Different virulence genetic context of multidrug-resistant CTX-M- and KPC-producing Klebsiella pneumoniae isolated from cerebrospinal fluid.

Information regarding resistance and virulence traits of meningitis-causing enterobacteria in hospital environment remains scarce. The aim of this study was to characterize virulence and acquired resistance genes of carbapenem-resistant and/or 3rd to 4th generation cephalosporin-resistant Klebsiella pneumoniae isolated from the cerebrospinal fluid of inpatients. Antimicrobial susceptibility testing was performed by disk diffusion. The string test was performed to identify hypermucoviscous phenotype. Galleria mellonella infection model was used to evaluate the virulence profile of the isolates. Screening for virulence determinants and acquired resistance genes were performed by PCR. The blaCTX-M and/or blaKPC and/or rmtG were detected in all the isolates. Genetic virulence determinants, including mrkD, entB, iroD, fecIRA, uge, wabG, fimH, ureA, ybtS, and clb were detected in the majority of multidrug-resistant K. pneumoniae isolates. One isolate presented hypermucoviscous phenotype, and several isolates showed enhanced virulence in G. mellonella infection model. The combination of the virulence genes found here seems to support not only the known virulence genetic context among nosocomial infections-causing K. pneumoniae but also the role that clb and ybtS may play in K. pneumoniae virulence.

Isolamento de Campylobacter sp na região nordeste do Estado de São Paulo – Brasil / Isolation of Campylobacter sp in northeast region of São Paulo State – Brazil

As espécies de Campylobacter podem ser patogênicas ou comensais do trato gastrointestinal de animais domésticos e silvestres, sendo dispersas no ambiente contaminando água, pastos e plantações. São isoladas a partir do meio ambiente e animais, especialmente mamíferos, pássaros e aves domésticas. As espécies termófilas, principalmente C. jejuni e C. coli são reconhecidas mundialmente como agentes etiológicos de diarreia aguda em seres humanos, bem como infecção extraintestinal. Este estudo teve como objetivo avaliar a ocorrência de campilobacteriose nas análises realizadas pelo Centro de Laboratório Regional de Ribeirão Preto VI, Instituto Adolfo Lutz (CLR-RP-VI/IAL), provenientes da Rede Regional de Assistência à Saúde (RRAS-13), São Paulo - Brasil, durante o período de 1987 a 2013. Realizou-se o isolamento e a identificação de Campylobacter sp de acordo com metodologia tradicional. Avaliaram-se 2.534 amostras de fezes das quais 279 (11,0%) culturas foram positivas para Campylobacter sp: 63,1% C. jejuni; 3,9% C. coli; 0,4% C. laridis; 32,6% Campylobacter spp. Em 44 (15,8%) amostras ocorreu associação de Campylobacter sp com outras bactérias patogênicas. Isolou-se três casos em infeccções extraintestinais. É necessária vigilância constante para reduzir a ocorrência de campilobacterioses na região estudada, especialmente C. jejuni...

Antimicrobial resistance in neisseria gonorrhoeae isolates from ribeirão preto, São Paulo, Brazil

Gonorrhea is sexually transmitted, with a high incidence worldwide. Occurrence of resistance and difficulties in treatment is often reported. Penicillin is not used anymore, and quinolones or cephalosporins are the remaining therapeutic options. However, there are resistance reports to these drugs as well. Objective: to evaluate the occurrence of resistance to penicillin, tetracycline, ofloxacin, ciprofloxacin, azithromycin and ceftriaxone in Neisseria gonorrhoeae (gonococcus). Methods: ninety-three endocervical and urethral secretion samples, suspected of gonorrhea, were cultured from September 2008 to May 2012. Samples were collected at the STD/Aids Reference Center and processed at Instituto Adolfo Lutz in Ribeirão Preto, SP, Brazil. Antimicrobial susceptibility tests were performed by the E-test (Oxoid). Beta-lactamase was determined by the cefinase disk method (BD BBL). The susceptibility study included a gonococcus isolated from a case of conjunctivitis. Results: gonococcus was isolated in 41.9% (35) of the cases in the study. Male patients were predominant in 92.3% of samples, with ages ranging from 14 to 62 years, and the conjunctivitis isolate was recuperated from a 1 month old patient. Isolates were resistant to penicillin (44.4%); tetracycline (55.5%); ofloxacin (36.1%) and ciprofloxacin (36.1%). All isolates were susceptible to ceftriaxone, and 80.6% were susceptible to azithromycin. The beta-lactamase test was positive for 31.0% of isolates. Conclusion: in vitro results showed that tetracycline was less effective, and ceftriaxone the most effective antibiotic against gonococcus. The resistance to different drugs limits the options of gonococcus effective treatment.

Phenotypic and genotypic characterization of Rhodococcus equi isolated from sputum

INTRODUCTION: Rhodococcus equi is an opportunistic pathogen, causing rhodococcosis, a condition that can be confused with tuberculosis. Often, without identifying M. tuberculosis, physicians initiate empiric treatment for tuberculosis. R. equi and M. tuberculosis have different susceptibility to drugs. Identification of R. equi is based on a variety of phenotypic, chromatographic, and genotypic characteristics. OBJECTIVE: This study aimed to characterize bacterial isolates from sputum samples suggestive of R. equi. METHODS: The phenotypic identification included biochemical assays; thin-layer chromatography (TLC) and polymerase chain reaction (PCR) were used for genotypic identification. RESULTS: Among 78 Gram-positive and partially acid-fast bacilli isolated from the sputum of tuberculosis-suspected patients, 51 were phenotypically and genotypically characterized as R. equi based on literature data. Mycolic acid analysis showed that all suspected R. equi had compounds with a retention factor (Rf) between 0.4-0.5. Genotypic characterization indicated the presence of the choE gene 959 bp fragments in 51 isolates CAMP test positive. Twenty-two CAMP test negative isolates were negative for the choE gene. Five isolates presumptively identified as R. equi, CAMP test positive, were choE gene negative, and probably belonged to other bacterial species. CONCLUSIONS: The phenotypic and molecular techniques used constitute a good methodological tool to identify R. equi.

Phenotypic and genotypic characterization of Rhodococcus equi isolated from sputum.

INTRODUCTION: Rhodococcus equi is an opportunistic pathogen, causing rhodococcosis, a condition that can be confused with tuberculosis. Often, without identifying M. tuberculosis, physicians initiate empiric treatment for tuberculosis. R. equi and M. tuberculosis have different susceptibility to drugs. Identification of R. equi is based on a variety of phenotypic, chromatographic, and genotypic characteristics. OBJECTIVE: This study aimed to characterize bacterial isolates from sputum samples suggestive of R. equi. METHODS: The phenotypic identification included biochemical assays; thin-layer chromatography (TLC) and polymerase chain reaction (PCR) were used for genotypic identification. RESULTS: Among 78 Gram-positive and partially acid-fast bacilli isolated from the sputum of tuberculosis-suspected patients, 51 were phenotypically and genotypically characterized as R. equi based on literature data. Mycolic acid analysis showed that all suspected R. equi had compounds with a retention factor (R(f)) between 0.4-0.5. Genotypic characterization indicated the presence of the choE gene 959bp fragments in 51 isolates CAMP test positive. Twenty-two CAMP test negative isolates were negative for the choE gene. Five isolates presumptively identified as R. equi, CAMP test positive, were choE gene negative, and probably belonged to other bacterial species. CONCLUSIONS: The phenotypic and molecular techniques used constitute a good methodological tool to identify R. equi.

Investigação de meningite por Neisseria meningitidis, Streptococcus pneumoniae e Haemophilus influenzae na região de Ribeirão Preto, SP, Brasil, utilizando métodos laboratoriais convencionais / Investigation of meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae in Ribeirão Preto, SP, Brazil, using conventional laboratory methods

A meningite é uma doença que acomete as populações de diferentes partesdo mundo, sendo caracterizada como um importante problema de saúdepública. O objetivo deste estudo foi avaliar a positividade para Neisseriameningitidis, Streptococcus pneumoniae e Haemophilus influenzaeutilizando diferentes métodos convencionais de diagnóstico laboratorial. O estudo incluiu os casos suspeitos de meningite bacteriana diagnosticados no Instituto Adolfo Lutz de Ribeirão Preto, ocorridos no período de janeiro de 2007 a dezembro de 2009. Foram analisadas 1.722 amostras utilizando, individualmente ou associados, os métodos de bacterioscopia, cultura delíquido cefalorraquidiano (LCR), hemocultura e/ou reação decontraimunoeletroforese. Entre as amostras analisadas, 392 (22,76%)apresentaram positividade para algum agente bacteriano em pelo menosum dos métodos utilizados. Dessas, 17,35% foram positivas para N.meningitidis, 15,82% para S. pneumoniae e 2,04% para H. influenzae. O fenótipo C: 23: P1.14-6 de N. meningitidis foi o mais prevalente, seguido do B: 4,7: P1.19,15. Os sorotipos 14, 3, 12F, 23F, 19F e 6B de S. pneumoniae foram os mais encontrados. O estudo concluiu que todos os métodos de diagnóstico laboratorial utilizados são complementares e nenhum delesdeve ser negligenciado. A cultura continua sendo considerada o padrãoouro para o diagnóstico dameningite bacteriana

Rhodococcus equi isolation from sputum of patients with suspected tuberculosis.

Rhodococcus equi has emerged as an opportunistic pathogen associated with pulmonary, invasive or systemic infections in immunocompromised patients. We report the identification of 51 R. equi isolates found in sputum samples of 546 individuals suspected to have pulmonary tuberculosis in two Public Health Hospital Units in Brazil. The epidemiology of R. equi infection as well as the phenotypic identification and drug susceptibility profile of isolates are described in this paper.

Rhodococcus equi isolation from sputum of patients with suspected tuberculosis

Rhodococcus equi has emerged as an opportunistic pathogen associated with pulmonary, invasive or systemic infections in immunocompromised patients. We report the identification of 51 R. equi isolates found in sputum samples of 546 individuals suspected to have pulmonary tuberculosis in two Public Health Hospital Units in Brazil. The epidemiology of R. equi infection as well as the phenotypic identification and drug susceptibility profile of isolates are described in this paper.

Nasopharyngeal colonization by Haemophilus influenzae in children attending day-care centers, in Ribeirão Preto, State of São Paulo, Brazil

A colonização da nasofaringe por Haemophilus influenzae (Hi) foi estudada em 114 crianças saudáveis com menos de 3 anos de idade e que freqüentam creches (day-care centers DCC) em Ribeirão Preto, estado de São Paulo, Brasil. Para cada uma das cepas isoladas foram determinados o biótipo, o sorotipo (por antisoro especifico e PCR) e a sensibilidade a 14 antibióticos. A freqüência de colonização por Hi foi de 72,0 por cento. As cepas isoladas foram identificadas como pertencentes aos biótipos II (36,5 po cento), I (21,5 por cento), V (18,2 por cento) e III (16,1 por cento). A freqüência encontrada de cepas encapsuladas foi de 3,2 por cento para o tipo f, 1,0 por cento para o tipo b, 1,0 por cento para o tipo d e 1,0 por cento para o tipo e. A resistência para trimetoprim-sulfametoxazole e ampicilina foi de 46,2 por cento e 10,7 por cento respectivamente. Resistência múltipla foi encontrada em 14 (15,0 por cento) das cepas analisadas. 13,9 por cento das cepas analisadas eram produtoras de b-lactamase, e não foi recuperada nenhuma cepa b-lactamase negativa e ampicilina resistente. DCCs são considerados locais de risco, com um alto potencial de disseminação de microrganismos e por isto devem ser continuadamente monitorados com a finalidade de detectar a eliminação da colonização da nasofaringe por cepas H. influenzae tipo b das crianças que freqüentam DCC, ou detectar a sua substituição por outro tipo de cepa.

Comparison of two slide agglutination serotyping methods and PCR-based capsule typing for the characterization of Haemophilus influenzae serotypes

Noventa e três isolados nasofaringeanos de H. influenzae foram sorotipados através de 2 métodos de aglutinação em lamina (SAST 1 e SAST 2) e os resultados foram comparados com a sorotipagem por PCR. SAST 1 apresentou uma baixa correlação com os resultados obtidos por PCR (75,2 por cento) enquanto que SAST 2 mostrou uma melhor concordância com os resultados da técnica molecular (93,5 por cento). Estes resultados indicam que SAST 2 pode ser um método alternativo para a correta detecção de H. influenzae tipo b.

Isolamento, caracterização e resistência a antimicrobianos de bactérias Gram-negativas aeróbias e anaeróbias facultativas de amostras de solo / Isolation, characterization, and antimicrobial resistance of Gram-negative aerobic and facultative anaerobic bacteria from soil samples

Cerca de 7% da população bacteriana do solo está representado por bactérias Gram-negativas aeróbias/anaeróbias facultativas. Muitas espécies são patogênicas e estão envolvidas em casos de infecções hospitalares. O potencial de patogenicidade dessas bactérias pode ser avaliado através da investigação da sua resistência a antimicrobianos. Além deste propósito, o objetivo do trabalho foi também identificar bacilos gram-negativos (BGN) isolados do solo. Para isso, 18 amostras de solo foram semeadas em caldo de Hajna. A obtenção de colônias isoladas seguiu-se em Mc Conkey ágar e Mueller Hinton ágar com 5% de sangue de carneiro. Foram repicadas 283 colônias com diferentes características morfológicas, em meio de IAL. Na triagem, obteve-se BGN fermentadores e não fermentadores, cuja identificação se fez pela metodologia convencional. Identificou-se 94,35% de enterobactérias e 5,65% BGN não fermentadores. A resistência bacteriana foi mais expressiva aos antibióticos ampicilina, cefalotina, cefoxitina, amoxicilina com ácido clavulânico e tetraciclina, variando entre 49,82% a 87,28%. Todos os isolados bacterianos foram resistentes a pelo menos um antibiótico, o que demonstra considerável potencial patogênico por constituírem um reservatório de resistência. A associação entre solo e resistência antimicrobiana precisa ser mais estudada. Observações e experimentos adicionais possibilitarão conhecer melhor essa relação e os processos que possam contribuir para a emergência de bactérias resistentes.

Bacterial diversity in soil in response to different plants, phosphate fertilizers and liming

The diversity of bacterial isolates from soil in response to different plants (control, Brachiaria ruziziensis and Cajanus cajan), fertilization (control, simple superphosphate and rock phosphate) and liming (with and without lime) was evaluated. Phenotypic and physiological characteristics of the isolates were recorded and organized in a file to identify the bacteria. Among the isolates, 95 percent were Gram-positive and 5 percent Gram-negative rods. Soil cultivated with B. ruziziensis favored the nonsporing Gram-positive and Gram-negative rods compared to soil with C. cajan or uncultivated. Number of spore-forming Gram-positive rods were higher in plots with superphosphate than in unfertilized soil or soil fertilized with rock phosphate. In unfertilized plots, larger number of Gram-positive cocci and Gram-negative rods was obtained than in fertilized plots. Unlimed plots favored spore-forming Gram-positive rods, Gram-positive cocci and Gram-negative rods, while liming a larger proportion of nonsporing Gram-positive rods was found. From 7 to 86 percent of the total isolates utilized different carbohydrates. The recording data used in this experiment was effective in the isolates identification, and might be useful for diagnosis of soil bacteria. Bacillus, Cellulomonas, Rhodococcus, Enterobacter, Flavobacterium, Micrococcus and Arthrobacter were the genera more commonly found. Bacterial diversity was enhanced in limed, unfertilized and plant cultivated plots.

Etiology of acute diarrhea among children in Ribeiräo Preto-SP, Brazil

To study the main enteropathogens causing diarrhea in the region of Ribeiräo Preto regarding serogroups and serotypes, the feces of 1836 children under 10 years old, from both sexes, attack of acute gastroenteritis, were analysed during a period of 4 years in Adolfo Lutz Institute - Ribeiräo Preto, SP. The pathogens identified by standard methods were the following: Escherichia coli, Salmonella spp., Shigella spp., Campylobacter spp., Yersinia spp., and Cryptosporidium spp. Positive samples were 22.8 percent (419) with 1.7 percent association of pathogens. Larger isolates were mainly from children 0 to 11 months old. Enteropathogenic E. coli (EPEC) was most frequent (8.7 percent) with predominance of serogroup O119 (40.2 percent), followed by Shigella (6.2 percent), 63.6 percent of which S. sonnei

Streptococcus pneumoniae and Haemophilus influenzae as etiological agents of conjunctivitis outbreaks in the region of Riveiräo Preto, SP, Brazil

No estudo de surtos de conjuntivite ocorridos no periodo de setembro de 1994 a setembro de 1996, na regiäo de Ribeirao Preto, foram semeadas no Instituto Adolfo Lutz Laboratorio I, Ribeiräo Preto, exsudatos conjuntivais de 92 pacientes, sendo que a maioria dos casos estava na faixa etaria de 2-7 anos. Os agentes etiologicos mais frequentemente isolados foram dos casos analisados foram: Streptococcus pneumoniae e Haemophilus influenzae em 40,22 por cento e 21,74 por cento respectivamente. 51,35 por cento das cepas de S. pneumoniae isoladas foram näo tipaveis. As cepas de S. pneumoniae oxacilina resistente foram submetidas ao teste de concentracao inibitoria minima (CIM), sendo que tres apresentaram resistencia intermediaria e apenas uma foi altamente resistente a penicilina