Candida haemulonii is a complex formed by C. haemulonii sensu stricto , C. haemulonii var. vulnera , and C. duobushaemulonii . Members of this complex are opportunistic pathogens closely related to C. pseudohaemulonii , C. lusitaniae , and C. auris , all members of a multidrug-resistant clade. Complete genome sequences for all members of this group are available in the GenBank database, except for C. haemulonii var. vulnera . Here, we report the first draft genomes of two C. haemulonii var. vulnera (isolates K1 and K2) and comparative genome analysis of closely related fungal species. The isolates were biofilm producers and non-susceptible to amphotericin B and fluconazole. The draft genomes comprised 350 and 387 contigs and total genome sizes of 13.21 and 13.26 Mb, with 5,479 and 5,507 protein-coding genes, respectively, allowing the identification of virulence and resistance genes. Comparative analyses of orthologous genes within the multidrug-resistant clade showed a total of 4,015 core clusters, supporting the conservation of 24,654 proteins and 3,849 single-copy gene clusters. Candida haemulonii var. vulnera shared a larger number of clusters with C. haemulonii and C. auris ; however, more singletons were identified in C. lusitaniae and C. auris . Additionally, a multiple sequence alignment of Erg11p proteins revealed variants likely involved in reduced susceptibility to azole and polyene antifungal agents. The data presented in this work will, therefore, be of utmost importance for researchers studying the biology of the C. haemulonii complex and related species.

Aim The current study was conducted to determine the antimicrobial resistance profile and genetic relatedness of Aeromonas sp. isolated from healthcare and urban effluents, wastewater treatment plant (WWTP) and river water. Methods and Results We detected the presence of genes conferring resistance to β‐lactam, quinolone and aminoglycoside. Multilocus sequence typing was carried out to differentiate the strains, and multilocus phylogenetic analysis was used to identify the species. A total of 28 cefotaxime‐resistant Aeromonas sp. strains were identified, harbouring uncommon Guiana‐extended‐spectrum (GES)‐type β‐lactamases (GES‐1, GES‐5, GES‐7 and GES‐16). Multidrug‐resistant Aeromonas sp. were found in hospital wastewater, WWTP and sanitary effluent, and A. caviae was identified as the most prevalent species (85·7%). Conclusion The release of untreated healthcare effluents, presence of antimicrobials in the environment, in addition to multidrug‐resistant Aeromonas sp., are all potential factors for the spread of resistance. Significance and Impact of the Study We identified a vast repertoire of antimicrobial resistance genes (ARG) in Aeromonas sp. from diverse aquatic ecosystems, including those that encode enzymes degrading broad‐spectrum antimicrobials widely used to treat healthcare‐associated infections. Hospital and sanitary effluents serve as potential sources of bacteria harbouring ARG and are a threat to public health.

Background: Invasive candidiasis (IC) is a major cause of morbimortality in children. Previous studies described the clinical characteristics and risk factors for this infection; however, limited data are available on the predictors of mortality in these patients. In this context, we evaluated the risk factors associated with death due to IC in a pediatric tertiary care hospital in South of Brazil. Methods: This is a retrospective, cross-sectional, observational, and analytical study of a series of pediatric patients with clinical and laboratory diagnosis of IC from March 2014 to September 2017. Univariate and multivariate analysis were performed to estimate the association between the characteristics of the patients and death. Results: A total of 94 cases of IC were included. The incidence was 1.13 cases per 1000 patients/d, with a mortality rate of 14%. There was a predominance of non-albicans Candida (71.3%) in IC cases and, although there is no species difference in mortality rates, biofilm formation was associated with increased mortality. Clinical characteristics such as male sex, stay in the intensive care unit, and thrombocytopenia; comorbidities such as cardiological disease and renal insufficiency; and risks such as mechanical ventilation and dialysis were associated with increased mortality. Conclusion: Data from this study suggest that biofilm formation by Candida sp. is associated with increased mortality, and this is the first study to correlate the male sex and cardiological disease as risk factors for death in pediatric IC patients.

Introdução: Stenotrophomonas maltophilia é um importante patógeno hospitalar emergente, naturalmente resistente aos carbapenêmicos, que possui sulfametoxazol/trimetoprima (SMX-TMP) como opção terapêutica. Porém, relatos de resistência a essa droga começaram a aparecer. Objetivos: Avaliar o perfil de sensibilidade das cepas de S. maltophilia e comparar os resultados obtidos por diferentes metodologias. Método: As cepas isoladas na rotina microbiológica do Hospital das Clínicas de São Paulo em 2007 foram avaliadas quanto ao perfil de suscetibilidade por disco-difusão e microdiluição. Respeitando a padronização do Clinical and Laboratory Standard Institute (CLSI), SMX-TMP e levofloxacino foram testados pelas duas técnicas, bem como a tigeciclina, interpretada segundo a agência norte-americana Food and Drug Administration (FDA). Ticarcilina/clavulanato, ceftazidima, imipenem, meropenem e moxifloxacino foram avaliados apenas por microdiluição. O percentual de correlação entre as metodologias foi analisado pelo programa WHONET ® . Resultados: Das 126 cepas, 1,6% apresentaram resistência a SMX-TMP; 2,4%, a levofloxacino; 23%, a ticarcilina/clavulanato; 54%, a ceftazidima. Todas apresentaram concentração inibitória mínima (CIM) < 2 µg/mL para tigeciclina e 96,8%, CIM ≤ 2 µg/mL para moxifloxacino. O índice de concordância categórica entre as metodologias foi de 100% para SMX-TMP e de 89,6% para levofloxacino. Discussão e conclusão: Embora SMX-TMP seja a droga de escolha para o tratamento de infecções por S. maltophilia, há possibilidade de resistência a esse antibiótico, dificultando a decisão terapêutica empírica, sendo fundamental a realização do teste de sensibilidade. A técnica de disco-difusão demonstrou boa correlação com a microdiluição. Entre as novas opções terapêuticas, tigeciclina e moxifloxacino apresentaram boa atividade in vitro. resumo unitermos Stenotrophomonas maltophiliaDisco-difusão Microdiluição Sulfametoxazol/ trimetoprima abstract Introduction: Stenotrophomonas maltophilia is an important emerging nosocomial pathogen naturally resistant to carbapenems. Although trimethoprim/sulfamethoxazole (TMP-SMX) is commonly used as a treatment option, resistance to this drug has been recently reported. Objectives: To evaluate the susceptibility profile of S. maltophilia strains and compare the results obtained by different methods. Methods: As to susceptibility profile, all strains, which had been routinely isolated at Hospital das Clínicas, São Paulo, in 2007, were evaluated through disk diffusion and microdilution. In accordance with the Clinical and Laboratory Standard Institute (CLSI), TMP-SMX and levofloxacin were tested through both techniques, as well as tigecycline, which was interpreted in conformity with Food and Drug Administration (FDA) regulations. Ticarcillin/clavulanate, ceftazidime, imipenem, meropenem and moxifloxacin were evaluated only through microdilution. The correlation between these methods was assessed by WHONET ® . Results: 1.6% of 126 strains were resistant to TMP-SMX, 2.4% to levoflo...

Introduction. This study aimed to evaluate different methods for differentiation of species of coagulase-negative staphylococci (CoNS) that caused infections in hospitalized immunocompromised patients. Methods. A total of 134 CoNS strains were characterized using four different methods. Results. The results of matrix assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) analysis were in complete agreement with those of tuf gene sequencing (kappa index = 1.00 . The increasing incidence of CoNS hospitalacquired infections emphasizes the need for accurate and rapid identification of Staphylococcus at the species level. Although the ability to distinguish between these pathogens is clinically relevant, a number of laboratory limitations still exist 1,3 . Molecular methods are currently favored for accurate identification and several target genes, such as tuf gene, exhibit high discriminatory power 4 . The aim of this study was to compare the ability of matrix-assisted laser desorption/ionization timeof-flight mass spectrometry (MALDI-TOF MS) (Microflex LT instrument, Bruker Daltonics, Bremen, Germany) and three other identification methods to differentiate between CoNS samples.A total of 134 CoNS clinical isolates from the blood cultures (112) and catheter tips (22) of patients hospitalized in a tertiary care teaching Hospital in Curitiba, Brazil were studied. The discriminatory power of MALDI-TOF MS was compared to that of the gold standard tuf gene sequencing method, an automated method (Vitek ® 2 Compact, bioMérieux, Marcy l'Etoile, France), and a conventional method for all CoNS included in the study. In addition, the minimum inhibitory concentrations (MICs) of the antimicrobials oxacillin, daptomycin, linezolid, and vancomycin were evaluated and the presence of the mecA gene was determined by PCR for all strains.All bacterial isolates were preliminarily identified by catalase, coagulase, and deoxyribonuclease tests. In addition, the disk diffusion assay was used to determine susceptibility to fosfomycin (200µg) and desferrioxamine (300µg) 5 . Subsequently, the strains were evaluated using the conventional method proposed by Cunha et al., for distinguishing between species not identified by the previous tests 3 . S. epidermidis ATCC 12228 was used as the control strain. Finally, the Vitek ® 2 Compact automated system was used as per the manufacturer's instructions; the acceptable criterion for identification was set at 93-99% probability.For MALDI-TOF MS analysis, a loopful of cells from a fresh overnight pure culture was processed as previously described 6 . The obtained mass spectra were analyzed using the Microflex LT instrument with MALDI Biotyper 3.0 software. The identification score cutoff value was set at ≥1.8 7 .