Emergence of Thymidine-Dependent Staphylococcus aureus Small-Colony Variants in Cystic Fibrosis Patients in Southern Brazil.

We characterized Staphylococcus aureus small-colony variant (SCV) strains isolated from cystic fibrosis (CF) patients in southern Brazil. Smaller colonies of S. aureus were isolated from respiratory samples collected consecutively from 225 CF patients from July 2013 to November 2016. Two phenotypic methods-the auxotrophic classification and a modified method of antimicrobial susceptibility testing-were employed. PCR was conducted to detect the mecA, ermA, ermB, ermC, msrA, and msrB resistance genes. Furthermore, DNA sequencing was performed to determine the mutations in the thyA gene, and multilocus sequence typing was used to identify the genetic relatedness. S. aureus strains were isolated from 186 patients (82%); suggestive colonies of SCVs were obtained in 16 patients (8.6%). The clones CC1 (ST1, ST188, and ST2383), CC5 (ST5 and ST221), and ST398 were identified. Among SCVs, antimicrobial susceptibility testing showed that 77.7% of the isolates were resistant to multiple drugs, and all of them were susceptible to vancomycin. mecA (2), ermA (1), ermB (1), ermC (3), and msrB (18) were distributed among the isolates. Phenotypically thymidine-dependent isolates had different mutations in the thyA gene, and frameshift mutations were frequently observed. Of note, revertants showed nonconservative or conservative missense mutations. SCVs are rarely identified in routine laboratory tests. IMPORTANCE Similar findings have not yet been reported in Brazil, emphasizing the importance of monitoring small-colony variants (SCVs). Altogether, our results highlight the need to improve detection methods and review antimicrobial therapy protocols in cystic fibrosis (CF) patients.

Chemical Composition and In Vitro Antimicrobial Activity of the Essential Oil Obtained from Eugenia pyriformis Cambess. (Myrtaceae)

Abstract Our study aimed to evaluate the chemical composition of the essential oil from the leaves of Eugenia pyriformis Cambess., belonging to the Myrtaceae family and native to the Brazilian Atlantic forest. The volatile compounds in the essential oil were extracted by hydrodistillation and analyzed using GC-MS; 36 compounds accounted for 78.80% of the total oil content. The major compounds were β-caryophyllene, bicyclogermacrene, globulol, and (δ-cadinene. We evaluated their antimicrobial potential of the essential oil and toxicity to Artemia salina. The antimicrobial activity of the essential oil was evaluated against 12 microorganisms using the broth microdilution method. Our results showed moderate inhibition of Staphylococcus aureus and methicillin-resistant S. aureus (MIC, 250 and 125 μg.mL-1, respectively) and toxicity to A. salina (LC50, 125.64 μg.mL-1). Our results establish the biological activity of the essential oil obtained from E. pyriformis.

Erros inatos de imunidade: tempo de diagnóstico e episódios infecciosos em pacientes ambulatoriais / Inborn errors of immunity: time to diagnosis and infections in outpatients

Introdução: Os erros inatos de imunidade (EII) são distúrbios que ocasionam danos no desenvolvimento e/ou função do sistema imunológico. O diagnóstico muitas vezes não é realizado de imediato devido ao pouco conhecimento sobre as doenças, que leva a complicações graves e diminui a sobrevida e qualidade de vida desses pacientes. O objetivo desse estudo foi avaliar o tempo para o diagnóstico e as ocorrências infecciosas que acometeram pacientes com EII no decorrer de sua vida até o momento do diagnóstico. Método: Foi realizado um estudo transversal, retrospectivo, em pacientes atendidos pelo serviço de Alergia, Imunologia e Pneumologia do Complexo Hospital de Clínicas da Universidade Federal do Paraná (CHC-UFPR), no período de junho de 1993 a março de 2019. Foram excluídos pacientes sem história prévia ao diagnóstico e com diagnóstico não confirmado de EII ou indefinido. Resultados: Dos 57 pacientes incluídos no estudo, a maioria (n = 34) era do sexo masculino. A idade ao diagnóstico variou de 2 até 38 anos, sendo a média 9 anos. Dentre as imunodeficiências, 43 (75,4%) tinham deficiência de anticorpos, 10 (17,5%) deficiência combinada, 3 (5,3%) deficiência de fagócitos e 1 (1,8%) deficiência de complemento. Em relação às infecções, os pacientes apresentaram mais de um episódio infeccioso, e também sofreram acometimento em mais de um sítio anatômico. As infecções mais frequentes foram as do trato respiratório inferior (80,7%), seguido das infecções do trato respiratório superior (50,9%). Foi encontrado um atraso médio de diagnóstico de 66,1 meses, sendo que 10,5% dos pacientes foram a óbito. Conclusão: Apesar de já serem bem caracterizados, os EII ainda possuem diagnóstico tardio, levando os pacientes a complicações graves, e até à morte.

Introduction: Inborn errors of immunity (IEIs) cause damage in the development and/or function of the immune system. The diagnosis is often not done immediately because of lack of knowledge about the disorders, leading to serious complications and decreasing the survival and quality of life of these patients. The aim of this study was to evaluate time to diagnosis and occurrence of infections that affected patients with IEI throughout their life-span until the diagnosis. Methods: A retrospective crosssectional study was performed in patients treated at the Division of Allergy, Immunology and Pulmonology of the Complexo Hospital de Clínicas at Universidade Federal do Paraná, from June 1993 to March 2019. Patients with no history prior to diagnosis and those with unconfirmed diagnosis of IIE or undefined diagnosis were excluded from the study. Results: Of the 57 patients included in the study, most were male (n = 34). The mean age at the time of diagnosis was 9 years, ranging from 2 to 38 years. Among the immunodeficiencies, 43 (75.4%) patients had antibody deficiency disorder, 10 (17.5%) had combined immunodeficiency, 3 (5.3%) had phagocyte deficiency and 1 (1.8%) had complement disorder. Regarding infections, patients had more than one infectious episode and were affected in more than one anatomical site. The most frequent infections were those of the lower respiratory tract (80.7%), followed by upper respiratory tract infections (50.9%). A mean diagnosis delay of 66.1 months was found, and 10% of the patients died. Conclusion: Despite being well characterized, IEIs still have late diagnosis, leading patients to serious complications and even death.

Comparison of Cerebrospinal Fluid Biomarkers for Differential Diagnosis of Acute Bacterial and Viral Meningitis with Atypical Cerebrospinal Fluid Characteristics.

OBJECTIVE: Several cerebrospinal fluid (CSF) biomarkers are used to distinguish between acute bacterial meningitis (BM) and viral meningitis (VM). We compared the ability of lactate and glucose (GL) in CSF and the CSF/blood GL ratio to distinguish between acute BM and VM with typical and atypical CSF characteristics. METHODS: Three hundred and twenty-four CSF reports were included, which were distributed as the acute BM, VM, and normal control groups (n = 63, 139, and 122, respectively). RESULTS: Lactate level in the CSF of acute BM group was 4-fold higher than that in the acute VM and control groups (p < 0.0001). CSF lactate presented higher specificity (92%) and negative predictive value (94%) compared to CSF GL and CSF/blood GL ratio in distinguishing acute BM and VM. Definitive acute BM or VM with atypical CSF cell characteristics was observed in 23.2 and 21.6% of samples, respectively, and these groups showed reduced performance of characteristics of all CSF biomarkers. CSF lactate showed better operational characteristics than those of CSF GL and CSF/blood GL ratio, presenting the highest positive likelihood ratio, and thus aided in the differential diagnosis of VM with atypical CSF. CONCLUSION: The CSF lactate assay can be routinely used in laboratories as a rapid, automated, and easy method that is independent of lactate blood levels.

Diagnostic importance of eosinophilic meningitis in HIV-positive and HIV-negative patients.

The presence of eosinophils in the cerebrospinal fluid (CSF) should always be considered abnormal. This study aimed to evaluate the causes of eosinophils in the CSF of patients who are HIV positive and HIV negative. This is the first study of eosinophils in the CSF of patients who are HIV-positive. This was a retrospective study of CSF reports from 1996 to 2005, patients were selected based on the presence of eosinophils in the CSF. We analyzed 20,008 CSF reports; eosinophils were present in 5%. The median and interquartile range (IQR) of eosinophils was 2% (1%, 4%). Eosinophilic meningitis (CSF eosinophils ≥ 10%) was present in 12% of the samples. The main etiologies were infectious diseases as follows: neurocysticercosis, Cryptococcus sp. meningitis, and acute bacterial meningitis. In HIV-positive cases, all causes were by infectious disease, the main pathogen being Cryptococcus sp. The probability of neurocysticercosis in a patient from an endemic region who is HIV-negative and has CSF eosinophils more than 10% was five times higher compared to a person without eosinophilic meningitis. There was a weak positive correlation between CSF eosinophils and increased serum eosinophils. Among the HIV-negative cases, the most frequent non-infectious causes were cerebrovascular syndromes, of these hemorrhage (91.5%). In the HIV-positive group, there were no cases of non-infectious cerebral disease. CSF eosinophils are suggestive of disease. The causes must be investigated, considering the most prevalent infectious diseases in the region.

Ralstonia mannitolilytica bacteremia in a neonatal intensive care unit.

Ralstonia mannitolilytica, a Gram-negative bacterium, is rarely isolated in clinical laboratories. It has been associated with outbreaks due to its ability to survive in liquid media and hospital devices. We describe three cases of bacteremia caused by R. mannitolilytica in a neonatal intensive care unit in Curitiba, Southern Brazil. All isolates presented the same PFGE profile. The common source of infection was undetected in surveillance cultures for the outbreak survey. All patients received antimicrobial treatment and were discharged from the maternity. Due to the characteristics of the microorganism, clinicians and microbiologists should pay attention to the emergence of Ralstonia spp. infections.

Ralstonia mannitolilytica bacteremia in a neonatal intensive care unit

Abstract Ralstonia mannitolilytica, a Gram-negative bacterium, is rarely isolated in clinical laboratories. It has been associated with outbreaks due to its ability to survive in liquid media and hospital devices. We describe three cases of bacteremia caused by R. mannitolilytica in a neonatal intensive care unit in Curitiba, Southern Brazil. All isolates presented the same PFGE profile. The common source of infection was undetected in surveillance cultures for the outbreak survey. All patients received antimicrobial treatment and were discharged from the maternity. Due to the characteristics of the microorganism, clinicians and microbiologists should pay attention to the emergence of Ralstonia spp. infections.

Is it possible to perform bacterial identification and antimicrobial susceptibility testing with a positive blood culture bottle for quick diagnosis of bloodstream infections?

INTRODUCTION: Bloodstream infections can be fatal, and timely identification of the etiologic agent is important for treatment. METHODOLOGY: An alternative method, consisting of direct identification and susceptibility testing of blood culture bottles using the automated VITEK 2® system, was assessed. RESULTS: All 37 of the Gram-negative bacilli (GNB) identifications and 57.1% of the 28 Gram-positive cocci (GPC) identifications matched those obtained with standard methods. In susceptibility testing, the agreement was greater than 90%. CONCLUSIONS: This alternative methodology may assist in the early identification and susceptibility testing of GNB. Further research is necessary to develop appropriate methods for GPC.

Is it possible to perform bacterial identification and antimicrobial susceptibility testing with a positive blood culture bottle for quick diagnosis of bloodstream infections?

Abstract INTRODUCTION: Bloodstream infections can be fatal, and timely identification of the etiologic agent is important for treatment. METHODOLOGY: An alternative method, consisting of direct identification and susceptibility testing of blood culture bottles using the automated VITEK 2® system, was assessed. RESULTS: All 37 of the Gram-negative bacilli (GNB) identifications and 57.1% of the 28 Gram-positive cocci (GPC) identifications matched those obtained with standard methods. In susceptibility testing, the agreement was greater than 90%. CONCLUSIONS: This alternative methodology may assist in the early identification and susceptibility testing of GNB. Further research is necessary to develop appropriate methods for GPC.

The detection of Bence Jones protein in urine by the heat test helps in diagnosis of multiple myeloma? / A detecção de proteína Bence Jones na urina pelo teste de calor auxilia no diagnóstico de mieloma múltiplo?

ABSTRACT Introduction: Multiple myeloma (MM) is a hematologic malignancy caused by the intense indiscriminate proliferation of plasma cells in the bone marrow. In view of clinical suspicion of MM, clinic laboratory tests and imaging tests should be used, among others. Objectives: Evaluate the laboratory test for protein detection heat method Bence Jones (BJ) used to complementary diagnosis pathology and to characterize the epidemiological profile of patients diagnosed with MM. Material and methods: A retrospective study was conducted from January 2010 to July 2015 of the patients treated at Hospital de Clínicas of Universidade Federal do Paraná (HC/UFPR) in Curitiba, Paraná, Brazil. Results: In the patients analyzed, the average age at diagnosis of MM was 65.6 years, with a minimum percentage of difference between genders [males 52.6% (n = 10) and females 47.4% (n = 9)], predominantly in the white race [84.2% (n = 16)]. Among the patients analyzed, 85.2% (n = 104) had negative BJ exam and 14.8 (n = 18), positive exam; 84.4% (n = 103) had no diagnosis of MM, and 15.6% (n = 19) were diagnosed with the disease. Conclusion: The evaluation results of BJ protein detection by the heat method showed sensitivity of 47.4%, specificity of 91.3%, with positive and negative predictive values of 50% and 90.4%, respectively.

RESUMO Introdução: O mieloma múltiplo (MM) é uma neoplasia maligna hematológica causada pela intensa proliferação indiscriminada de plasmócitos na medula óssea. Diante da suspeita clínica de MM devem ser realizados exames laboratoriais e exames de imagem, entre outros. Objetivos: Avaliar o teste laboratorial de calor para detecção de proteína Bence Jones (BJ), utilizado como diagnóstico complementar da patologia, e caracterizar o perfil epidemiológico dos pacientes diagnosticados com MM. Material e métodos: Foi realizado um estudo retrospectivo de janeiro de 2010 a julho de 2015 dos pacientes atendidos no laboratório do Hospital de Clínicas da Universidade Federal do Paraná (UFPR), Curitiba, Paraná, Brasil. Resultados: Dos pacientes avaliados, a média de idade no momento do diagnóstico de MM foi de 65,6 anos, com um percentual de diferença mínima entre os gêneros masculino [52,6% (n = 10)] e feminino [47,4% (n = 9)] e predomínio na raça branca [84,2% (n = 16)]. Entre os pacientes analisados, 85,2% (n = 104) apresentaram exame de BJ negativo e 14,8 (n = 18), positivo; 84,4% (n = 103) não apresentaram diagnóstico de MM e 15,6% (n = 19) foram diagnosticados com a patologia. Conclusão: Os resultados da avaliação do método de calor de detecção de proteína BJ mostraram sensibilidade de 47,4% e especificidade de 91,3%, com valores preditivos positivo e negativo de 50% e 90,4%, respectivamente.

Microbiological quality of Brazilian artisanal cheese and fermented sausages / Qualidade microbiológica de queijos e salames artesanais brasileiros

Cheeses and sausages are ready-to-eat foods, and the products prepared by artisanal processare susceptible to microbial contamination. Few studies on the quality of foods from different Brazilian regions have been done. In this context, this study aimed at evaluating the microbiological quality of 32 cheese and 13 sausages samples purchased from artisanal food stores or producers markets located in the metropolitan areas of 10 capital cities of Brazilian states. Microbiological analyses were performed to determine the counts of microbial contamination indicators, including Escherichia coli and coagulase-positive staphylococci, as well as for evaluating the presence of Listeria monocytogenes and Salmonella spp. using bacteriological methods.E. coli was detected in 50.0 % of samples, coagulase-positive staphylococci in 34.4 %, and Salmonella spp. in 6.3 % of cheese samples. In fermented sausage samples, coagulase-positive staphylococci were isolated from 23.1 % samples and Salmonella spp. from 7.7 %. According tothe Brazilian Sanitary Legislation, 63.0 % of cheese samples and 23.0 % of artisanal fermentedsausage samples from the metropolitan areas of 10 Brazilian capital cities were unsuitable for consumption, indicating the importance of conducting the close monitoring of these foodsand the application of the effective measures for preventing food-borne outbreaks.

Queijos e salames são alimentos prontos para consumo e quando artesanalmente produzidos são suscetíveis à contaminação microbiana. A importância deste estudo em dez capitais brasileiras deve-se à carência de dados obtidos simultaneamente abrangendo-se diferentes regiões do Brasil. O objetivo do presente trabalho foi de avaliar a qualidade microbiológica de 32 amostras de queijos e 13 de salames artesanais, adquiridos em casas de produtos artesanais ou feiras de produtores nas regiões metropolitanas de dez capitais brasileiras. Análises microbiológicas com as respectivas contagens de indicadores de contaminação microbiana, Escherichia coli e Staphylococcus coagulase positiva, e a pesquisa de Listeria monocytogenes e Salmonella spp. foram realizadas por meio de métodos bacteriológicos. Nas amostras de queijos foram observadas E. coli em 50,0 %, Staphylococcus coagulase positiva em 34,4 % e Salmonella spp. em 6,3 %. Nas amostras de salames foram detectadas Staphylococcus coagulase positiva em 23,1 % e Salmonella spp. em 7,7 %. De acordo com a legislação sanitária brasileira, 63,0 % das amostras de queijos e 23,0 % das amostras de salames artesanais coletadas da Região Metropolitana de dez capitais brasileiras estavam impróprias para o consumo, o que demonstra a importância de realizar monitoramento próximo e efetivo para prevenir surtos de origem alimentar.

Microbiological quality of Brazilian artisanal cheese and fermented sausages / Qualidade microbiológica de queijos e salames artesanais brasileiros

Cheeses and sausages are ready-to-eat foods, and the products prepared by artisanal process are susceptible to microbial contamination. Few studies on the quality of foods from different Brazilian regions have been done. In this context, this study aimed at evaluating the microbiological quality of 32 cheese and 13 sausages samples purchased from artisanal food stores or producers markets located in the metropolitan areas of 10 capital cities of Brazilian states. Microbiological analyses were performed to determine the counts of microbial contamination indicators, including Escherichia coli and coagulase-positive staphylococci, as well as for evaluating the presence of Listeria monocytogenes and Salmonella spp. using bacteriological methods. E. coli was detected in 50.0 % of samples, coagulase-positive staphylococci in 34.4 %, and Salmonella spp. in 6.3 % of cheese samples. In fermented sausage samples, coagulase-positive staphylococci were isolated from 23.1 % samples and Salmonella spp. from 7.7 %. According to the Brazilian Sanitary Legislation, 63.0 % of cheese samples and 23.0 % of artisanal fermented sausage samples from the metropolitan areas of 10 Brazilian capital cities were unsuitable for consumption, indicating the importance of conducting the close monitoring of these foods and the application of the effective measures for preventing food-borne outbreaks.

Queijos e salames são alimentos prontos para consumo e quando artesanalmente produzidos são suscetíveis à contaminação microbiana. A importância deste estudo em dez capitais brasileiras deve-se à carência de dados obtidos simultaneamente abrangendo-se diferentes regiões do Brasil. O objetivo do presente trabalho foi de avaliar a qualidade microbiológica de 32 amostras de queijos e 13 de salames artesanais, adquiridos em casas de produtos artesanais ou feiras de produtores nas regiões metropolitanas de dez capitais brasileiras. Análises microbiológicas com as respectivas contagens de indicadores de contaminação microbiana, Escherichia coli e Staphylococcus coagulase positiva, e a pesquisa de Listeria monocytogenes e Salmonella spp. foram realizadas por meio de métodos bacteriológicos. Nas amostras de queijos foram observadas E. coli em 50,0 %, Staphylococcus coagulase positiva em 34,4 % e Salmonella spp. em 6,3 %. Nas amostras de salames foram detectadas Staphylococcus coagulase positiva em 23,1 % e Salmonella spp. em 7,7 %. De acordo com a legislação sanitária brasileira, 63,0 % das amostras de queijos e 23,0 % das amostras de salames artesanais coletadas da Região Metropolitana de dez capitais brasileiras estavam impróprias para o consumo, o que demonstra a importância de realizar monitoramento próximo e efetivo para prevenir surtos de origem alimentar.

Research on antimicrobial residues activity in urine samples of hospitalized patients / Pesquisa da atividade de resíduos de antimicrobianos em amostras de urina de pacientes hospitalizados

Introduction: Urinary tract infection is quite frequent in a hospital environment, and the urine culture is the gold standard for diagnosis of this disease, because it allows bacterial identification and performing antimicrobial susceptibility testing. Culturenegative urine samples result of patients with strong suspicion of infection may occur due to the activity of antimicrobial residues, which can interfere with bacterial growth in vitro and produce false-negative results. Objective: Verify the occurrence of falsenegative urine cultures due to the presence of antimicrobial residues in samples of patients admitted to the Clinical Hospital of Paraná Federal University. Material and methods: A total of 188 urine samples from hospitalized patients were randomly selected, during the period from July to December 2012. All samples were evaluated on the result of the urine culture, bacteriuria, and research on residues of antimicrobial activity by manual and automated techniques. Results: 44 (23.4%) presented positive urine culture, 121 (64.4%) negative urine culture, and 23 (12.2%) presented growth of many species. In 14 samples, negative urine cultures associated with the presence of bacteria and were positive for the research on antimicrobial residues activity (RARA), were observed. Conclusion: Automated technique showed better performance when compared to manual technique, with sensitivity of 92.8% and 71.4%, respectively. The presence of antimicrobial residues may affect the recovery of bacteria in the urine, producing a false-negative result. .

Introdução: A infecção do trato urinário é bastante frequente em ambiente hospitalar, e a cultura de urina é padrão-ouro para o diagnóstico dessa doença, pois permite a identificação bacteriana e a realização do teste de suscetibilidade aos antimicrobianos. Amostras de urina de pacientes com forte suspeita de infecção que resultam em cultura negativa podem ocorrer devido à atividade de resíduos de antimicrobianos, os quais podem interferir no crescimento bacteriano in vitro e gerar resultados falso negativos. Objetivo: Verificar a ocorrência de culturas de urina falso negativas devido à presença de resíduos de antimicrobianos em amostras de pacientes internados no Hospital de Clínicas da Universidade Federal do Paraná. Material e métodos: Um total de 188 amostras de urina de pacientes internados foi selecionado aleatoriamente, durante o período de julho a dezembro de 2012. Todas as amostras foram avaliadas quanto ao resultado da cultura de urina, da bacteriúria e da pesquisa da atividade de resíduos de antimicrobianos por meio das técnicas manual e automatizada. Resultados: Quarenta e quatro (23,4%) apresentaram cultura de urina positiva, 121 (64,4%), cultura negativa e 23 (12,2%), crescimento de várias espécies. Em 14 amostras foi observada cultura negativa associada à presença de bactérias e pesquisa da atividade de resíduos de antimicrobianos (PRA) positiva. Conclusão: A técnica automatizada apresentou melhor desempenho quando comparada com a técnica manual, apresentando sensibilidade de 92,8% e 71,4%, respectivamente. A presença de resíduos de antimicrobianos pode comprometer a recuperação de bactérias na urina, gerando resultado falso negativo. .

Emergence of extended-spectrum ß-lactamase producing Enterobacter spp. in patients with bacteremia in a tertiary hospital in southern Brazil.

BACKGROUND: Extended-spectrum ß-lactamases (ESBLs) are increasingly prevalent in Enterobacter spp., posing a challenge to the treatment of infections caused by this microorganism. The purpose of this retrospective study was to evaluate the prevalence, risk factors, and clinical outcomes of inpatients with bacteremia caused by ESBL and non ESBL-producing Enterobacter spp. in a tertiary hospital over the period 2004-2008. METHODS: The presence of blaCTX-M, blaTEM, blaSHV, and blaPER genes was detected by polymerase chain reaction (PCR) and nucleotide sequence analysis. Genetic similarity between strains was defined by pulsed-field gel electrophoresis (PFGE). RESULTS: Enterobacter spp. was identified in 205 of 4907 of the patients who had positive blood cultures during hospitalization. Of those cases, 41 (20%) were ESBL-producing Enterobacter spp. Nosocomial pneumonia was the main source of bacteremia caused by ESBL-producing Enterobacter spp. The presence of this microorganism was associated with longer hospital stays. The ESBL genes detected were: CTX-M-2 (23), CTX-M-59 (10), CTX-M-15 (1), SHV-12 (5), and PER-2 (2). While Enterobacter aerogenes strains showed mainly a clonal profile, Enterobacter cloacae strains were polyclonal. CONCLUSION: Although no difference in clinical outcomes was observed between patients with infections by ESBL-producing and non-ESBL-producing strains, the detection of ESBL in Enterobacter spp. resulted in the change of antimicrobials in 75% of cases, having important implications in the decision-making regarding adequate antimicrobial therapy.

Characterization of virulence genes cagA and vacA in Helicobacter Pylori and their prevalence in gastrointestinal disorders

Prevalence of H. pylori infection was determined using cultures of gastric biopsy samples of patients attended at the academic hospital of the Federal University of Paraná, Curitiba, Paraná, Brazil. Molecular methods were used to characterize the cagA and vacA genes from bacterial isolates associated with different diseases presented by patients. Out of a total of 81, forty-two gastric biopsy samples tested were positive for H. pylori, with a prevalence of 51.9 percent. No significant difference was found with regard to the gender (p=0.793) and age (p=0.183) of the patients. Genotype s1m1 vacA gene was found in 67 percent of the cases of peptic ulcer investigated (p=1.0), despite the limited number of patients with this disease (n=3). A correlation between the presence of less virulent strains (s2m2) and reflux esophagitis was found in the majority of the cases (45 percent), but without statistical significance. An association between the prevalence of cagA gene, found in 92 percent of isolates, and peptic ulcer was not observed (p=1.0), suggesting that this gene cannot be considered a specific marker of severity in our environment. The results reinforce the importance of conducting regional studies and the need to characterize H. pylori virulence genes associated with different diseases.

Quantification of cerebrospinal fluid lactic acid in the differential diagnosis between HIV chronic meningitis and opportunistic meningitis.

BACKGROUND: Approximately 40% of HIV infected patients have chronic meningitis at various stages during the infection, 59% are asymptomatic. This is a diagnosis of exclusion and a confounding factor in cerebrospinal fluid (CSF) analysis, any other causes of chronic meningitis by opportunistic or co-infection must be ruled out. The aim of this study was to analyze CSF lactic acid (LA) as an adjuvant biomarker in chronic meningitis due to HIV. METHODS: CSF LA was quantified in 223 CSF samples by the Dimension AR (Dade Behring, Deerfield, IL, USA), distributed into nine groups: 1) HIV positive with an increase in CSF WBCs (n=17); 2) HIV positive with normal CSF (n=20); 3) enterovirus meningitis (n=33); 4) Herpesviridae meningoencephalitis (n=30); 5) fungal meningitis (n=25); 6) tuberculosis (TB) meningitis (n=17); 7) toxoplasmosis (n=18); 8) neurosyphilis (n=6); 9) control group (n=57). RESULTS: CSF LA (median; IQR) was higher in samples with TB meningitis (5.5; 2.9-7.5 mmol/L) and Cryptococcus neoformans meningitis (3.9; 2.7-5.8 mmol/L) compated with samples with HIV chronic meningitis (1.7; 1.4-1.9 mmol/L) and other groups (p ≤ 0.0001). For the diagnosis of HIV chronic meningitis, using a cut-off of 3.5 mmol/L, CSF LA showed high sensitivity and negative predictive value, although low specificity. CONCLUSIONS: CSF LA helps to discriminate between C. neoformans or TB meningitis and HIV chronic meningitis: CSF LA can be included with the methods currently used to identify these specific pathogens, though it does not replace them. It is rapid, inexpensive and easy to perform, and can be used in developing countries.

Characterization of virulence genes cagA and vacA in Helicobacter Pylori and their prevalence in gastrointestinal disorders.

Prevalence of H. pylori infection was determined using cultures of gastric biopsy samples of patients attended at the academic hospital of the Federal University of Paraná, Curitiba, Paraná, Brazil. Molecular methods were used to characterize the cagA and vacA genes from bacterial isolates associated with different diseases presented by patients. Out of a total of 81, forty-two gastric biopsy samples tested were positive for H. pylori, with a prevalence of 51.9%. No significant difference was found with regard to the gender (p=0.793) and age (p=0.183) of the patients. Genotype s1m1 vacA gene was found in 67% of the cases of peptic ulcer investigated (p=1.0), despite the limited number of patients with this disease (n=3). A correlation between the presence of less virulent strains (s2m2) and reflux esophagitis was found in the majority of the cases (45%), but without statistical significance. An association between the prevalence of cagA gene, found in 92% of isolates, and peptic ulcer was not observed (p=1.0), suggesting that this gene cannot be considered a specific marker of severity in our environment. The results reinforce the importance of conducting regional studies and the need to characterize H. pylori virulence genes associated with different diseases.

Phenotypic and genotypic characterization of group B streptococcal isolates in southern Brazil.

One-hundred sixty-eight group B streptococcal (GBS) isolates from a Brazilian hospital were phenotypically and genotypically characterized. Isolates were recovered from human sources from April 2006 to May 2008 and classified as either invasive, noninvasive, or colonizing isolates. Classical methods for serotyping and antibiotic resistance profiling were employed. Clonal groups were also defined by pulsed-field gel electrophoresis (PFGE). Results showed that susceptibility to beta-lactam antimicrobials was predominant among the isolates. Only 4.7% were resistant to erythromycin and clindamycin. The erm(B) gene was widely detected in our GBS isolates, according to our phenotypic results (constitutive macrolide-lincosamide-streptogramin B [cMLSB] resistance phenotype), and the erm(A) gene was also detected in some isolates. MLSB resistance was restricted to strains isolated from patients with noninvasive infections and carriers. Serotype Ia was predominant (38.1%), serotype IV isolates were found at a high frequency (13.1%), and few isolates of serotype III were identified (3%). Pulsed-field gel electrophoresis results revealed a variety of types, reflecting the substantial genetic diversity among GBS strains, although a great number of isolates could be clustered into two major groups with a high degree of genetic relatedness. Three main PFGE clonal groups were found, and isolates sharing the same PFGE type were grouped into different serotypes. Furthermore, in a few cases, isolates from the same patients and possessing the same PFGE type were of different serotypes. These findings could be related to the occurrence of capsular switching by horizontal transfer of capsular genes.

Anti-Helicobacter pylori activity of plant extracts traditionally used for the treatment of gastrointestinal disorders

The antibacterial activity of plant extracts obtained from Bixa orellana L., Chamomilla recutita L., Ilex paraguariensis A. St.-Hil., Malva sylvestris L., Plantago major L. and Rheum rhaponticum L. has been evaluated against two reference strains and eleven clinical isolates of Helicobacter pylori. All the plant species chosen are used in popular Brazilian cuisine and folk medicine in the treatment of gastrointestinal disorders. Initial screening was made by the disk diffusion test and then minimum inhibitory concentration was determined by the agar dilution method. The results presented in this work demonstrated that among the plant preparations analyzed, B. orellana L., C. recutita L., I. paraguariensis A. St.-Hil. and M. sylvestris L. were capable of inhibiting the in vitro growth of H. pylori.