RESUMO
ABSTRACT Objective: Thymidine-dependent small-colony variants (TD-SCVs) of Staphylococcus aureus are being isolated with increasing frequency from patients with cystic fibrosis (CF). The aim of this study was to evaluate the relationship between TD-SCV isolation and pulmonary function in patients with CF, as well as to determine whether the emergence of TD-SCVs was associated with trimethoprim-sulfamethoxazole (TMP-SMX) use and with coinfection with other microorganisms. Methods: This was a retrospective case-control study including patients with CF who visited the Clinical Hospital Complex of the Federal University of Paraná, in Curitiba, Brazil, between 2013 and 2022. Demographic, clinical, and spirometric data, as well as information on TD-SCVs and other isolated microorganisms, were collected from the medical records of patients with CF and TD-SCVs (TD-SCV group; n = 32) and compared with those of a matched group of patients with CF without TD-SCVs (control group; n = 64). Results: Isolation of TD-SCVs was positively associated with TMP-SMX use (p = 0.009), hospitalization (p < 0.001), and impaired pulmonary function (p = 0.04). Conclusions: The use of TMP-SMX seems to contribute to the emergence of TD-SCVs, the isolation of which was directly associated with worse pulmonary function in our sample.
RESUMO
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.
Assuntos
Fibrose Cística/microbiologia , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/metabolismo , Timidina/metabolismo , Adolescente , Adulto , Idoso , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Brasil , Criança , Pré-Escolar , Farmacorresistência Bacteriana , Feminino , Humanos , Lactente , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Mutação , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/genética , Adulto JovemRESUMO
BACKGROUND: Small-colony variants (SCVs) are a morphologic subtype of Staphylococcus aureus that may occur through several mechanisms including auxotrophism for thymidine, hemin, or menadione. Auxotrophic SCV for thymidine fail to synthesize DNA specifically because of mutations in the thymidylate synthase gene. We isolated S. aureus thymidine-dependent SCVs (TD-SCV) from blood and respiratory samples of a pediatric patient with cystic fibrosis and pulmonary exacerbation. METHODS: Nutritional dependence of SCVs on hemin, menadione, and thymidine was evaluated. Antimicrobial susceptibility testing was performed through broth microdilution. Polymerase chain reaction was carried out for mecA, ermA, ermB, ermC, msrA, and msrB resistance genes. DNA sequencing was used to determine mutations in thyA and the multilocus sequence typing to identify genetic relatedness. RESULTS: Methicillin-sensitive S. aureus with normal and TD-SCV phenotypes were isolated from respiratory samples and a TD-SCV phenotype was isolated from blood culture. Macrolides resistance was attributed to ermC and msrB genes. All isolates belonged to ST398. The thyA gene in S. aureus is 957 nucleotides in length and encodes a protein of 318 amino acids. The TD-SCV isolates carried a -2 nt frameshift mutation (delta 667GC668) in thyA, creating a stop codon at residue 222 close to the predicted binding site for deoxyuridine monophosphate. CONCLUSIONS: The pathogenesis of SCVs is complex and not fully elucidated. Factors inherent to the patient such as physiological conditions, recurrent infections, or coinfection should be considered. Although SCVs are considered less virulent, they showed the ability to invade and cause bacteremia in the patient.
Assuntos
Bacteriemia/microbiologia , Fibrose Cística/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/genética , Staphylococcus aureus/isolamento & purificação , Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Genes Bacterianos , Variação Genética , Hemina , Humanos , Recém-Nascido , Masculino , Mutação , Fenótipo , Timidina , Vitamina K 3RESUMO
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.
Assuntos
Bacteriemia/microbiologia , Infecção Hospitalar/microbiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Unidades de Terapia Intensiva Neonatal , Ralstonia/isolamento & purificação , Bacteriemia/diagnóstico , Brasil , Infecção Hospitalar/diagnóstico , Feminino , Infecções por Bactérias Gram-Negativas/diagnóstico , Humanos , Recém-Nascido , MasculinoRESUMO
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.
Assuntos
Humanos , Masculino , Feminino , Recém-Nascido , Unidades de Terapia Intensiva Neonatal , Infecção Hospitalar/microbiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Bacteriemia/microbiologia , Ralstonia/isolamento & purificação , Brasil , Infecção Hospitalar/diagnóstico , Infecções por Bactérias Gram-Negativas/diagnóstico , Bacteriemia/diagnósticoRESUMO
The predisposition of patients with cystic fibrosis (CF) for recurrent pulmonary infections can result in poor prognosis of the disease. Although the clinical significance in CF of micro-organisms, such as Staphylococcus aureus, Haemophilus influenzae and Pseudomonas aeruginosa, is well established, the implication of uncommon glucose non-fermenting Gram-negative bacilli (UGNF-GNB) in respiratory samples from CF patients is still unclear. Because of limitations of traditional methods used in most clinical laboratories, the accurate identification of these microbes is a challenge. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) is an alternative tool for efficient identification of bacteria. This was a retrospective study to evaluate different identification methods in a collection of UGNF-GNB isolated from children with CF during a period of three years. The performance of MALDI-TOF was compared to that of 16S rDNA gene sequencing and to a conventional and automated phenotypic identification. The discriminatory power of MALDI-TOF (75.0â% agreement) was superior to automated techniques (67.1â% agreement) and to conventional phenotypical identification (50.0â% agreement). MALDI-TOF also demonstrated high accuracy in identifying Stenotrophomonas maltophilia, Achromobacter xylosoxidans and Chryseobacterium indologenes, but had limited utility in identifying Pandoraea spp. and some species of Acinetobacter and Chryseobacterium (other than C. indologenes). Although MALDI-TOF identified only 75â% of the isolates in comparison with 16S rDNA gene sequencing, the prompt identification and high discriminatory power exhibited by MALDI-TOF make it a useful tool for the characterization of micro-organisms that are difficult to identify using routine methods.
Assuntos
Técnicas Bacteriológicas/métodos , Fibrose Cística/complicações , Bactérias Gram-Negativas/química , Bactérias Gram-Negativas/classificação , Infecções por Bactérias Gram-Negativas/diagnóstico , Pneumonia Bacteriana/diagnóstico , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Criança , Pré-Escolar , Feminino , Bactérias Gram-Negativas/isolamento & purificação , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Lactente , Masculino , Pneumonia Bacteriana/microbiologia , Estudos RetrospectivosRESUMO
Há alguns anos tem-se verificado um aumento progressivo da resistência de alguns cocos gram-positivos a determinados antimicrobianos. Este aumento da resistência tem sido observado principalmente no ambiente hospitalar, e as bactérias mais comumente envolvidas são os Staphylococcus spp. e os Enterococcus spp. Devido a este fato, novos antimicrobianos são avaliados para o tratamento de infecções causadas por estas cepas multirresistentes. A associação quinupristina/dalfopristina (Q/D), também conhecida como Synercid©, é um antibacteriano da classe das estreptograminas, de uso endovenoso, composto por dois derivados semi-sintéticos da pristanamicina. A combinação das estreptograminas B e A na razão de 30:70 tem atividade antimicrobiana voltada para cocos gram-positivos, como Staphylococcus spp., Streptococcus spp., incluindo S. pneumoniae e Enterococcus faecium, sendo o E. faecalis habitualmente resistente. Neste estudo foi avaliada atividade in vitro de Q/D e outros oito antimicrobianos frente a 631 amostras de cocos gram-positivos isoladas de cinco centros brasileiros, complementadas com outras 20 cepas de E. faecium resistentes à vancomicina, provenientes dos Estados Unidos. Para a avaliação da sensibilidade aos antimicrobianos foi determinada a concentração inibitória mínima (MIC) pelo método do Etest (AB Biodisk, Solna, Suécia) e as cepas testadas foram: Staphylococcus aureus (n=267), Staphylococcus coagulase negativo (n=131), Streptoccus pneumoniae (n=130), Streptococcus beta-hemolíticos (n=28), Enteroccus faecalis (n=44) e E. faecium (n=51). A Q/D demonstrou excelente atividade contra Staphylococcus spp., independente de serem sensíveis ou resistentes à oxacilina. Para S. pneumoniae, a Q/D apresentou igualmente uma ótima atividade, inclusive para as cepas com resistência intermediária ou total para penicilina. Entre as cepas de E. faecium sensíveis à vancomicina, o MIC 90 de Q/D obtido foi de 3µg/ml, sendo que 45 por cento das cepas testadas foram sensíveis e 55 por cento apresentaram sensibilidade intermediária à associação. Desta forma, pode-se afirmar que a associação. Desta forma, pode-se afirmar que a associação Q/D representa uma nova opção para o tratamento endovenoso de infecções causadas por cocos gram-positivos, principalmente para as cepas multiresistentes, sendo também uma alternativa ao uso de glicopeptídeos
Assuntos
Antibacterianos , Cocos Anaeróbios Gram-Negativos , Quimioterapia Combinada , Especificidade da Espécie , Técnicas In Vitro , Testes de Sensibilidade Microbiana , Resistência a Múltiplos Medicamentos , Resistência Microbiana a Medicamentos , América LatinaRESUMO
We report a fatal case of septicemia due to a vancomycin-resistant Enterococcus faecium in a 9 year-old girl with aplastic anemia. The isolate was also resistant to amplicillin, teicoplanin, gentamicin (high level), and streptomycin (high level). We believe that this is the first case of vancomycin-resistant Enterococcus (VRE) reported from a clinical specimen in Brazil.