Molecular characterization and epidemiology of Toxoplasma gondii isolates from free-range chickens in the southwest region of Goiás: new genotypes.

The purpose of this study was to isolate Toxoplasma gondii from tissues of free-range chickens in the southwestern region of Goiás, to detect and molecularly characterize the genetic material of the parasite, and to determine the seroprevalence of the protozoan parasite in these animals. A seroprevalence of T. gondii antibodies of 76% (19/25) was found among the chickens, while genetic material from their tissues was detected in 56% (14/25). A total of 14 isolates was obtained in the bioassay, ten of which were considered acute, eight were considered isolates of high virulence lethal to mice, and four of low virulence, considered non-lethal but with the ability to chronify the infection. Seven of the ten isolates showed significant morphometric differences from the RH strain, in terms of nucleus-complex-apical distance, length and width. Genotyping of the acute isolates was performed by RFLP-PCR, using 11 genetic markers: SAG1, SAG2 (3'SAG2 and 5'SAG2), alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and APICO. The results were compared and classified according to the genotypes listed on the ToxoDB Platform, where different profiles were observed indicating the presence of two known genotypes (#7 and #63) and five new genotypes (NEW 3, NEW4, NEW5, NEW6, NEW 7). The results showed high seroprevalence, isolation rate, molecular detection and genotypic variations of T. gondii in free-range chickens in the southwestern region of Goiás.

IgG Avidity in Samples Collected on Filter Paper: Importance of The Early Diagnosis of Congenital Toxoplasmosis.

OBJECTIVE: The purpose of the present study is to standardize and evaluate the use of the immunoglobulin G (IgG) antibody avidity test on blood samples from newborns collected on filter paper to perform the heel test aiming at its implementation in ongoing programs. METHODS: Blood samples from newborns were collected on filter paper simultaneously with the heel prick test. All samples were subjected to immunoglobulin M IgM and IgG enzyme-linked immunosorbent assays (ELISA). Peripheral blood was collected again in the traditional way and on filter paper from newborns with high IgG levels (33). Three types of techniques were performed, the standard for measuring IgG in serum, adapted for filter paper and the technique of IgG avidity in serum and on filter paper. The results of the avidity test were classified according to the Rahbari protocol. RESULTS: Among the 177 samples, 17 were collected in duplicate from the same child, 1 of peripheral blood and 1 on filter paper. In this analysis, 1 (5.88%) of the 17 samples collected in duplicate also exhibited low IgG avidity, suggesting congenital infection. In addition, the results obtained from serum and filter paper were in agreement, that is, 16 (94.12%) samples presented high avidity, with 100% agreement between the results obtained from serum and from filter paper. CONCLUSION: The results of the present study indicate that the avidity test may be another valuable method for the diagnosis of congenital toxoplasmosis in newborns.

OBJETIVO: O objetivo do presente estudo é padronizar e avaliar a utilização do teste de avidez de anticorpos imunoglobulina G (IgG) em amostras de sangue de recém-nascidos (RNs) coletadas em papel filtro para a realização do teste do pezinho visando a implementação nos programas já vigentes. MéTODOS: Foram coletadas amostras de sangue de recém-nascidos em papel filtro simultaneamente ao teste do pezinho. Em todas as amostras, foram realizados os testes imunoenzimáticos (ELISA) imunoglobulina M (IgM) e IgG. Dos RNs que apresentaram altos índices de IgG (33), foi novamente coletado sangue periférico da forma tradicional e em papel filtro. Foram realizadas técnicas padrão para a dosagem de IgG em soro, adaptadas para papel filtro, e a técnica de avidez de IgG em soro e em papel filtro. Os valores obtidos para o teste de avidez foram classificados de acordo com o protocolo de Rahbari. RESULTADOS: Dentre as 177 recoletas, em 17 amostras foi realizada a coleta simultânea de sangue periférico e papel filtro da mesma criança. Nesta análise, 1 (5,88%) das 17 amostras coletadas em duplicata obteve também baixa avidez de IgG, sugerindo infecção congênita da criança, e houve concordância entre os resultados obtidos em soro e em papel filtro: 16 (94,12%) das amostras apresentaram alta avidez, com concordância de 100% entre os resultados obtidos em soro e em papel filtro. CONCLUSãO: Os dados do presente trabalho evidenciam que o teste de avidez poderá ser mais um método valioso a ser utilizado no diagnóstico da toxoplasmose congênita em RNs.

Copro-PCR in the detection and confirmation of Toxoplasma gondii oocysts in feces of stray and domiciled cats.

Molecular methods such as Copro-PCR stand out in the diagnosis of T. gondii, because they are highly sensitive and specific, and can distinguish T. gondii from other morphologically similar coccids. The purpose was the detection of Toxoplasma gondii copro-prevalence by polymerase chain reaction in 149 fecal samples from stray and domiciled cats, using three distinct markers (B5-B6, 18S and 529bp RE). Oocysts of T. gondii/H. hammondi were detected in 15.4% by parasitology fecal tests (PFT), and 4% of these oocysts were positively identified as T. gondii by Copro-PCR. The presence of T. gondii genetic material was detected in 16.1%, but 12% of the samples that tested positive by Copro-PCR were negative in PFT. Samples with discordant results were subjected to a new Copro-PCR with 18S marker and a 529, and of the 17 samples, 9 contained T. gondii genetic material. A comparison of the PFT and the molecular methods showed the latter was more sensitive, since it detected 22.1% while the PFT detected 15.4%. Demonstrating the high sensitivity and specificity of the Copro-PCR, particularly with the association of primers (k=0.809), but also confirms the importance of using molecular techniques in laboratories, since Copro-PCR was able to detect samples considered negative by PFT.

Molecular characterization of Toxoplasma gondii isolates from free-range chickens reveals new genotypes in Goiânia, Goiás, Brazil.

The aim of this study was to evaluate the genotypic characteristics of Toxoplasma gondii isolated from free-range chickens in the metropolitan area of Goiânia, Goiás, in Brazil's central-west region. The seroprevalence rate was found to be 96%, according to an indirect hemagglutination assay. Brain and heart samples were processed by peptic digestion for a mice bioassay. The tissues were homogenized and the resulting samples were subjected to polymerase chain reaction (PCR), which revealed that 64% of them contained the parasite's DNA. The mice bioassay revealed 15 isolates, 8 of them tachyzoites isolates from the peritoneal lavage and 7 from brain cysts. T. gondii genotypes were determined through PCR-RFLP, using the following markers: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, alt. SAG2, Apico and CS3. Three genotypes were identified, inclued ToxoDB #65, and the other two are not yet described in the literature. Hence, we conclude that the isolates obtained from the metropolitan area of Goiânia showed relatively low genetic diversity.

Plasmodium falciparum malarial parasites from Brazil lack artemisinin resistance-associated mutations in the kelch13 gene.

INTRODUCTION: Mutations in the propeller domain of the Plasmodium falciparum kelch13 (k13) gene are associated with artemisinin resistance. METHODS: We developed a PCR protocol to sequence the pfk13 gene and determined its sequence in a batch of 50 samples collected from 2003 to 2016 in Brazil. RESULTS: We identified 1 K189T substitution located outside the propeller domain of the PfK13 protein in 36% of samples. CONCLUSIONS: Although the sample size is relatively small, these results suggest that P. falciparum artemisinin-resistant mutants do not exist in Brazil, thereby supporting the continuation of current treatment programs based on artemisinin-based combination therapy.

Basic heel prick test: inclusion of screening, diagnosis and criteria for early confirmation of congenital infection by Toxoplasma gondii.

Toxoplasma gondii can cross the placental barrier, causing fetal infection with potentially severe sequelae. The aim of this study was to evaluate whether the serological screening for toxoplasmosis should be included in the basic neonatal heel prick test in order to establish criteria for the confirmation and/or exclusion of the diagnosis of congenital infection in newborns treated at three public health units in the metropolitan region of Goiania, Goias State, Brazil. Blood samples were collected on filter paper from newborns and later, peripheral blood samples from the mothers and their respective children were obtained to confirm or exclude the diagnosis of suspected congenital infection, by means of an enzyme-linked immunosorbent assay (IgM and IgG) and a polymerase chain reaction assay. From a total of 1,159 blood samples collected on filter paper, 43.92% were reactive to IgG and 0.17% to anti-T. gondii IgM and IgG. One hundred and twenty-seven paired samples (mother and child) were collected following consensual protocols for peripheral blood collection. Results obtained from the filter paper and peripheral blood of the newborns were 90.55% concordant. A comparison of the mother and child blood test results showed agreement regarding the detection of IgG in 90.48% of the samples. The parasite DNA was detected in the peripheral blood of one child. In view of the results obtained in this study, the inclusion of the serological screening for toxoplasmosis in the newborn heel prick test proved to be effective for the early detection of congenital T. gondii infection.

Analysis of the accuracy of different laboratory methods for the diagnosis of intestinal parasites from stray and domiciled cats (Felis catus domesticus) in Goiânia, Goiás, Brazil.

Cats are carriers of zoonotic agents to humans, including intestinal parasites. The purpose of this study was to analyze the accuracy of different laboratory methods for the diagnosis of intestinal parasites. Fecal samples were processed by the Willis, Sheather, Faust and Hoffman-Janer-Pons-Lutz (HJPL) methods. Accuracy analysis was performed determining the sensitivity, specificity, positive and negative predictive value and Kappa. A total of 149 fecal samples were collected, 65 from stray cats and 84 from domiciled cats. The prevalence of intestinal parasites in stray cats was 60% while in domiciled cats it was 17%. In the analysis of accuracy, the techniques that showed the greatest accuracy for Ancylostomids were Willis and Faust, for Cystoisospora spp. Sheather with Faust or HPJL, and Toxoplasma gondii/Hammondia hammondi the association between Willis and Faust. Therefore, for a reliable evaluation of the prevalence of intestinal parasites, at least two different techniques should be used in parasitological exams of feces.

Epidemiological factors associated with Toxoplasma gondii infection in postpartum women treated in the public healthcare system of Goiânia, State of Goiás, Brazil.

INTRODUCTION: Knowledge of the epidemiological profile and risk factors for Toxoplasma gondii infection among postpartum women is a relevant issue, because this protozoan can be vertically transmitted to the developing fetus, which can cause severe and debilitating disease. The aim of this study was to assess the risk factors associated with T. gondii infection in postpartum women in Goiânia, GO, Brazil. METHODS: This cross-sectional study comprised 229 postpartum women, among whom 204 were chronically infected (IgG+/IgM-), and 25 were seronegative (IgG-/IgM-; control group). All the patients were asked to complete a form to provide sociodemographic, clinical, dietary, and cultural information. The data were analyzed to compare seropositivity and risk factors based on the odds ratio (OR) thereof. RESULTS: The sociodemographic characteristics associated with the risk for toxoplasmosis were: education ≤ 8 years [OR: 2.521, confidence interval (CI): 1.01-6.301, p=0.049], and age ≥ 30 years (OR: 4.090; CI: 1.180-14.112, p=0.023). Clinical and behavioral characteristics related to eating raw and undercooked meat, were not found to be risk factors associated with a positive test for toxoplasmosis. CONCLUSIONS: Our findings concur with the results of other studies conducted in Brazil and abroad, where variables such as low levels of schooling, and advanced age (≥ 30 years) are major risk factors for pregnant women to become infected with T. gondii.

The first Acanthamoeba keratitis case in the Midwest region of Brazil: diagnosis, genotyping of the parasite and disease outcome.

We report an Acanthamoeba keratitis case associated with the use of contact lens in a 28-year-old female from Brasília, Brazil. Samples from corneal scraping and contact lens case were used for culture establishment, PCR amplification, and partial sequencing (fragments of ~400kb) of small subunit rDNA; both culture and PCR were positive. The sequence analyses of the cornea and of isolates from the contact lens case showed similarity with the T4 genotype. To the best of our knowledge, this is the first report of T4 Acanthamoeba keratitis case from the Midwest region of Brazil.

Nursing infant with acquired toxoplasmosis in the first months of life - a case report.

Toxoplasmosis is caused by Toxoplasma gondii and the probability of this infection occurring in the first months of life is usually low because its transmission is related to eating habits. A 6-month-old nursing infant was diagnosed with acute toxoplasmosis, which was identified through anti- T. gondii IgA, IgM and low-avidity IgG serologic assays, polymerase chain reaction (PCR) and mouse bioassay test although its mother was seronegative. This serological divergence between mother and child led us to interview the mother regarding epidemiological factors. During this interview, she reported that she had given her 2-month-old baby a piece of undercooked beef to suck on. After some time, the baby presented fever and cervical lymphadenitis. This report emphasizes the importance of serological surveys of toxoplasmosis in nursing infants presenting with fever and lymphadenitis, in view of the possible acquisition of toxoplasmosis in the first months of life.

IgG Avidity in Samples Collected on Filter Paper: Importance of The Early Diagnosis of Congenital Toxoplasmosis / Avidez de IgG em amostras coletadas em papel filtro: Importância no diagnóstico precoce da toxoplasmose congênita

Abstract Objective The purpose of the present study is to standardize and evaluate the use of the immunoglobulin G (IgG) antibody avidity test on blood samples from newborns collected on filter paper to perform the heel test aiming at its implementation in ongoing programs. Methods Blood samples from newborns were collected on filter paper simultaneously with the heel prick test. All samples were subjected to immunoglobulin M IgM and IgG enzyme-linked immunosorbent assays (ELISA). Peripheral blood was collected again in the traditional way and on filter paper from newborns with high IgG levels (33). Three types of techniques were performed, the standard for measuring IgG in serum, adapted for filter paper and the technique of IgG avidity in serum and on filter paper. The results of the avidity test were classified according to the Rahbari protocol. Results Among the 177 samples, 17 were collected in duplicate from the same child, 1 of peripheral blood and 1 on filter paper. In this analysis, 1 (5.88%) of the 17 samples collected in duplicate also exhibited low IgG avidity, suggesting congenital infection. In addition, the results obtained from serum and filter paper were in agreement, that is, 16 (94.12%) samples presented high avidity, with 100% agreement between the results obtained from serum and from filter paper. Conclusion The results of the present study indicate that the avidity test may be another valuable method for the diagnosis of congenital toxoplasmosis in newborns.

Resumo Objetivo O objetivo do presente estudo é padronizar e avaliar a utilização do teste de avidez de anticorpos imunoglobulina G (IgG) em amostras de sangue de recémnascidos (RNs) coletadas em papel filtro para a realização do teste do pezinho visando a implementação nos programas já vigentes. Métodos Foram coletadas amostras de sangue de recém-nascidos em papel filtro simultaneamente ao teste do pezinho. Em todas as amostras, foram realizados os testes imunoenzimáticos (ELISA) imunoglobulina M (IgM) e IgG. Dos RNs que apresentaram altos índices de IgG (33), foi novamente coletado sangue periférico da forma tradicional e em papel filtro. Foram realizadas técnicas padrão para a dosagem de IgG em soro, adaptadas para papel filtro, e a técnica de avidez de IgG em soro e em papel filtro. Os valores obtidos para o teste de avidez foram classificados de acordo com o protocolo de Rahbari. Resultados Dentre as 177 recoletas, em 17 amostras foi realizada a coleta simultânea de sangue periférico e papel filtro da mesma criança. Nesta análise, 1 (5,88%) das 17 amostras coletadas em duplicata obteve também baixa avidez de IgG, sugerindo infecção congênita da criança, e houve concordância entre os resultados obtidos em soro e em papel filtro: 16 (94,12%) das amostras apresentaram alta avidez, com concordância de 100% entre os resultados obtidos em soro e em papel filtro. Conclusão Os dados do presente trabalho evidenciam que o teste de avidez poderá ser mais um método valioso a ser utilizado no diagnóstico da toxoplasmose congênita em RNs.

[Prevalence of toxoplasmosis in pregnant women in two reference centers in a city in Northeast Brazil]. / Prevalência de toxoplasmose em gestantes atendidas em dois centros de referência em uma cidade do Nordeste, Brasil.

PURPOSE: To determine the prevalence of toxoplasmosis and to identify the main factors associated with seroreactivity in pregnant women cared for at two reference centers in a city in Northeast Brazil. METHODS: A cross-sectional study was conducted on 561 pregnant women at two high-risk prenatal reference centers in a city in Northeast Brazil. All women were interviewed using an epidemiological questionnaire and had their blood samples collected for the following serological tests: anti-Toxoplasma gondii IgG and IgM (ELISA), IgG avidity test, and polymerase chain reaction (PCR). Statistical analysis was carried out using SPSS version 18.0 for Windows, calculating odds ratio, confidence interval of 95% and with the level of significance set at 5%. RESULTS: Seroreactivity for toxoplasmosis was detected in 437 women (77.0%), susceptibility in 124 (22.1%) and active infection in 5 (0.9%). There was no significant association between seroreactivity for toxoplasmosis and age, location, income, education, availability of sewage, number of pregnancies or gestational age. The variables significantly associated (p≤0.05) with seroreactivity were multiparity (p=0.03) and living with stray dogs (p=0.01). CONCLUSIONS: This study identified high seroreactivity for toxoplasmosis among patients seen during prenatal care, as well as factors associated with seroreactivity. Appropriate guidelines about primary preventive measures should be emphasized and quarterly serological monitoring is recommended for pregnant women in this city and elsewhere in the Northeast of Brazil.

Occurrence of parasites in salads in restaurants in Aparecida de Goiânia, Goiás, Brazil

Parasitoses are the most widespread diseases in the world. They are transmited via contaminated water or food. Considering that the daily consumption of vegetables is estimated at 142g per person, the purpose of this study was to evaluate the occurrence of parasites in salads available for consumption in restaurants in Aparecida de Goiânia, Goiás State. Salad samples were collected from the restaurants and parasitological analysis was performed using the Willis, Hoffman, Faust and Ziehl Neelsen techniques as well as cultures for the isolation of free-living amoebae. 51 samples were analyzed, 16 (31.4%) were positive. The parasites detected were: Acanthamoeba spp. in 12 (23.5%); free-living larvae, Schistosoma mansoni and Entamoeba coli in 1 (2.0%); Endolimax nana in 2 (3.9%). The PCR technique determined that 17.6% of the samples presented Toxoplasma gondii DNA. These techniques evidenced that the salad samples presented parasite contamination not only in the restaurants with the lowest price per Kg, but also in the most expensive ones. Therefore, in addition to effective sanitary surveillance, prophylactic measures are necessary regarding suppliers, handlers and restaurant owners to prevent the spread of these and other parasites.

Risk factors for Toxoplasma gondii infection in women of childbearing age.

OBJECTIVES: Determine the risk factors involved in toxoplasmosis transmission and determine whether pregnancy is a risk factor for toxoplasmosis infection. STUDY DESIGN: Cross-sectional study carried out on 2,242 women at childbearing age. An indirect immunofluorescence reaction was used to identify immunity to Toxoplasma gondii. Previous gestations were also analyzed as a possible risk factor. The results were analyzed by chi2 and OR tests, and by variance analysis. The sample was statistically balanced according to social-economic risk factors. RESULTS: Previously pregnant women were 1.74 times more frequently infected with toxoplasmosis, regardless of environmental conditions. Pregnant women living under unfavorable environmental conditions had an approximately two times increased risk of being infected for each risk factor (contact with host animals, presence of vehicles of oocyst transmission). Previous pregnancy was the risk factor that had the strongest influence on acquiring toxoplasmosis (variance analysis and statistical balancing). DISCUSSION: The prevalence of this zoonosis is high in Goiânia-GO, Brazil (65.8%). Inadequate environmental sanitation was not significantly correlated with toxoplasmosis infection, except when associated with previous pregnancy, showing that the fundamental cause for infection is not environmental. CONCLUSION: The finding that pregnancy makes women more vulnerable to this protozoan, makes it important to implement prophylactic control of at-risk pregnant women.

Epidemiological factors associated with seropositivity for toxoplasmosis in pregnant women from Gurupi, State of Tocantins, Brazil.

INTRODUCTION: Knowledge of the prevalence and risk factors for Toxoplasma gondii dissemination among pregnant women is relevant because the parasite can be spread from mother to infant. The objective of this study was to assess the epidemiology and risk factors of toxoplasmosis in pregnant women from Gurupi, State of Tocantins, Brazil, from February 2012 to June 2013. METHODS: The study population included 487 pregnant women. Sociodemographic, dietary and cultural data were collected using a standardized and validated form. Peripheral blood was collected for serologic testing using the ELISA test (IgM/IgG antibodies). The data were analyzed by comparing seropositivity with risk factors using crude and adjusted odds ratios. RESULTS: The prevalence rate for IgG and IgM antibodies was 68.7% and 5.7%, respectively. Sociodemographic characteristics associated with toxoplasmosis risk included the following: education level ≤ 8 years (OR: 6.612; CI: 1.450-30.144), age ≥ 30 years (OR: 5.273; CI: 1.166-23.844), working outside the home (OR: 1.604; CI: 1.015-2.536), and family income of two minimum wages or lower (OR: 2.700; CI: 1.891-8.182). Regarding dietary habits, there was a significant association of seropositivity with meat intake (OR: 1.78; CI: 1.149-4.080), cutting vegetables without washing the cutting board beforehand (OR: 2.051; CI: 1.165-3.614), frequent intake of vegetables (OR: 2.051; CI: 1.368-3.006) and in natura milk intake (OR: 2.422; CI: 1.014-5.785). CONCLUSIONS: The high prevalence rates of toxoplasmosis in Gurupi are related to age, raw meat and in natura milk intake, as well as education level, working outside the home, and poor hygienic habits during meal preparation.

Validação do teste sorológico para toxoplasmose em papel filtro / Validation of serological test for toxoplasmosis in filter paper

Objetivo: Validar o uso do teste sorológico para toxoplasmose em papel filtro em amostras de sangue de gestantes do município de Goiânia e região metropolitana. Métodos: Trata-se de um estudo prospectivo, onde foram coletadas 1.006 amostras de sangue em gestantes no município de Goiânia e região metropolitana. Todas as gestantes que concordaram participar da pesquisa assinaram um Termo de Consentimento Livre e Esclarecido. A avaliação do perfil sorológico foi realizada pela técnica de ELISA (ensaio imunoenzimático), com amostras de soro e papel filtro, para pesquisa de anticorpos anti-Toxoplasma gondii das classes IgM e IgG, sendo que o kit utilizado não é padronizado para a pesquisa de anticorpos IgM no papel filtro. A análise estatística foi processada no banco de dados do programa EpiInfo® versão 3.2.1, que avaliou a prevalência de soropositividade, frequência de positividade no soro, frequência de positividade no papel filtro, sensibilidade, especificidade, valor preditivo positivo, valor preditivo negativo e índice kappa. Resultados: Na análise realizada no soro obtiveram-se 421 amostras positivas para anticorpos da classe IgG e três positivas para anticorpos da classe IgM. Em papel filtro obtiveram-se 443 positivas para anticorpos da classe IgG e uma positiva para anticorpos da classe IgM. Conclusão: O estudo permitiu identificar a semelhança de amostras positivas tanto com o uso do soro como no papel filtro para pesquisas de anticorpos da classe IgG, porém, para a pesquisa de anticorpos da classe IgM, o uso do soro apresentou maior sensibilidade quando comparado ao exame realizado com o papel filtro.

Objective: To validate the use of the serological test for toxoplasmosis in filter paper in blood samples from pregnant women from the city of Goiânia and the metropolitan region. Methods: This is a prospective study in which 1,006 blood samples were collected in pregnant women in the city of Goiânia and in the metropolitan region. All pregnant women who agreed to participate in the study signed a Free and Informed Consent Form. Serological profile evaluation was performed using ELISA (immunoenzymatic assay), with serum and filter paper samples, to investigate anti-Toxoplasma gondii antibodies of the IgM and IgG classes, and the kit used is not standardized for the research of IgM antibodies on the filter paper. Statistical analysis was performed in the database of the EpiInfo® version 3.2.1 program, which evaluated the prevalence of seropositivity, frequency of serum positivity, filter paper positivity frequency, sensitivity, specificity, positive predictive value, negative predictive value and index the kappa. Results: In the serum analysis, 421 samples were positive for antibodies of the IgG class and three were positive for antibodies of the IgM class. In the filter paper yielded 443 positive for antibodies of the IgG class and one positive for IgM class antibodies. Conclusion: The study allowed to identify the similarity of positive samples both with the use of serum and in the filter paper for IgG class antibodies. However, for serum IgM antibodies, the use of serum was more sensitive when compared to serum IgG. taken with the filter paper

Plasmodium falciparum malarial parasites from Brazil lack artemisinin resistance-associated mutations in the kelch13 gene

Abstract INTRODUCTION Mutations in the propeller domain of the Plasmodium falciparum kelch13 (k13) gene are associated with artemisinin resistance. METHODS: We developed a PCR protocol to sequence the pfk13 gene and determined its sequence in a batch of 50 samples collected from 2003 to 2016 in Brazil. RESULTS: We identified 1 K189T substitution located outside the propeller domain of the PfK13 protein in 36% of samples. CONCLUSIONS: Although the sample size is relatively small, these results suggest that P. falciparum artemisinin-resistant mutants do not exist in Brazil, thereby supporting the continuation of current treatment programs based on artemisinin-based combination therapy.

Analysis of the accuracy of different laboratory methods for the diagnosis of intestinal parasites from stray and domiciled cats (Felis catus domesticus) in Goiânia, Goiás, Brazil / Análise da acurácia de diferentes técnicas parasitológicas no diagnóstico de parasitos entéricos de gatos (Felis catus domesticus) errantes e domiciliados em Goiânia, Goiás, Brasil

Abstract Cats are carriers of zoonotic agents to humans, including intestinal parasites. The purpose of this study was to analyze the accuracy of different laboratory methods for the diagnosis of intestinal parasites. Fecal samples were processed by the Willis, Sheather, Faust and Hoffman-Janer-Pons-Lutz (HJPL) methods. Accuracy analysis was performed determining the sensitivity, specificity, positive and negative predictive value and Kappa. A total of 149 fecal samples were collected, 65 from stray cats and 84 from domiciled cats. The prevalence of intestinal parasites in stray cats was 60% while in domiciled cats it was 17%. In the analysis of accuracy, the techniques that showed the greatest accuracy for Ancylostomids were Willis and Faust, for Cystoisospora spp. Sheather with Faust or HPJL, and Toxoplasma gondii/Hammondia hammondi the association between Willis and Faust. Therefore, for a reliable evaluation of the prevalence of intestinal parasites, at least two different techniques should be used in parasitological exams of feces.

Resumo Gatos são veiculadores de agentes zoonóticos para o homem, entre eles os parasitos intestinais. O objetivo foi analisar a acurácia de diferentes métodos laboratoriais para o diagnóstico de parasitos entéricos. As amostras de fezes foram processadas pelos métodos de Willis, Sheather, Faust e Hoffman-Janer-Pons-Lutz (HJPL). A análise de acurácia foi realizada determinando a sensibilidade, especificidade, valor preditivo positivo e negativo e índice Kappa. Foram coletadas 65 amostras de fezes de gatos errantes e 84 amostras de gatos domiciliados, total de 149 amostras. A prevalência de parasitos entéricos nos errantes foi 60% e nos domiciliados 17%. Na análise de acurácia as técnicas que associadas apresentaram maior acurácia para Ancilostomídeos foram Willis e Faust, para Cystoisospora spp. Sheather com Faust ou HPJL, e para Toxoplasma gondii/Hammondia hammondi a associação entre Willis e Faust. Portanto, para uma avaliação fidedigna da prevalência de parasitos entéricos, deve-se usar no mínimo duas técnicas diferentes ao realizar os exames parasitológicos de fezes.

Epidemiological factors associated with Toxoplasma gondii infection in postpartum women treated in the public healthcare system of Goiânia, State of Goiás, Brazil

Abstract INTRODUCTION: Knowledge of the epidemiological profile and risk factors for Toxoplasma gondii infection among postpartum women is a relevant issue, because this protozoan can be vertically transmitted to the developing fetus, which can cause severe and debilitating disease. The aim of this study was to assess the risk factors associated with T. gondii infection in postpartum women in Goiânia, GO, Brazil. METHODS: This cross-sectional study comprised 229 postpartum women, among whom 204 were chronically infected (IgG+/IgM-), and 25 were seronegative (IgG-/IgM-; control group). All the patients were asked to complete a form to provide sociodemographic, clinical, dietary, and cultural information. The data were analyzed to compare seropositivity and risk factors based on the odds ratio (OR) thereof. RESULTS: The sociodemographic characteristics associated with the risk for toxoplasmosis were: education ≤ 8 years [OR: 2.521, confidence interval (CI): 1.01-6.301, p=0.049], and age ≥ 30 years (OR: 4.090; CI: 1.180-14.112, p=0.023). Clinical and behavioral characteristics related to eating raw and undercooked meat, were not found to be risk factors associated with a positive test for toxoplasmosis. CONCLUSIONS: Our findings concur with the results of other studies conducted in Brazil and abroad, where variables such as low levels of schooling, and advanced age (≥ 30 years) are major risk factors for pregnant women to become infected with T. gondii.

The first Acanthamoeba keratitis case in the Midwest region of Brazil: diagnosis, genotyping of the parasite and disease outcome

Abstract We report an Acanthamoeba keratitis case associated with the use of contact lens in a 28-year-old female from Brasília, Brazil. Samples from corneal scraping and contact lens case were used for culture establishment, PCR amplification, and partial sequencing (fragments of ~400kb) of small subunit rDNA; both culture and PCR were positive. The sequence analyses of the cornea and of isolates from the contact lens case showed similarity with the T4 genotype. To the best of our knowledge, this is the first report of T4 Acanthamoeba keratitis case from the Midwest region of Brazil.