Comparative analysis of the antimicrobial resistance and virulence traits in ESBL-producing-Klebsiella pneumoniae ST307 strains colonizing the gastrointestinal tract and causing a fatal bloodstream infection in a leukemia patient.

Klebsiella pneumoniae is an opportunistic pathogen that can colonize the gastrointestinal tract (GIT) of humans. The mechanisms underlying the successful translocation of this pathogen to cause extra-intestinal infections remain unknown, although virulence and antimicrobial resistance traits likely play significant roles in the establishment of infections. We investigated K. pneumoniae strains isolated from GIT colonization (strains Kp_FZcol-1, Kp_FZcol-2 and Kp_FZcro-1) and from a fatal bloodstream infection (strain Kp_HM-1) in a leukemia patient. All strains belonged to ST307, carried a transferable IncF plasmid containing the blaCTX-M-15 gene (pKPN3-307 TypeA-like plasmid) and showed a multidrug-resistance phenotype. Phylogenetic analysis demonstrated that Kp_HM-1 was more closely related to Kp_FZcro-1 than to the other colonizing strains. The Kp_FZcol-2 genome showed 81 % coverage with the Kp_HM-1 246,730 bp plasmid (pKp_HM-1), lacking most of its putative virulence genes. Searching public genomes with similar coverage, we observed the occurrence of this deletion in K. pneumoniae ST307 strains recovered from human colonization and infection in different countries. Our findings suggest that strains lacking the putative virulence genes found in the pKPN3-307 TypeA plasmid are still able to colonize and infect humans, highlighting the need to further investigate the role of these genes for the adaptation of K. pneumoniae ST307 in distinct human body sites.

Colistin resistance emerges in pandrug-resistant Klebsiella pneumoniae epidemic clones in Rio de Janeiro, Brazil.

Klebsiella pneumoniae is an important human pathogen, able to accumulate and disseminate a variety of antimicrobial resistance genes. Resistance to colistin, one of the last therapeutic options for multi-drug-resistant bacteria, has been reported increasingly. Colistin-resistant K. pneumoniae (ColRKp) emerged in two hospitals in Rio de Janeiro state, Brazil in 2016. The aim of this study was to investigate if these ColRKp isolates were clonally related when compared between hospitals, to identify the molecular mechanisms of colistin resistance, and to describe other antimicrobial resistance genes carried by isolates. Twenty-three isolates were successively recovered, and the whole-genome sequence was analysed for 10, each of a different pulsed-field gel electrophoresis (PFGE) type. Although some PFGE clusters were found, none of them included isolates from both hospitals. Half of the isolates were assigned to CC258, three to ST152 and two to ST15. One isolate was pandrug resistant, one was extensively drug resistant, and the others were multi-drug resistant. Colistin resistance was related to mutations in mgrB, pmrB, phoQ and crrB. Eleven new mutations were found in these genes, including two nucleotide deletions in mgrB. All isolates were carbapenem resistant, and seven were associated with carbapenemase carriage (blaKPC-2 in six isolates and blaOXA-370 in one isolate). All isolates had a blaCTX-M, and two had a 16S ribosomal RNA methyltransferase encoding gene (armA and rmtB). ColRKp were composed of epidemic clones, but cross-dissemination between hospitals was not detected. Colistin resistance emerged with several novel mutations amid highly resistant strains, further restricting the number of drugs available and leading to pandrug resistance.

Major globally disseminated clonal complexes of antimicrobial resistant enterococci associated with infections in cancer patients in Brazil.

Cancer and hematological malignancies constitute major comorbidities in enterococcal infections, but little is known about the characteristics of enterococci affecting cancer patients. The aim of this study was to characterize 132 enterococcal clinical isolates obtained from cancer patients attending a Cancer Reference Center in Brazil between April 2013 and March 2014. Susceptibility to 17 antimicrobial agents was assessed by disk diffusion method. Resistance and virulence genes were investigated by PCR. Multilocus sequence typing (MLST) was performed for selected Enterococcus faecalis and Enterococcus faecium isolates. The predominant species was E. faecalis (108 isolates), followed by E. faecium (18), Enterococcus gallinarum (3), Enterococcus avium (2) and Enterococcus durans (1). Multidrug-resistant (MDR) isolates made up 44.7%, but all isolates were susceptible to fosfomycin, linezolid and glycopeptides. The most prevalent genes associated with erythromycin- and tetracycline-non susceptible isolates were erm(B) (47/71; 66.2%) and tet(M) (24/68; 35.3%), respectively. High-level resistance (HLR) to gentamicin was found in 22 (16.7%) isolates and 13 (59.1%) of them carried the aac(6')-Ie-aph(2″)-Ia gene. HLR to streptomycin was detected in 34 (25.8%) isolates, of which 15 (44.1%) isolates had the ant(6')-Ia gene. The most common virulence genes were gelE (48.9%), esp (30.5%) and asa1 (29.8%). MLST performed for 26 E. faecalis isolates revealed 18 different sequence-types (STs), with seven corresponding to novel STs (625, 626, 627, 628, 629, 630, and 635). On the other hand, nine of 10 E. faecium isolates analyzed by MLST belonged to a single clonal complex, comprised of mostly ST412, which emerged worldwide after mid-2000s, but also two novel STs (963 and 964). We detected major globally disseminated E. faecalis and E. faecium clonal complexes along with novel closely related STs, indicating the fitness and continuous evolution of these hospital-adapted lineages.

Serotypes, antimicrobial resistance and genotypes of Streptococcus pneumoniae associated with infections in cancer patients in Brazil.

We sought to characterize pneumococcal isolates associated with bacteremia, pneumonia and meningitis in cancer patients and to estimate the coverage of the available pneumococcal vaccines. Fifty isolates recovered from 49 patients attending a cancer reference center over a 1-year period were analyzed. The prevalent serotypes were: 23F (12%), 6A (8%), 3, 4, 20, and 23A (6% each). All isolates were susceptible to chloramphenicol, levofloxacin, rifampicin, and vancomycin. Resistance or reduced susceptibility to penicillin made up 14%, and one isolate was also intermediately resistant to ceftriaxone. The three (6%) erythromycin-resistant isolates presented the M or cMLSB phenotypes and harbored the mef(A/E) gene exclusively or along with the erm(B) gene. Twenty-two (44%) isolates were closely related to 11 international clones, being strongly associated with penicillin non-susceptibility. Combined immunization with the 13-valent conjugate and the 23-valent polysaccharide vaccines might contribute to reduce (76%) the burden of the pneumococcal infections in the population investigated.

Risk factors for and mortality of extended-spectrum-beta-lactamase-producing Klebsiella pneumoniae and Escherichia coli nosocomial bloodstream infections.

A case-control study, involving patients with positive blood cultures for Klebsiella pneumoniae (KP) or Escherichia coli (EC) EC and controls with positive blood cultures for non-ESBL-KP or EC, was performed to assess risk factors for extended-spectrum-beta-lactamase (ESBL) production from nosocomial bloodstream infections (BSIs). Mortality among patients with BSIs was also assessed. The study included 145 patients (81, 59.5% with K. pneumoniae and 64, 44.1% with E. coli BSI); 51 (35.2%) isolates were ESBL producers and 94 (64.8%) nonproducers. Forty-five (55.6%) K. pneumoniae isolates were ESBL producers, while only six (9.4%) E. coli isolates produced the enzyme. Multivariate analysis showed that recent exposure to piperacillin-tazobactam (adjusted Odds Ratio [aOR] 6.2; 95%CI 1.1-34.7) was a risk factor for ESBL BSI. K. pneumoniae was significantly more likely to be an ESBL-producing isolate than E. coli (aOR 6.7; 95%CI 2.3-20.2). No cephalosporin class was independently associated with ESBLs BSI; however, in a secondary model considering all oxymino-cephalosporins as a single variable, a significant association was demonstrated (aOR 3.7; 95%CI 1.3-10.8). Overall 60-day mortality was significantly higher among ESBL-producing organisms. The finding that piperacillin-tazobactam use is a risk factor for ESBL-production in KP or EC BSIs requires attention, since this drug can be recommended to limit the use of third-generation cephalosporins.

Clinical and molecular epidemiology of methicillin-resistant Staphylococcus aureus carrying SCCmecIV in a university hospital in Porto Alegre, Brazil.

We evaluated clinical outcomes and molecular epidemiology of methicillin-resistant Staphylococcus aureus carrying SCCmecIV recovered from patients who attended at a teaching hospital from Porto Alegre, Brazil. All Panton-Valentine leukocidin (PVL)-producer isolates belonged to clonal complex (CC) 30 (11 isolates, related to Oceania Southwest Pacific clone [OSPC]), and the PVL-negative isolates were typed as CC5 (2 isolates, related to the pediatric clone). Five patients had health care-associated infections (HCAIs) with hospital-onset, 5 HCAIs with community-onset, and 3 community-acquired infections without risks. A high overall mortality (30.8%) was found. This study show that OSPC isolates are not only causing community-associated infections but are also involved in HCAI in our country.

Risk factors for and mortality of extended-spectrum-beta-lactamase-producing Klebsiella pneumoniae and Escherichia coli nosocomial bloodstream infections / Fatores de risco e mortalidade de infecções da corrente sanguínea por Klebsiella pneumoniae and Escherichia coli produtores de beta-lactamase de espectro estendido

A case-control study, involving patients with positive blood cultures for Klebsiella pneumoniae (KP) or Escherichia coli (EC) EC and controls with positive blood cultures for non-ESBL-KP or EC, was performed to assess risk factors for extended-spectrum-β-lactamase (ESBL) production from nosocomial bloodstream infections (BSIs). Mortality among patients with BSIs was also assessed. The study included 145 patients (81, 59.5 percent with K. pneumoniae and 64, 44.1 percent with E. coli BSI); 51 (35.2 percent) isolates were ESBL producers and 94 (64.8 percent) nonproducers. Forty-five (55.6 percent) K. pneumoniae isolates were ESBL producers, while only six (9.4 percent) E. coli isolates produced the enzyme. Multivariate analysis showed that recent exposure to piperacillin-tazobactam (adjusted Odds Ratio [aOR] 6.2; 95 percentCI 1.1-34.7) was a risk factor for ESBL BSI. K. pneumoniae was significantly more likely to be an ESBL-producing isolate than E. coli (aOR 6.7; 95 percentCI 2.3-20.2). No cephalosporin class was independently associated with ESBLs BSI; however, in a secondary model considering all oxymino-cephalosporins as a single variable, a significant association was demonstrated (aOR 3.7; 95 percentCI 1.3-10.8). Overall 60-day mortality was significantly higher among ESBL-producing organisms. The finding that piperacillin-tazobactam use is a risk factor for ESBL-production in KP or EC BSIs requires attention, since this drug can be recommended to limit the use of third-generation cephalosporins.

Estudo de caso-controle, onde os casos foram pacientes com hemocultura positiva para Klebsiella pneumoniae (KP) ou Escherichia coli (EC) produtores de beta lactamase de espectro estendido (ESBL) e os controles foram pacientes com hemoculturas positivas para EC ou KP não produtores de ESBL foi realizado para avaliar os fatores de risco para produção destas enzimas em infecções da corrente sanguínea (ICS). Mortalidade dos pacientes com ICS também foi avaliada. Foram incluídos 145 pacientes (81, 59,5 por cento tinham Klebsiella pneumoniae e 64, 44,1 por cento tinham Escherichia coli); 51 (35,2 por cento) isolados eram produtores de ESBL e 94 (64,8 por cento) eram não produtores. Quarenta e cinco (55,6 por cento) isolados de K. pneumoniae e somente seis (9,4 por cento) isolados de E. coli eram produtores de ESBL. Análise multivariada mostrou que exposição recente a piperacilina-tazobactam (OR ajustado [aOddsRatio] 6,2; 95 por cento Intervalo de Confiança [IC] 1,1-34,7) foi fator de risco para infecção da corrente sanguínea por ESBL. Foi significativamente maior a chance de K. pneumoniae ser um isolado produtor de ESBL do que E. coli o ser (aOR 6,7; 95 por cento CI 2,3-20,2). Nenhuma classe de cefalosporina foi independentemente associada com ESBL-ICS. No entanto, em um modelo secundário considerando todas as oximino-cefalosporinas como variável única, foi demonstrada associação significativa (aOR 3,7; 95 por cento IC 1,3-10,8). Mortalidade total em 60 dias foi significativamente maior entre isolados produtores de ESBL. O achado de piperacilina-tazobactam como fator de risco para produção de ESBL em ICS por KP ou EC requer atenção, uma vez que esta droga tem sido eventualmente recomendada para poupar o uso de cefalosporinas de terceira geração.

Indications of carbapenem resistance evolution through heteroresistance as an intermediate stage in Acinetobacter baumannii after carbapenem administration.

We describe an in vivo evolution of an antimicrobial profile from susceptibility to full-resistance to carbapenems, with heteroresistance as an intermediate stage, in an Acinetobacter baumannii strain. Heteroresistance was characterized by the growth of sub-populations within the susceptibility halo in both disk-diffusion and Etest. PCRs for the main A. baumannii carbapenemases were negative. The exact resistance mechanism, diagnostic methods and clinical relevance of heteroresistance in A. baumannii warrant further investigations. This is the first description of such phenomenon in vivo and the second report of heteroresistance to carbapenems in A. baumannii.

Corynebacterium striatum infecting a malignant cutaneous lesion: the emergence of an opportunistic pathogen.

We described a case of a 27-year old male patient with skin and soft tissue infection of a neoplastic lesion caused by Corynebacterium striatum, an organism which has been rarely described as a human pathogen. Identification was confirmed by DNA sequencing. Successful treatment with penicillin was achieved. The role of the C. striatum as an emerging opportunistic pathogen is discussed.

In vitro fosfomycin activity in vancomycin-resistant Enterococcus faecalis

Enterococci are part of the endogenous flora of human beings, are naturally resistant to several classes of antimicrobials, and are able to acquire resistance with relative ease. Currently the vancomycin-resistant enterococci are spread all over the world and treatment options for infections caused by it are often extremely limited. We assessed 193 vancomycin-resistant Enterococcus faecalis isolates collected from four different hospitals in Porto Alegre for their susceptibility to fosfomycin using the E-test and agar diffusion. Fosfomycin proved to be active in vitro against the great majority of isolates, indicating that it is a valid option in the treatment of these infections.

Indications of carbapenem resistance evolution through heteroresistance as an intermediate stage in Acinetobacter baumannii after carbapenem administration / Indicações da evolução da resistência ao carbapenem através da heteroresistência como estágio intermediário no Acinetobacter baumannii após administração de carbapenem

We describe an in vivo evolution of an antimicrobial profile from susceptibility to full-resistance to carbapenems, with heteroresistance as an intermediate stage, in an Acinetobacter baumannii strain. Heteroresistance was characterized by the growth of sub-populations within the susceptibility halo in both disk-diffusion and Etest. PCRs for the main A. baumannii carbapenemases were negative. The exact resistance mechanism, diagnostic methods and clinical relevance of heteroresistance in A. baumannii warrant further investigations. This is the first description of such phenomenon in vivo and the second report of heteroresistance to carbapenems in A. baumannii.

Descrevemos a evolução in vivo, de um perfil de sensibilidade aos antimicrobianos, passando de sensibilidade a resistência total aos antibióticos carbapenêmicos, com um estágio intermediário de heteroresistência em isolado de Acinetobacter baumannii. A heteroresistência foi caracterizada pelo crescimento de sub-população na zona de inibição pelo método de disco-difusão e pelo Etest. PCRs para as principais carbapenemases envolvidas com resistência neste microrganismo foram negativas. O exato mecanismo de resistência envolvido, método diagnóstico e relevância clínica justificam investigação adicional. Esta é a primeira descrição deste fenômeno in vivo e o segundo relato de heteroresistência em A. baumannii.

Corynebacterium striatum infecting a malignant cutaneous lesion: the emergence of an opportunistic pathogen / Corynebacterium striatum infectando lesão cutânea maligna: a emergência de um patógeno oportunista

We described a case of a 27-year old male patient with skin and soft tissue infection of a neoplastic lesion caused by Corynebacterium striatum, an organism which has been rarely described as a human pathogen. Identification was confirmed by DNA sequencing. Successful treatment with penicillin was achieved. The role of the C. striatum as an emerging opportunistic pathogen is discussed.

Descrevemos infecção de lesão neoplásica em paciente masculino de 27 anos, envolvendo pele e partes moles, causada por Corynebacterium striatum, um microrganismo raramente descrito como patógeno humano. A identificação foi confirmada por seqüenciamento de DNA. O paciente foi tratado com penicilina, com sucesso. O papel do C. striatum como patógeno oportunista é discutido.

In vitro fosfomycin activity in vancomycin-resistant Enterococcus faecalis.

Enterococci are part of the endogenous flora of human beings, are naturally resistant to several classes of antimicrobials, and are able to acquire resistance with relative ease. Currently the vancomycin-resistant enterococci are spread all over the world and treatment options for infections caused by it are often extremely limited. We assessed 193 vancomycin-resistant Enterococcus faecalis isolates collected from four different hospitals in Porto Alegre for their susceptibility to fosfomycin using the E-test and agar diffusion. Fosfomycin proved to be active in vitro against the great majority of isolates, indicating that it is a valid option in the treatment of these infections.

Pharmacokinetics of intravenous polymyxin B in critically ill patients.

BACKGROUND: Although not much pharmacokinetic knowledge is available, polymyxin B is increasingly used for treatment of infections caused by gram-negative bacteria that are resistant to all other antibiotics. METHODS: This study involved 8 patients who received intensive care after intravenous administration of a 60-min infusion of polymyxin B at currently recommended doses. Blood and urine samples were collected, and plasma protein binding of polymyxin B was determined. Concentrations of polymyxin B in plasma and urine samples were measured by a specific high-performance liquid chromatographic method. RESULTS: Polymyxin B was well tolerated. The peak plasma concentrations at the end of the infusion varied from 2.38 to 13.9 mg/L. For 4 patients from whom it was possible to collect urine samples over a dosing interval, only 0.04%-0.86% of the dose was recovered in the urine in unchanged form. Plasma protein binding of polymyxin B was higher in samples from patients (range, 78.5%-92.4%) than in plasma samples from healthy human subjects (mean +/- standard deviation, 55.9% +/- 4.7%). Unbound plasma concentrations of polymyxin B were in the vicinity of or lower than the minimum inhibitory concentration of the pathogen. CONCLUSION: To our knowledge, this is the first study to report plasma concentrations over time and urinary recovery of polymyxin B in critically ill patients after intravenous administration. Polymyxin B is eliminated mainly by nonrenal pathways, and the total body clearance appears to be relatively insensitive to renal function. Additional investigations are required to assess the appropriateness of currently recommended doses of this drug for the treatment of severe infections in critically ill persons.

Hiper-infecção por Strongyloides Stercoralis: relato de caso / Strongyloides Stercoralis hyperinfection syndrome: case report

JUSTIFICATIVA E OBJETIVOS: O Strongyloides stercoralis é um agente comum de infecção do trato gastrintestinal. Em pacientes imunodeprimidos este nematódeo pode causar hiper-infecção, com manifestações pulmonares e sepse por germes gram-negativos. O objetivo deste relato foi apresentar um caso com evolução letal e ressaltar a importância do diagnóstico e do tratamento precoce. RELATO DO CASO: Paciente do sexo masculino, 60 anos, com diagnóstico de tumor de timo, submetido a tratamento cirúrgico, radioterapia e quimioterapia. Foi consultado na emergência relatando queixa de diarréia e dispnéia, sendo admitido na UTI após apresentar quadro de insuficiência respiratória aguda hipoxêmica e choque refratário, evoluindo para óbito. No aspirado traqueal, foi identificado larvas de Strongyloides stercoralis. CONCLUSÕES: A estrongiloidíase, apesar de tratar-se de infecção parasitária freqüentemente leve, em pacientes imunodeprimidos pode apresentar-se de forma grave e disseminada. Deve-se suspeitar deste agente em pacientes que vivem em áreas endêmicas, sendo o diagnóstico estabelecido através da pesquisa da larva do Strongyloides stercoralis na secreção traqueal e nas fezes.

BACKGROUND AND OBJECTIVES: Strongyloides Stercoralis is a common cause of gastrointestinal infection. This nematode can produce an overwhelming hyperinfection syndrome, especially in the immunocompromised patient. Typically, patients present with pulmonary symptoms, but subsequently they can acquire Gram-negative sepsis. The objective of this report is to describe a lethal case and call attention to the importance of early diagnosis and treatment. CASE REPORT: Male patient, 60 year-old with diagnosis of timoma, treated with surgery, radiotherapy and chemotherapy in the past. He presented to the emergency room complaining of diarrhea and dyspnea, and then transferred to the ICU after development of hypoxemic acute respiratory failure and refractory septic shock, and despite treatment the patient died. A bronchial sample of sputum showed Strongyloides stercoralis worms. CONCLUSIONS: Strongyloides stercoralis infection symptoms are usually mild, but in the setting of impaired host immunity, a disseminated and severe illness may occur. Clinicians must be aware for patients from endemic areas. Diagnosis may be established through sputum and stool examination for Strongyloides stercoralis worms.

[Strongyloides Stercoralis hyperinfection syndrome: case report]. / Hiper-infecção por Strongyloides Stercoralis: relato de caso.

BACKGROUND AND OBJECTIVES: Strongyloides Stercoralis is a common cause of gastrointestinal infection. This nematode can produce an overwhelming hyperinfection syndrome, especially in the immunocompromised patient. Typically, patients present with pulmonary symptoms, but subsequently they can acquire Gram-negative sepsis. The objective of this report is to describe a lethal case and call attention to the importance of early diagnosis and treatment. CASE REPORT: Male patient, 60 year-old with diagnosis of timoma, treated with surgery, radiotherapy and chemotherapy in the past. He presented to the emergency room complaining of diarrhea and dyspnea, and then transferred to the ICU after development of hypoxemic acute respiratory failure and refractory septic shock, and despite treatment the patient died. A bronchial sample of sputum showed Strongyloides stercoralis worms. CONCLUSIONS: Strongyloides stercoralis infection symptoms are usually mild, but in the setting of impaired host immunity, a disseminated and severe illness may occur. Clinicians must be aware for patients from endemic areas. Diagnosis may be established through sputum and stool examination for Strongyloides stercoralis worms.