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Introduction Enterobacteriaceae are often reported as a typical bacterial population in raw milk from any mammalian origin. The frequent concern with bacteria, especially those related to this group of microorganisms, is their increasing resistance to antibiotics and the emergence of enzymes that degrade them. This study aimed to characterize isolates of Enterobacteriaceae from raw goat milk to expose associated safety problems and possible technological challenges. Methods Isolates from 21 raw goat milk samples purchased in the State of Rio de Janeiro, Brazil, were identified by mass spectrometry, after isolation on Violet Red Bile Glucose agar. The isolates were subjected to evaluation of proteolytic, lipolytic, hemolytic, and biofilm producing activities. Furthermore, resistance profiles and production capacity of enzymes that degrade antimicrobials were evaluated. Results Almost half of the 59 isolates (48%) belonged to the Enterobacter genus, with a significant prevalence of the Serratia (20%) and Klebsiella (11%) genera. The majority showed biofilm-producing activity (90%), while the activity of degradative enzymes was observed in approximately 20%. Few isolates were found with a profile of resistance to antimicrobials, with only one isolate of Klebsiella variicola being classified as multidrug-resistant. However, chromogenic culture media showed high production of extended-spectrum beta-lactamases and carbapenemases (54% and 46%, respectively), as a presumptive identification. Conclusions A considerable degree of virulence was observed in the Enterobacteriaceae isolates, as well as the potential for undesirable technological damage. The characterization and identification of the isolates contributes to the improvement of the risk monitoring process of goat's milk. [ABSTRACT FROM AUTHOR]

This study aimed to analyze ESBL-producing Escherichia coli prevalence in urine samples collected between 2011-2019 in Curitiba, a large city in Brazil, and relating it to antibiotic consumption and sanitary conditions. This is a longitudinal study correlating prevalence of ESBL-producing E. coli isolates from urine samples with district-level antibiotic consumption and sociodemographic data during 2011-2019. E. coli isolates were tested for antibiotic susceptibility and ESBL by an automated method. Statistical analysis applied linear regressions, pooled ordinary least squares, and fixed effects models for districts or years. The Chow and Hausman tests indicated that the fixed effects model for individual districts fitted best. Chi-square test was used for qualitative variables (statistical significance was set when P < 0.05). Among the 886 535 urine sample cultures, 9.9% of isolates were ESBL-producing E. coli. Their prevalence increased from 4.7% in 2012 to 19.3% in 2019 (P < 0.0001; R2 = 0.922). This progressive increase correlated with age (P = 0.007; R2 = 0.8725) and male gender (P < 0.001) and increased antibiotic consumption (P = 0.0386; R2 = 0.47). The fixed effects model showed that district influences ESBL prevalence and that antibiotic consumption explains 20%-30% of this variation, with an increase of one defined daily dose accounting for an increase of 0.02084 percentage points of ESBL. The increasing prevalence of ESBL-producing E. coli can, to a considerable extent, be explained by increasing antibiotic consumption., (© The Author(s) 2024. Published by Oxford University Press on behalf of FEMS.)

Background: Bloodstream infections (BSI) are a global health issue, leading to high mortality and morbidity among hospitalized patients., Methods: A retrospective, observational and descriptive study was conducted by reviewing blood culture records collected from patients with suspected BSI, between January 2017 and December 2019., Results: The most frequent antimicrobial resistant (AMR) pathogens were methicillin-resistantStaphylococcus aureus(MRSA) (40%), methicillin-resistantS. epidermidis (MRSE) (9.5%), and extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae (35.3%)., Conclusions: Our findings underscore the importance of continued vigilance and advocate for the rational use of antimicrobial agents.

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 bla CTX-M and/or bla KPC 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. [ABSTRACT FROM AUTHOR]

Estended-spectrum beta-lactamases (ESBL) are bacterial enzymes that hydrolyses beta-lactam antibiotics with broad spectrum of action. Currently, ESBL-producing strains are important agents of nosocomial infection which may limit the available options available, necessitating the deployment of appropriate techniques for its detection. The ESBL is codified in plasmids that can be transferred to other microorganisms by conjugation. The indiscriminate use of broad-spectrum cephalosporins contributes to the selection of ESBL-producing strains and makes the problem of universal interest. This work carried out two techniques for detection of ESBL, the technique of Double Disks Diffusion and the technique of Clavulanate Disk Addition. Strains of Klebsiella sp (n=38) and Escherichia coli (n=114) routinely isolated from patients admitted in the university hospital from January 2004 to December 2005 were included in the study. Of those, 19 (50%) of samples of Klebsiella sp and 55 (48.3%) samples of E. coli were ESBL-producing. The technique the clavulanate disk addition showed greater sensitivity (100%) showing higher skill in the detection of ESBL, whereas the technique of Double Disk Diffusion had a sensitivity of 24.3%. Cefotaxima was the substrate that showed greater sensitivity in detecting ESBL. Both techniques showed high specificity (100%). [ABSTRACT FROM AUTHOR]

E. coli is the main pathogen of UTI. It is important to be aware the local epidemiological data for an appropriate initial treatment. Resistance to antimicrobial agents has increased, especially to first-choice antibiotics in the treatment of cystitis. There are few studies on the sensivity profile of community uropathogen in our region., Objective: To characterize antimicrobials the sensitivity profile to E. coli isolated from urocultures of women treated at Basic Health Units and Emergency Care Units of Londrina- Paraná- Brazil during a period of 12 months (June 1, 2016 to June 1, 2017)., Methodology: A cross-sectional study was carried out from June 2016 to June 2017. All urine samples collected in the Basic Health Units and Emergency Departments in the city of Londrina (Paraná State, Brazil) were sent to a Central Laboratory where the identification and antimicrobial susceptibility testing were performed. Clinical Laboratory Standards Institute (CLSI) breakpoints were used for the interpretation of susceptibility testing results., Results: 56,555 urine cultures were performed in the period, of which 8,832 were positive, of which 5,377 were women. Of these samples, 4.7% were enterobacteria producing extended-spectrum beta-lactamases (ESBL) and 15.5% resistant to quinolones. TMP- SMX was resistant in more than 30% of the samples in all age groups. Among quinolone-resistant isolates, resistance to cephalothin, ampicillin and sulfamethoxazole-trimethoprim was greater than 60%. Nitrofurantoin was the only antimicrobial that showed 90% of sensitivity., Conclusion: The antimicrobials sensitivity profile was similar to that reported in the literature, with TMP- SMX resistance greater than 30% in the studied samples. Nitrofurantoin maintains high sensitivity rates greater than 90%. Resistance to quinolones increases proportionally with age, as well ESBL., Competing Interests: Conflicts of interest The authors declare no conflicts of interest., (Copyright © 2022 Sociedade Brasileira de Infectologia. Published by Elsevier España, S.L.U. All rights reserved.)

Background: Carbapenem-resistant Acinetobacter baumannii (CRAB) is a growing threat to public health., Methods: A 3-year retrospective study was conducted to evaluate the prevalence and lethality of multidrug-resistant (MDR) A. baumannii isolated from Brazilian patients., Results: In this study, 219 Acinetobacter baumannii isolates were identified, of which 70.8% (155/219) were isolated from patients hospitalized in intensive care units. Of these, 57.4% (n = 89/155) were assessed, of which 92.1% (82/89) were carbapenem-resistant, and 49 were classified as infected. The lethality rate was 79.6% (39/49)., Conclusions: We highlight the need of an effective epidemiological surveillance measure to contain the dissemination of CRAB in the hospital environment.

Antimicrobial resistance is among the most serious public health threats of the 21st century, with great impact in terms of One Health. Among antimicrobial resistant bacteria (ARB), extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli (ESBL-EC) represent major challenges to human healthcare. Wild birds have been commonly used as environmental bioindicators of ESBL-EC. Remote locations represent a unique opportunity to evaluate the occurrence, dissemination and epidemiology of ARB in the environment. Herein we surveyed ESBL-EC in 204 cloacal swabs from six nonsynanthropic seabird species at the pristine Rocas Atoll, Brazil. We identified ESBL-EC isolates in 2.4% (5/204) of the tested seabirds, all in magnificent frigatebirds (Fregata magnificens). We isolated strains of O25b-ST131-fimH22 harboring gene bla CTX-M-8 (3 clones), ST117 harboring gene bla SHV-12 , and a novel ST11350 (clonal complex 349) harboring genes bla CTX-M-55 and fosA3. All the isolates presented Extraintestinal pathogenic E. coli (ExPEC) virulence profiles. We suggest that magnificent frigatebirds may act as "flying bridges", transporting ESBL-EC and ARGs from an anthropogenically-impacted archipelago geographically close to our pristine and remote study site. The characteristics of our isolates suggest zoonotic potential and, despite the apparent good health of all the evaluated birds, may represent a hypothetical potential threat to the avian population using the atoll. To our knowledge, this is the first description of: (1) the pandemic and public health relevant ST131-O25b harboring bla CTX-M-8 worldwide; (2) ST131-fimH22 in wild birds; and (3); fosA3 in wildlife. Our findings expand the current epidemiological knowledge regarding host and geographical distribution of ESBL-EC and ARGs in wild birds, and emphasize the disseminating characteristics and adaptability of ST131 and ST117 strains within the human-animal-interface. Herein we discuss the involvement of nonsynanthropic wild birds in the epidemiology of antimicrobial resistance and their potential as sentinels of ESBL E. coli in insular environments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

VIM, IMP, and NDM carbapenemases are metallo-β-lactamases (MBLs) that are widely distributed throughout the world. SPM-1 (Sao Paulo metallo-β-lactamase) is an MBL that was described in Pseudomonas aeruginosa in Sao Paulo (Brazil) in 2002. We report for the first time the presence of SPM-1 in Chile, in an isolate of P aeruginosa resistant to meropenem and imipenem, detected in a carbapenemase surveillance rectal swab culture, in a patient admitted to our institution. The sequence of the PCR product was 100% identical to the sequence of SPM-1 reported in Brazil. The patient had a history of an angioplasty performed in Brazil in 2004-2005. As a consequence of this finding, the detection of SPM by PCR will be incorporated into the routine screening for carbapenemases in P aeruginosa.

Pseudomonas aeruginosa is associated with chronic and progressive lung disease and is closely related to increased morbidity and mortality in cystic fibrosis (CF) patients. Hypermutable (HPM) P. aeruginosa isolates have been described in these patients and are usually associated with antibiotic resistance. This study aimed to investigate the occurrence of carbapenem resistance and hypermutable phenotype in 179 P. aeruginosa isolates from 8 chronically CF patients assisted at two reference centers in Rio de Janeiro, Brazil. Using disk diffusion test, non-susceptible (NS) rates higher than 40% were observed for imipenem, amikacin, and gentamicin. A total of 79 isolates (44.1%), 71 (39.6%), and 8 (4.4%) were classified as carbapenem-resistant (CR resistance to at least one carbapenem), multidrug-resistant (MDR), and extensively drug-resistant (XDR), respectively. Minimal inhibitory concentration was determined for 79 CR P. aeruginosa and showed the following variations: 4 and 128 μg/mL to imipenem, 4 and 64 µg/mL to meropenem, and 4 and ≥ 32 µg/mL to doripenem. We have found only four (2.23%) HPM isolates from 4 patients. Analyzing the genetic relationship among the HPM isolates, 3 pulsed-field gel electrophoresis/pulsotypes (D, M, and J) were observed. Only M pulsotype was recovered from two patients in different years. Polymerase chain reaction screening for bla GES , bla IMP , bla KPC , bla NDM , bla OXA-48 , bla SPM , and bla VIM genes was performed for all CR isolates and none of them were positive. Our results demonstrate a high occurrence of CR and MDR P. aeruginosa of CF patients follow-up in both centers studied, while the presence of HPM is still unusual.

The occurrence of antibiotics in the natural environment has been a growing issue and correlations between this presence and developing resistance bacteria are explored. The purpose of this study was to investigate the presence of antibiotics of different classes and associated resistant bacteria, in water samples taken from urban river waters in Curitiba, Brazil. A method for the quantification of antibiotics (azithromycin, amoxicillin, norfloxacin ciprofloxacin, doxycycline and sulfamethoxazole) was developed and validated using liquid chromatography coupled with mass spectrometry. To investigate and identify coliforms resistant to these antibiotics, we performed selective microbiological culturing techniques. We detected antibiotics in our water samples; concentrations ranged from 0.13 to 4.63 μg L -1 , with the highest being amoxicillin at 4.63 μg L -1. In all water samples this study, antibiotic resistant bacteria were detected. Escherichia coli was resistant to amoxicillin, norfloxacin, ciprofloxacin, doxycycline and sulfamethoxazole. Strains producing β-lactamase with extended spectrum (ESBL and AmpC) were also found in these isolates. Enterococcus spp. displayed resistance to norfloxacin and ciprofloxacin, and some isolates were resistant to vancomycin, gentamicin and streptomycin (complementary tests). No P. aeruginosa resistant strains were observed. It is possible these antibiotics came from domestic effluents and may be contributing to the spread of bacterial resistance., (Copyright © 2020 Elsevier B.V. All rights reserved.)

After the sudden death of captive marmosets in São Paulo, Brazil, we conducted a histologic and microbiologic study. We found hyperacute septicemia caused by hypermucoviscous sequence type 86 K2 Klebsiella pneumoniae. We implemented prophylactic antimicrobial therapy, selected dedicated staff for marmoset interactions, and sanitized the animals' fruit to successfully control this outbreak.

Coastal areas with important economic activities have high levels of contamination by metals, pathogenic bacteria, among other contaminants. The emergence of antibiotic-resistant bacteria is a global problem of public health. Carbapenem resistant Enterobacteriaceae (CRE) are a serious threat. The occurrence of carbapenem resistant bacteria was investigated in waters and sediments of a Brazilian coastal area, characterized by high levels of contamination. The samples of water and sediment were collected in two areas of the coast of São Paulo (Brazil). The study involved the characterization of the molecular mechanisms associated with the carbapenem resistance phenotype. No genes were detected for β-lactamases but the absence and/or presence of mutations in outer membrane proteins (OMPs) may justify the detected phenotype. The presented results show the need for further studies that allow a review of the current legislation and the importance of the reevaluation of monitoring policies of these environments., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Objectives: Urinary tract infections (UTIs) caused by multidrug-resistant Escherichia coli have become a major medical concern. Old antibiotics such as fosfomycin have become an alternative therapeutic option due to their effectiveness and, as a result, fosfomycin is now used as a first-line drug for the treatment of UTIs in many countries. Despite low resistance rates, fosfomycin heteroresistance, defined as a phenomenon where subpopulations of bacteria are resistant to high antibiotic concentrations whereas most of the bacteria are susceptible, is an underestimated problem., Methods: The frequency of heteroresistance in E. coli isolated from hospitalized patients in Brazil and its effect on susceptibility of E. coli in biofilms was studied and the isolates were molecularly characterized to reveal the mechanisms behind their fosfomycin heteroresistance using whole-genome sequencing., Results: A higher frequency of fosfomycin heteroresistance compared with other studies was found. In biofilms, most heteroresistant isolates were less sensitive to fosfomycin than control isolates and showed overexpression of metabolic genes thereby increasing their survival rate. Molecular characterization showed that some resistant subpopulations derived from heteroresistant isolates had a defect in their fosfomycin uptake system caused by mutations in transporter and regulatory genes, whereas others overexpressed the murA gene. None to minor effects on bacterial fitness were observed. Oxidative stress protection, virulence and metabolic genes were differentially expressed in resistant subpopulations and heteroresistant isolates., Conclusion: Frequent detection of heteroresistance in UTIs may play a role in the failure of antibiotic treatments and should therefore be more carefully diagnosed., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Acinetobacter baumannii is an emerging pathogen associated with nosocomial infections and multidrug resistance. Polymyxin B has been used to treat infections caused by multidrug-resistant (MDR) A. baumannii but an increase in polymyxin B resistance has been observed. We aimed to determine the diversity, antimicrobial susceptibility, presence of polymyxin B heteroresistance and adaptive resistance in 72 A. baumannii clinical isolates from two public hospitals in Rio de Janeiro. The isolates were identified by sequencing of rpoB gene. Determination of the genetic diversity of isolates was performed by pulsed-field gel electrophoresis and oxacillinases genes were detected by polymerase chain reaction. The polymyxin B heteroresistance was analyzed by population analysis profile and adaptive resistance was evaluated after serial daily passages of isolates in broth containing increasing polymyxin B concentrations. The results showed that 49% of the isolates were collected from respiratory system and 62% were MDR, while 35% were extensively drug resistant. Additionally, all the isolates carried bla OXA-23-like , bla OXA-51-like genes and ISAba1, while 1% had bla OXA-24-like gene. The association of ISAba1-bla OXA-23 was found in 96% of the isolates. Polymyxin B heteroresistance was found in 36% of the isolates and polymyxin B adaptive resistance was not found in the isolates. Our study demonstrated the high resistance to antimicrobials used in clinical practice and the spread of oxacillinases genes and insertion sequence (IS). We also reported the presence of heteroresistance to polymyxin B used as a last-resort therapy for MDR A. baumannii.

Carbapenem resistance in Acinetobacter baumannii is a public health issue globally, mainly due to the production of carbapenem hydrolyzing class D β-lactamases (CHDLs). In Brazil, OXA-23 and OXA-143 CHDLs have been prevalent in A. baumannii from clinical settings, with some OXA-23 reports in the environmental samples, whereas OXA-72 has begun to be increasingly reported. This study aims to perform the genomic and microbiological characterization of carbapenem-resistant A. baumannii isolates recovered from migratory birds and captive birds inhabiting a lake within a Brazilian Zoo. Four hundred and eighty-one gram-negative bacilli were recovered from choanal and cloacal swabs obtained from 50 migratory birds and 37 captive birds present at the zoo's lake between July and August of 2012. Among all GNB, nine OXA-72-producing A. baumannii were detected from the microbiota of four migratory and five captive aquatic birds. The OXA-72-producing A. baumannii isolates were submitted to antimicrobial susceptibility test and PFGE, exhibiting a multidrug-resistant profile and clonal relatedness with OXA-72-positive human isolates circulating for eighteen years in a hospital setting. MLST, plasmid analysis and whole-genome sequencing revealed which all carbapenem-resistant A. baumannii from bird and human hosts belonged to clonal complex 79, and harboured a small plasmid (⁓16.6-kb in size), named pAC1-BRL, which carried bla OXA-72 gene, macrolide resistance genes msrE and mphE, and the toxin-antitoxin system AbkAB. To determine the impact of pAC1-BRL acquisition in the the capacity of a microorganism to survive in a competitive environment (in the following called fitness), the laboratory strain A. baumannii ATCC 19606 was used in the fitness experiments and suggested an increase of its relative fitness after the pAC1-BRL acquisition. In summary, the detection of OXA-72-producing A. baumannii strains belonging to CC79 in aquatic birds is a piece of epidemiological evidence demonstrating that dissemination of high-risk bacteria is extending beyond the hospital., Competing Interests: Declaration of competing interest A.C.G. has recently received research funding and/or consultation fees from Bayer, Eurofarma, MSD, Pfizer, and Zambon. Other authors have nothing to declare., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objectives: Carbapenemase-producing Enterobacterales are frequently involved in healthcare-associated infections worldwide. The objectives of this study were to investigate (i) the frequency of the main genes encoding carbapenemases, 16S rRNA methylases and aminoglycoside-modifying enzymes (AMEs) as well as the mcr gene and (ii) the clonal relationship of enterobacteria isolates resistant to carbapenems and aminoglycosides from colonisation and infection in patients from hospitals in northeastern Brazil., Methods: Antimicrobial susceptibility was determined using an automated VITEK®2 system. Presence of carbapenemase, AME and 16S rRNA methylase genes as well as the mcr gene was determined by PCR and amplicon sequencing. Genetic variability was determined by ERIC-PCR., Results: A total of 35 isolates resistant to carbapenems and aminoglycosides were selected for this study. Klebsiella pneumoniae was most common (45.7%), followed by Proteus mirabilis (28.6%) and Serratia marcescens (25.7%). AME genes were found in 97.1% of isolates, most commonly aph(3')-VI and aac(6')-Ib. The bla NDM-1 and bla KPC-2 genes were detected in 25.7% and 88.6% of isolates, respectively; five isolates harboured these genes concomitantly. According to the literature, this is the first report of the association of bla NDM-1 and bla KPC-2 in P. mirabilis and S. marcescens in Brazil. The isolates showed a multiclonal profile by ERIC-PCR., Conclusion: The emergence of bla NDM-1 associated with bla KPC-2 and AME genes in K. pneumoniae, P. mirabilis and S. marcescens isolates with a multiclonal profile is of concern as this limits therapeutic options. These results should alert medical authorities to establish rigorous detection methods to reduce the spread of these antimicrobial resistance genes., (Copyright © 2019 International Society for Antimicrobial Chemotherapy. Published by Elsevier Ltd. All rights reserved.)

The prevalence of carbapenem-resistant Pseudomonas aeruginosa is increasing. Identification of carbapenemase-producing P. aeruginosa will have therapeutic, epidemiological, and infection control implications. This study evaluated the performance of the EDTA-modified carbapenem inactivation method (eCIM) in tandem with the modified carbapenem inactivation method (mCIM) against a large collection of clinical P. aeruginosa isolates ( n  = 103) to provide clinicians a phenotypic test that not only identifies carbapenemase production but also distinguishes between metallo-β-lactamase and serine-carbapenemase production in P. aeruginosa The mCIM test was performed according to Clinical and Laboratory Standards Institute guidelines, while the eCIM was conducted as previously described for Enterobacteriaceae Test performance was compared to the genotypic profile as the reference. mCIM testing successfully categorized 91% (112/123) of P. aeruginosa isolates as carbapenemases or non-carbapenemase producers, with discordant isolates being primarily Guiana extended-spectrum (GES)-type producers. To increase the sensitivity of the mCIM for GES-harboring isolates, a double inoculum, prolonged incubation, or both was evaluated, with each modification improving sensitivity to 100% (12/12). Upon eCIM testing, all Verona integrin-encoded metallo-β-lactamases (VIM; n  = 27) and New Delhi metallo-β-lactamases (NDM; n  = 13) tested had 100% concordance to their genotypic profiles, whereas all Klebsiella pneumoniae carbapenemase (KPC; n  = 8) and GES ( n  = 12) isolates tested negative, as expected, in the presence of EDTA. The eCIM failed to identify all imipenemase (IMP)-producing ( n  = 22) and Sao Paulo metallo-β-lactamase (SPM)-producing ( n  = 14) isolates. KPC-, VIM-, and NDM-producing P. aeruginosa were well defined by the conventional mCIM and eCIM testing methods; additional modifications appear required to differentiate GES-, IMP-, and SPM-producing isolates., (Copyright © 2020 American Society for Microbiology.)

Multidrug resistance mediated by β-lactamase in Gram-negative bacilli is a serious public health problem. Sewers are considered reservoirs of multiresistant bacteria due to presence of antibiotics that select them and favor their dissemination. The present study evaluated the antibiotic resistance profile and β-lactamases production in Gram-negative bacilli isolates from hospital sewage and urban wastewater treatment plants (UWWTP) in Brazil. Bacteria were isolated and identified with biochemical tests. Antibiotic susceptibility testing was performed by the disk-diffusion method and detection of extended-spectrum β-lactamase and carbapenemases by enzymatic inhibitor and conventional PCR. Differences in resistance to amoxicillin clavulanic, aztreonam, cefepime, and cefotaxime were observed in hospital sewage compared with urban sewage (p < 0.05). The multidrug-resistant phenotype was observed in 33.3% of hospital sewage isolates (p = 0.0025). β-lactamases genes were found in 35.6% of isolates, with the most frequent being bla KPC and bla TEM (17.8%), and bla SHV and bla CTX-M (13.3% and 8.9%, respectively). The data obtained are relevant, since the bacteria detected are on the priority pathogens list from the World Health Organization and hospital sewage could be released untreated into the municipal collection system, which may favor the spread of resistance. Changes in hospital sewage discharge practices, as well as additional technologies regarding effluent disinfection in the UWWTP, can prevent the spread of these bacteria into the environment and negative impact on water resources.

Infections due to multidrug resistant Gram-negative pathogens are of great concern worldwide, as they are frequently associated with high mortality and morbidity rates. The occurrence of Pseudomonas spp. producing Klebsiella pneumoniae carbapenemases (KPCs) imposes a great challenge through treatment course of bloodstream infections (BSIs). Pseudomonas putida has been recognized as an emerging pathogen of healthcare associated infections (HAIs). Therefore, we aimed to report a case of a non-fatal case of peripheral line associated BSI (PLA-BSI) in an immunocompromised host due to P. putida harboring blaKPC-2 gene in Brazil. A P. putida isolate was recovered from a blood culture of a 72-year-old man admitted at a University Hospital, identified by BD Phoenix™ 100 (Becton, Dickinson and Company), causing PLA-BSI. The species identification was confirmed by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) and resistance to carbapenems were confirmed by Epsilometer test (E-test®). Additionally, the presence of important carbapenemases genes (blaKPC, blaNDM, blaOXA-48-like, blaSPM, blaIMP, blaVIM) was investigated by Polymerase Chain Reaction. The bacterial isolate was confirmed as meropenem resistant P. putida harboring blaKPC-2 gene.Thereofre, these fidings suggest that P. putida can work as a reservoir for resistance genes as this bacterium has the ability to disseminate through water-fluids inside hospital and community settings. Moreover, this paper highlights that a frequent and worldwide disseminated mechanism of resistance (blaKPC-2) is currently occurring among uncommon agents of BSI., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2020 Raiane Cardoso Chamon, Jaqueline Abel da Rocha, Isabella Araujo Martins, Laís Lopes Pires, Breno Macedo de Almeida, Nahara Souza Leite, Claudia Rezende Vieira de Mendonca Souza, Viviane Zahner, Rachel Leite Ribeiro, Thiago Pavoni Gomes Chagas, Ianick Souto Martins.)

The objective of this study was to verify the presence of antimicrobial resistant strains of Escherichia coli in pig farms and to use it as a biomarker to evaluate phenotypic and genotypic profiles of antimicrobial susceptibility, as well as the presence of Extended Spectrum Beta-lactamases (ESBLs) and fluoroquinolone resistance genes. Several samples (n = 306) collected from swine farms (n = 100) of Southern Brazil were used for E. coli isolation: 103 of swine feces, 105 of water, and 98 of soil. E. coli isolates were submitted to the disk-diffusion test to verify their antimicrobial susceptibility, to disk-approximation test to detect ESBL-producers, and to PCR analysis to search for ESBLs genes (blaCTY-M2, blaSHV-1, blaTEM-1, blaCTX-M2, blaOXA-1, blaPSE-1) and quinolone resistance genes (qnrA, qnrB and qnrS). The percentage of E. coli isolates found in feces, water and soil samples was 66.02%, 30.48% and 35.71%, respectively. The highest percentages of resistance were obtained for sulfamethoxazole associated with trimethoprim (63.70%), colistin (45.19%) and enrofloxacin (39.26%). Regarding the levels of multidrug resistance, 37.04% of the isolates were resistant to three or more classes of antimicrobials. The most common profile (16%) of multirresistance was GEM-SUT-ENO-COL. The index of multiple resistance to antimicrobials (IRMA) was above 0.2 in 78% of the multiresistant isolates. Out of 135 E. coli isolates, 7.41% was ESBL-producers, of which 50% showed the blaCMY-M2 gene, 40% the blaTEM-1 and 70% the qnrS gene. Of non-ESBL-producing strains resistant to enrofloxacin, 13.04% were positives for qnrS gene. These results demonstrated the presence of fecal contamination in the environment, in addition to high resistance indexes for several antimicrobials, including beta-lactams and fluoroquinolones, which was confirmed by the genetic detection of ESBLs and qnr genes., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The AmpC enzyme is normally expressed constitutively in Escherichia coli, and its overproduction confers resistance to cefoxitin. A newly reported AmpC, the extended-spectrum AmpC (ESAC), is related to resistance to cefepime, a fourth-generation cephalosporin. This enzyme presents more flexibility in the active site due to insertions, replacements, and deletions on AA sequences. Many isolates producing ESAC were reported in human clinical isolates, but E. coli ESAC producers were reported in animals only in France. The animal E. coli strains can produce this enzyme and possibly disseminate it to human and production environments. In our study, 3 strains of E. coli from milk and feces bovine samples, collected in Rio de Janeiro, Brazil, were suspected to produce ESAC. After excluding other mechanisms of resistance, the gene was sequenced to verify ESAC characteristics. These strains presented replacement of AA in omega and R2 loops, suggesting ESAC production. This is the first report to study ESAC E. coli in dairy farms in Brazil., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

In Brazil, the presence of plasmid-mediated AmpC (pAmpC)-producing isolates has been sporadically reported. We evaluated the frequency of pAmpC among 133 Enterobacteriaceae clinical isolates. The bla CMY-2-like gene was detected in a single Klebsiella pneumoniae isolate. In our study, the pAmpC frequency was very low as previously reported.

An outbreak of extended spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBLKp) infections in a neonatal intensive care unit (NICU) prompted a prospective investigation of colonization and infection with this pathogen. From August 1, 1997 to May 30, 1999, neonates admitted to the NICU for more than 24 h were screened for ESBLKp acquisition. Neonatal gastrointestinal screening was performed by means of faecal sampling within 48 h of admission and then weekly until discharge. Isolates were typed using pulsed-field gel electrophoresis (PFGE). Time-dependent proportional hazard models were used to identify independent effects of invasive procedures and antimicrobials after controlling for duration of stay at the NICU. During the study period, 464 neonates were admitted and 383 were regularly screened. Infections occurred in 13 (3.4%) neonates and 206 (53.8%) became colonized. Independent risk factors for colonization during the first nine days in the NICU were the antimicrobial combination cephalosporin plus aminoglycoside [hazard rate (HR)=4.60; 95% CI: 1.48-14.31], and each NICU-day was associated with a 26% increase in the hazard rate for colonization (HR=1.26; 95% CI: 1.16-1.37). Previous colonization (HR=5.19; 95% CI: 1.58-17.08) and central vascular catheter use (HR=13.89; 95% CI: 2.71-71.3) were independent risk factors for infection. In an outbreak setting the proportion of neonates colonized with ESBLKp was observed to increase with the duration of stay and antimicrobial use, and once colonized, infants exposed to invasive devices may become infected.