: An ELISA test was performed with filariform larvae of Strongyloides venezuelensis as a source of antigen. RESULTS: In the serum from transplant candidates, anti-Strongyloides IgG antibodies were detected in 35/150 (23.3%) samples by soluble fractions in phosphate buffered saline (PBS), 31/150 (20.7%) samples by soluble fractions in Tris-HCl, 27/150 (18.0%) samples by membrane fractions in PBS and 22/150 (14.7%) samples by membrane fractions in Tris-HCl. CONCLUSIONS: The present results suggest the ELISA test, ideally using soluble fractions of filariform larvae S. venezuelensis in PBS, as an additional strategy for the diagnosis of strongyloidiasis in transplant candidates.
- IMMUNODIAGNOSIS OF HUMAN STRONGYLOIDIASIS: USE OF SIX DIFFERENT ANTIGENIC FRACTIONS FROM Strongyloides venezuelensis PARASITIC FEMALES(2015) CORRAL, Marcelo Andreetta; PAULA, Fabiana Martins de; GOTTARDI, Maiara; MEISEL, Dirce Mary Correia Lima; CASTILHO, Vera Lucia Pagliusi; GONCALVES, Elenice Messias do Nascimento; CHIEFFI, Pedro Paulo; GRYSCHEK, Ronaldo Cesar BorgesThe aim of this study was to evaluate six different antigenic fractions from Strongyloides venezuelensis parasitic females for the immunodiagnosis of human strongyloidiasis. Soluble and membrane fractions from S. venezuelensis parasitic females were prepared in phosphate-buffered saline (SSF and SMF, respectively), Tris-HCl (TSF and TMF, respectively), and an alkaline buffer (ASF and AMF, respectively). Serum samples obtained from patients with strongyloidiasis or, other parasitic diseases, and healthy individuals were analyzed by enzyme-linked immunosorbent assay (ELISA). Soluble fractions SSF, TSF, and ASF showed 85.0%, 75.0%, and 80.0% sensitivity and 93.1%, 93.1%, and 87.5% specificity, respectively. Membrane fractions SMF, TMF, and AMF showed 80.0%, 75.0%, and 85.0% sensitivity, and 95.8%, 90.3%, and 91.7% specificity, respectively. In conclusion, the present results suggest that the fractions obtained from parasitic females, especially the SSF and SMF, could be used as alternative antigen sources in the serodiagnosis of human strongyloidiasis.
- Immune complexes as a tool for strongyloidiasis immunodiagnosis in kidney and liver transplant candidate(2022) CORRAL, Marcelo A.; GONCALVES, Ana Lucia R.; COSTA, Idessania N.; ABDALA, Edson; PIERROTTI, Ligia C.; CHIEFFI, Pedro Paulo; COSTA-CRUZ, Julia Maria; GRYSCHEK, Ronaldo Cesar B.; PAULA, Fabiana MartinsStrongyloidiasis is a chronic and asymptomatic infection in immunocompetent patients. Immunocompromised patients, such as organ transplant candidates, can develop severe forms of this disease, and the best way to prevent progression to these forms is early diagnosis. Serological techniques using specific IgG and immune complexes (IC) detection can help in the diagnosis of these patients. This study aimed to detect specific anti-Strongyloides IC and IgG antibodies in kidney transplant (KT) and liver transplant (LT) candidates. A total of 100 blood samples was collected from transplant candidates (50 blood samples each from KT and LT candidates). Serum was obtained and analysed using enzyme-linked immunosorbent assay for IC and IgG detections. The IC levels showed frequencies of 18% and 2% in the KT and LT groups, respectively, whereas anti-Strongyloides IgG was detected in 34% and 12% of KT and LT candidates, respectively. The correlation between IC and IgG detection is poor in KT candidates, while in LT candidates, there is a significant positive correlation. The detection of IC can be an additional tool for the diagnosis of strongyloidiasis, especially when associated with the detection of specific IgG anti-Strongyloides antibodies.
- IgG reactivity with 40-35 kDa soluble and membrane antigen of Strongyloides venezuelensis in immunocompromised patients(2019) CORRAL, Marcelo Andreetta; PAULA, Fabiana Martins de; MEISEL, Dirce Mary C. L.; ABDALA, Edson; COSTA, Silvia Figueiredo; PIERROTTI, Ligia Camera; YAMASHIRO, Juliana; GONCALVES, Elenice M. do Nascimento; CASTILHO, Vera Lucia P.; CHIEFFI, Pedro Paulo; GRYSCHEK, Ronald Cesar B.Immunocompromised patients constitute a risk group for the development of severe clinical forms of human strongyloidiasis. The diagnosis of this infection is primarily performed by parasitological techniques, but with low sensitivity. Serological techniques appear as an alternative, especially with heterologous antigens use. The aim of this study was to perform the Western blot technique by using S. venezuelensis infective third stage larva (iL3) soluble (TS) and membrane (TM) saline antigens to reveal immunoreactive bands in immunocompromised patients with strongyloidiasis. Serum samples from 117 parasitologically well-characterized patients were divided into four groups: S. stercoralis positive and immunocompetent (S + IC); S. stercoralis positive and immunocompromised (S + IP); negative and immunocompetent (S-IC); negative and immunocompromised (S-IP). A 40-35 kDa band was recognized by 100% of patients in the S + IC group in both antigenic fractions, and by 62.5% and 50% in the S + IP group using the TS and TM fractions, respectively. A 29 kDa band was recognized by 86.3% and 72.7% (for TS and TM, respectively) of patients in the S + IC group, and only by 12.5% of patients in the S + IP group on the TM antigen. Regardless of the patients' immunological condition, the 40-35 kDa band from S. venezuelensis was detected more frequently and can be used as an important marker to the immunodiagnosis of human strongyloidiasis.
- Evaluation of real-time PCR assay to detect Schistosoma mansoni infections in a low endemic setting(2014) ESPIRITO-SANTO, Maria Cristina Carvalho; ALVARADO-MORA, Monica Viviana; DIAS-NETO, Emmanuel; BOTELHO-LIMA, Livia Souza; MOREIRA, Joao Paulo; AMORIM, Maria; PINTO, Pedro Luiz Silva; HEATH, Ashley R.; CASTILHO, Vera Lucia Pagliusi; GONCALVES, Elenice Messias do Nascimento; LUNA, Expedito Jose de Albuquerque; CARRILHO, Flair Jose; PINHO, Joao Renato Rebello; GRYSCHEK, Ronaldo Cesar BorgesBackground: Schistosomiasis constitutes a major public health problem, and 200 million people are estimated to be infected with schistosomiasis worldwide. In Brazil, schistosomiasis has been reported in 19 states, showing areas of high and medium endemicity and a wide range of areas of low endemicity (ALE). Barra Mansa in Rio de Janeiro state has an estimated prevalence of 1%. ALE represent a new challenge for the helminth control because about 75% of infected individuals are asymptomatic and infections occur with a low parasite load (<100 eggs per gram of feces), causing a decrease in sensitivity of stool parasitological techniques, which are a reference for the laboratory diagnosis of this helminth. The objective of this study was to evaluate the performance of a TaqMan quantitative polymerase chain reaction (qPCR) technique in serum and feces DNA samples using the techniques of Kato-Katz (KK), Hoffman, Pons and Janer (HH) as references, during an epidemiological survey using fecal samples and sera from randomized residents from an ALE. Methods: A cross-sectional study conducted from April to December 2011 using a probabilistic sampling that collected 572 fecal and serum samples. The laboratory diagnostic techniques used were: KK, HH and qPCR ( feces and serum). Results: We obtained the following results using the different diagnostic techniques: KK and HH, 0.9% (n = 5); qPCR-feces, 9.6% (n = 55); and qPCR-serum, 1.4% (n = 8). The qPCR-feces presented the highest positivity, whereas the techniques of HH and KK were the least sensitive to detect infections (0.8%). Compared to HH and KK, qPCR-feces showed a statistically significant difference in positivity (p < 0.05), although with poor agreement. Conclusion: The positivity rate presented by the qPCR approach was far higher than that obtained by parasitological techniques. The lack of adequate surveillance in ALE of schistosomiasis indicates a high possibility of these areas being actually of medium and high endemicity. This study presents a control perspective, pointing to the possibility of using combined laboratory tools in the diagnosis of schistosomiasis in ALE.
- Detection of Schistosoma mansoni infection by TaqMan (R) Real-Time PCR in a hamster model(2014) ESPIRITO-SANTO, Maria Cristina Carvalho; ALVARADO-MORA, Monica Viviana; PINTO, Pedro Luiz Silva; BRITO, Thales de; BOTELHO-LIMA, Livia; HEATH, Ashley Richard; AMORIM, Maria Galli; DIAS-NETO, Emmanuel; CHIEFFI, Pedro Paulo; PINHO, Joao Renato Rebello; CARRILHO, Flair Jose; LUNA, Expedito Jose Albuquerque; GRYSCHEK, Ronaldo Cesar BorgesAn experimental study in hamsters was performed to evaluate the capability for detecting Schistosoma mansoni DNA in serum and fecal samples during the pre and post-egg-laying periods of infection using TaqMan (R) Real-Time PCR system (qPCR), was compared with the circumoval precipitin test (COPT) and the Kato-Katz technique, especially among individuals with low parasitic burden. Twenty-four hamsters were infected with cercariae. Three hamsters were sacrificed per week under anesthesia, from 7 days post infection (DPI) up to 56 DPI. A serum sample and a pool of feces were collected from each hamster. The presence of S. mansoni eggs in fecal samples was evaluated by Kato-Katz method and in the hamsters gut-by histopathology. Detection of S. mansoni DNA was performed using qPCR and S. mansoni antibody using COPT. The first detection of eggs in feces by Kato-Katz method and S. mansoni DNA in feces by qPCR occurred 49 DPI. Nevertheless, S. mansoni DNA was detected in serum samples from 14 up to 56 DPI. COPT was positive at 35 DPI. The results not only confirm the reliability of S. mansoni DNA detection by qPCR, but also demonstrate that serum is a trustworthy source of DNA in the pre patent infection period.