LAURO VIEIRA PERDIGAO NETO

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LIM/49 - Laboratório de Protozoologia, Hospital das Clínicas, Faculdade de Medicina

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  • article 21 Citação(ões) na Scopus
    Multidrug-resistant Stenotrophomonas maltophilia: Description of new MLST profiles and resistance and virulence genes using whole-genome sequencing
    (2018) RIZEK, Camila Fonseca; JONAS, Daniel; PAEZ, Jorge Isaac Garcia; ROSA, Juliana Ferraz; PERDIGAO NETO, Lauro Vieira; MARTINS, Roberta Ruedas; MORENO, Luisa Z.; ROSSI JUNIOR, Alfio; LEVIN, Anna S.; COSTA, Silvia Figueiredo
    Objectives: Stenotrophomonas maltophilia is an opportunistic pathogen that has high intrinsic and acquired antimicrobial resistance, with great genetic diversity. The aim of this study was to characterise four S. maltophilia clinical isolates displaying different susceptibility profiles using whole-genome sequencing. Methods: The whole genomes of four clinical isolates of S. maltophilia from three patients were sequenced using Ion Torrent (TM) PGM technology. The isolates presented different susceptibilities to trimethoprim/sulfamethoxazole (SXT) and levofloxacin. Results: Three new multilocus sequence typing (MLST) profiles were identified (ST144, ST172 and ST173), differing in virulence and resistance genes. The ST172 isolate had more genes related to toxins than related to motility or adhesion and had different types of efflux pumps than the other isolates. The SXT-resistant strains belonged to ST172 or ST144 and did not harbour the sul1, sul2 or dfrA resistance genes. Strains I and II, from the same patient and belonging to the same ST but differing in resistance to SXT, had all of the resistance genes searched for in common, except for the SmeABC efflux pump complex genes that were only found in the SXT-resistant strain. All strains, including the strain susceptible to levofloxacin, harboured the qnrB gene, which may question the importance of this gene in determining levofloxacin resistance in S. maltophilia. Conclusion: Here we describe three new MLST profiles. Resistance to SXT in these strains appears to be associated with efflux pumps.
  • conferenceObject
    Current Status of Antimicrobial Stewardship Programs in Sao Paulo Hospitals
    (2020) SATO, Silvia; MADALOSSO, Geraldine; ASSIS, Denise; PERDIGAO-NETO, Lauro; LEVIN, Anna; OLIVEIRA, Maura
  • article 12 Citação(ões) na Scopus
    Alternative drugs against multiresistant Gram-negative bacteria
    (2020) PERDIGAO NETO, Lauro Vieira; OLIVEIRA, Maura Salaroli; ORSI, Tatiana D'Annibale; PRADO, Gladys Villas Boas do; MARTINS, Roberta Cristina Ruedas; LEITE, Gleice Cristina; MARCHI, Ana Paula; LIRA, Esther Sant'Ana de; CORTES, Marina Farrel; ESPINOZA, Evelyn Patricia Sanchez; CARRILHO, Claudia Maria Dantas de Maio; BOSZCZOWSKI, Icaro; GUIMARAES, Thais; COSTA, Silvia Figueiredo; LEVIN, Anna S.
    Objectives: Enterobacterales and other non-fermenting Gram-negative bacteria have become a threat worldwide owing to the frequency of multidrug resistance in these pathogens. On the other hand, efficacious therapeutic options are quickly diminishing. The aims of this study were to describe the susceptibility of 50 multiresistant Gram-negative bacteria, mostly pan-resistant, against old and less-used antimicrobial drugs and to investigate the presence of antimicrobial resistance genes. Methods: A total of 50 genetically distinct isolates were included in this study, including 14 Acinetobacter baumannii (belonging to ST79, ST317, ST835 and ST836), 1 Pseudomonas aeruginosa (ST245), 8 Serratia marcescens and 27 Klebsiella pneumoniae (belonging to STII, ST340, ST258, ST16, ST23, ST25, ST101, ST234, ST437 and ST442). The isolates were submitted to antimicrobial susceptibility testing and whole-genome sequencing to evaluate lineages and resistance genes. Results: Our results showed that some strains harboured carbapenemase genes, e.g. bla(K)(PC-)(2) (28/50; 56%) and bla(OXA-23) (11/50; 22%), and other resistance genes encoding aminoglycoside-modifying enzymes (49/50; 98%). Susceptibility rates to tigecycline (96%) in all species (except P. aeruginosa), to minocycline (100%) and doxycycline (93%) in A. baumannii, to ceftazidime/avibactam in S. marcescens (100%) and K. pneumoniae (96%), and to fosfomycin in S. marcescens (88%) were high. Chloramphenicol and quinolones (6% susceptibility each) did not perform well, making their use in an empirical scenario unlikely. Conclusions: This study involving genetically distinct bacteria showed promising results for tigecycline for all Gram-negative bacteria (except P. aeruginosa), and there was good activity of minocycline against A. baumannii, ceftazidime/avibactam against Enterobacterales, and fosfomycin against S. marcescens. (C) 2020 The Author(s).
  • article 6 Citação(ões) na Scopus
    Genetic and virulence characterization of colistin-resistant and colistin-sensitive A. baumannii clinical isolates
    (2019) LEITE, Gleice Cristina; STABLER, Richard A.; NEVES, Patricia; PERDIGAO NETO, Lauro V.; MARTINS, Roberta C. Ruedas; RIZEK, Camila; ROSSI, Flavia; LEVIN, Anna S.; COSTA, Silvia Figueiredo
    Treatment of infections caused by A. baumannii is becoming a challenge due to the ability to develop multidrug-resistance, virulence, and high mortality. We described the colistin resistance and virulence genes present in sixA. baumannii clinical isolates using WGS, expression by qPCR, and virulence in the Galleria mellonella model. The colistin-resistant isolates were assigned as ST233 and the colistin-susceptible isolates as ST236 and ST407. The colistin-resistant isolates contained mutations within PmrA/PmrB, and the pmrA showed up-regulation in all of them. Only one colistin-resistant isolate indicating virulence in G. mellonella. This particular isolate belonged to a different clone, and it was the only isolate that presented non-synonymous mutations in pmrB. Colistinresistance in A. baumannii isolates seems to be caused by up-regulation of pmrA gene. Only one isolate appeared to be virulent in the G. mellonella model. This finding indicating low virulence in isolates belonging to emerging clones circulating in our hospital.
  • article 7 Citação(ões) na Scopus
    Risk factors for bloodstream infection by multidrug-resistant organisms in critically ill patients in a reference trauma hospital
    (2022) CAMPOS, Luciana Rodrigues Pires de; CORTES, Marina Farrel; DEO, Beatriz; RIZEK, Camila; SANTOS, Sania; PERDIGAO, Lauro; COSTA, Silvia Figueiredo
    Background: Bloodstream infections (BSI) by multidrug-resistant (MDR) organisms are responsible for significant mortality in critically ill trauma patients. Our objective is to identify the risk factors for BSI by MDR agents and their resistance mechanisms in a trauma reference hospital. Methods: During 18 months, all patients admitted in our Intensive Care Unit (ICU) were enrolled in this prospective cohort. We included the first episode of BSI by carbapenem-resistant Gram-negative bacteria, methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant enterococcus. Demographic and clinical data were compared among patients with and without BSI and variables with P <.05 were tested in a multivariate analysis. We performed PCR for identification of carbapenemase and SCC mec genes and Pulsed-field gel electrophoresis for clonality. Results: Out of 1,528 patients, 302 (19.8%) were trauma and 66 (4.3%) had a MDR-BSI ( 19.5% were trauma). The multivariate analysis showed that mechanical ventilation (OR3.16; 95% CI 1-8; P =.02), hemodialysis (OR3.16; 95% CI 1-5; P =.0003) and surgery (OR1.76; 95% CI 1-3; P =.04) were independent risk factors for MDR- BSI. The most frequent MDR were Klebsiella pneumoniae (n = 26) and MRSA (n = 27). Regarding K pneumoniae strains (n = 24), 20 ( 83.8%) harbored bla KPC gene and 1 bla NDM. The majority of KPC isolates belonged to a predominant clone; while the MRSA were polyclonal and SCC mec type II. Conclusions: Mechanical ventilation, surgery and hemodialysis were independent risk factors for MDR-BSI in our cohort, but trauma was not. KPC was the main mechanism of resistance among carbapenem-resistant K pneumoniae that belonged to a predominant clone which could indicate cross-transmission.
  • article 17 Citação(ões) na Scopus
    Vancomycin-resistant enterococci isolates colonizing and infecting haematology patients: clonality, and virulence and resistance profile
    (2018) MARCHI, A. P.; PERDIGAO NETO, L. V.; MARTINS, R. C. R.; RIZEK, C. F.; CAMARGO, C. H.; MORENO, L. Z.; MORENO, A. M.; BATISTA, M. V.; BASQUEIRA, M. S.; ROSSI, F.; AMIGO, U.; GUIMARAES, T.; LEVIN, A. S.; COSTA, S. F.
    Background: Vancomycin-resistant enterococci (VRE) are an important agent of colonization and infection in haematology patients. However, the role of virulence on VRE colonization and infection is controversial. Aim: To characterize the lineage, virulence and resistance profile of VRE infection and colonization isolates; as well as their impact on outcome of haematology patients using a regression logistic model. Methods: Eighty-six isolates (80 Enterococcus faecium and six E. faecalis) from 76 patients were evaluated. Polymerase chain reaction for resistance and virulence genes, and pulsed-field gel electrophoresis and whole genome sequencing of the major clusters, were performed. Bivariate and multivariate analyses were carried out to evaluate the role of virulence genes on outcome. Findings: All isolates harboured the vanA gene. Regarding the virulence genes, 96.5% of isolates were positive for esp, 69.8% for gelE and asa1 genes. VRE infection isolates were more virulent than colonization isolates and harboured more often the gelE gene (P = 0.008). Infections caused by VRE carrying asal gene resulted more frequently in death (P = 0.004), but only the predominant clone remained as protector in the multivariate model. The E. faecium strains were assigned to seven STs (ST78, ST412, ST478, ST792, ST896, ST987, ST963) that belonged to CC17. The E. faecalis sequenced belonged to ST9 (CC9). Conclusion: E. faecium was predominant, and infection isolates were more virulent than colonization isolates and harboured more often the gene gelE. Infections caused by VRE carrying the asal gene appeared to be associated with a fatal outcome.
  • article 0 Citação(ões) na Scopus
    Conjugative transfer of plasmid p_8N_qac(MN687830.1) carrying qacA gene from Staphylococcus aureus to Escherichia coli C600: potential mechanism for spreading chlorhexidine resistance
    (2021) BES, Taniela Marli; NAGANO, Debora Satie; MARCHI, Ana Paula; CAMILO, Gaspar; PERDIGAO-NETO, Lauro Vieira; MARTINS, Roberta Ruedas; LEVIN, Anna Sara; COSTA, Silvia Figueiredo
    The methicillin resistant Staphylococcus aureus (MRSA) is recognized by its ability to acquire and transferring resistance genes through interspecies conjugative plasmids. However, transference of plasmids from Gram-positive cocci to Gram-negative bacilli is not well characterized. In this report, we describe the transfer of a conjugative plasmid carrying qacA from MRSA to Escherichia coli C600. We performed a conjugation experiment using a chlorhexidine resistant MRSA isolate (ST-105/SCCmec type III) carrying the gene qacA and qacC as the donor and a chlorhexidine susceptible E. coli C600 isolate as the receptor. Transconjugants were selected using MacConkey agar plates containing chlorhexidine in concentrations ranging from 0.25 to 16 g.L-1. To genotypically confirm the transfer of the resistance gene, the transconjugants were screened by Polymerase Chain Reaction (PCR) and submitted to Sanger's sequencing. MRSA isolates successfully transferred the chlorhexidine resistance gene (qacA) to the recipient E. coli strain C600. The E. coli transconjugant exhibited an important reduction of chlorhexidine susceptibility, with MICs increasing from <= 0.25 to >= 16 g.L-1 after conjugation. The qacA gene was detected by PCR as well as in the Sanger's sequencing analysis of DNA from transconjugant plasmids. To the best of our knowledge, this is the first report of the plasmid p_8N_qac(MN687830.1) carrying qacA and its transfer by conjugation from a MRSA to an E. coli. These findings increase concerns on the emergence of resistance dissemination across the genus and emphasizes the importance of continuous antiseptic stewardship.
  • article 4 Citação(ões) na Scopus
    Phenotypic and genotypic characteristics of a carbapenem-resistant Serratia marcescens cohort and outbreak: describing an opportunistic pathogen
    (2022) PRADO, Gladys; MENDES, Elisa Teixeira; MARTINS, Roberta Cristina Ruedas; PERDIGAO-NETO, Lauro Vieira; FREIRE, Maristela Pinheiro; MARCHI, Ana Paula; CORTES, Marina Farrel; LIMA, Victor Augusto Camarinha de Castro; ROSSI, Flavia; GUIMARAES, Thais; LEVIN, Anna Sara; COSTA, Silvia Figueiredo
    Serratia marcescens is an emerging opportunistic pathogen with high genetic diversity. This article describes the microbiological characteristics of isolates and the risk factors for infections caused by carbapenem-resistant S. marcescens. A retrospective study of patients colonized (n=43) and infected (n= 20) with carbapenem-resistant S. marcescens over a 3-year period was conducted. Polymerase chain reaction for carbapenemase genes and molecular typing of all available strains was performed. Forty-two isolates were analysed, including three environmental samples identified during an outbreak. Thirty-five carbapenem-resistant S. marcescens carried bla KPC-2, one isolate was bla(NDM)-positive and four isolates carried bla(OXA)-101. The genomes were grouped into three clusters with 100% bootstrap; three patterns of mutations on ompC and ompF were found. The strains carried virulence genes related to invasion and haemolysis, and the environmental strains presented fewer mutations on the virulence genes than the clinical strains. Multi-variate analysis showed that previous use of polymyxin (P= 0.008) was an independent risk factor for carbapenem-resistant S. marcescens infection. This study highlighted that bla KPC-2 in association with ompC or ompF mutation was the most common mechanism of resistance in the study hospital, and that previous use of polymyxin was an independent risk factor for carbapenem-resistant S. marcescens. There was a predominant clone, including the environmental isolates, suggesting that crosstransmission was involved in the dissemination of this pathogen.
  • article 6 Citação(ões) na Scopus
    Are mobile phones part of the chain of transmission of SARS- CoV-2 in hospital settings?
    (2021) ESPINOZA, Evelyn Patricia Sanchez; CORTES, Marina Farrel; NOGUERA, Saidy Vasconez; PAULA, Anderson Vicente de; GUIMARAES, Thais; BOAS, Lucy Santos Villas; PARK, Marcelo; SILVA, Cristina Carvalho da; MORALES, Ingra; NETO, Lauro Vieira Perdigao; TOZETTO-MENDOZA, Tania Regina; BOSZCZOWSKI, Icaro; SABINO, Ester Cerdeira; MENDES-CORREA, Maria Cassia; LEVIN, Anna Sara; COSTA, Silvia Figueiredo
    Mobile phones (MPs) have become an important work tool around the world including in hospitals. We evaluated whether SARS-CoV-2 can remain on the surface of MPs of first-line healthcare workers (HCW) and also the knowledge of HCWs about SARS-CoV-2 cross-transmission and conceptions on the virus survival on the MPs of HCWs. A crosssectional study was conducted in the COVID-19 Intensive Care Unit of a teaching hospital. An educational campaign was carried out on cross-transmission of SARS-CoV-2, and its permanence in fomites, in addition to the proper use and disinfection of MPs. Herewith an electronic questionnaire was applied including queried conceptions about hand hygiene and care with MP before and after the pandemic. The MPs were swabbed with a nylon FLOQ SwabTM, in an attempt to increase the recovery of SARS-CoV-2. All MP swab samples were subjected to SARS-CoV-2 RT-PCR; RT-PCR positive samples were subjected to viral culture in Vero cells (ATCC (R) CCL-81TM). Fifty-one MPs were swabbed and a questionnaire on hand hygiene and the use and disinfection of MP was applied after an educational campaign. Most HCWs increased adherence to hand hygiene and MP disinfection during the pandemic. Fifty-one MP swabs were collected and two were positive by RT-PCR (4%), with Cycle threshold (Ct ) values of 34-36, however, the cultures of these samples were negative. Although most HCWs believed in the importance of cross-transmission and increased adherence to hand hygiene and disinfection of MP during the pandemic, SARS-CoV-2 RNA was detected in MPs. Our results suggest the need for a universal policy in infection control guidelines on how to care for electronic devices in hospital settings.
  • article 8 Citação(ões) na Scopus
    Colistin-resistant Escherichia coli belonging to different sequence types: genetic characterization of isolates responsible for colonization, community- and healthcare-acquired infections
    (2021) PAIVA, Yrving; NAGANO, Debora Satie; COTIA, Andre Luis Franco; GUIMARAES, Thais; MARTINS, Roberta Cristina Ruedas; PERDIGAO NETO, Lauro Vieira; CORTES, Marina Farrel; MARCHI, Ana Paula; CORSCADDEN, Louise; MACHADO, Anna Silva; PAULA, Alexandre Inacio de; FRANCO, Lucas Augusto Moyses; NEVES, Patricia Regina; LEVIN, Anna Sara; COSTA, Silvia Figueiredo
    The plasmid-mediated colistin-resistance gene named mcr-1 has been recently described in different countries and it became a public health challenge. Of note, few studies have addressed the spread of Escherichia coli harboring the mcr-1 gene in both, community and hospital settings. A total of seven colistin-resistant E. coli carrying mcr-1, collected from 2016 to 2018, from community (n=4), healthcare-acquired infections (n=2) and colonization (n=1) were identified in three high complexity hospitals in Sao Paulo, Brazil. These colistin-resistant isolates were screened for mcr genes by PCR and all strains were submitted to Whole Genome Sequencing and the conjugation experiment. The seven strains belonged to seven distinct sequence types (ST744, ST131, ST69, ST48, ST354, ST57, ST10), and they differ regarding the resistance profiles. Transference of mcr-1 by conjugation to E. coli strain C600 was possible in five of the seven isolates. The mcr-1 gene was found in plasmid types IncX4 or IncI2. Three of the isolates have ESBL-encoding genes (bla(CTX-M-2), n=2; bla(CTX-M-8), n=1). We hereby report genetically distinct E. coli isolates, belonging to seven STs, harboring the mcr-1 gene, associated to community and healthcare-acquired infections, and colonization in patients from three hospitals in Sao Paulo. These findings point out for the potential spread of plasmid-mediated colistin-resistance mechanism in E. coli strains in Brazil.