Genetic diversity of KPC-2-producing Klebsiella pneumoniae complex from aquatic ecosystems.

During the COVID-19 pandemic, the occurrence of carbapenem-resistant Klebsiella pneumoniae increased in human clinical settings worldwide. Impacted by this increase, international high-risk clones harboring carbapenemase-encoding genes have been circulating in different sources, including the environment. The blaKPC gene is the most commonly disseminated carbapenemase-encoding gene worldwide, whose transmission is carried out by different mobile genetic elements. In this study, blaKPC-2-positive Klebsiella pneumoniae complex strains were isolated from different anthropogenically affected aquatic ecosystems and characterized using phenotypic, molecular, and genomic methods. K. pneumoniae complex strains exhibited multidrug-resistant and extensively drug-resistant profiles, spotlighting the resistance to carbapenems, ceftazidime-avibactam, colistin, and tigecycline, which are recognized as last-line antimicrobial treatment options. Molecular analysis showed the presence of several antimicrobial resistance, virulence, and metal tolerance genes. In-depth analysis showed that the blaKPC-2 gene was associated with three different Tn4401 isoforms (i.e., Tn4401a, Tn4401b, and Tn4401i) and NTEKPC elements. Different plasmid replicons were detected and a conjugative IncN-pST15 plasmid harboring the blaKPC-2 gene associated with Tn4401i was highlighted. K. pneumoniae complex strains belonging to international high-risk (e.g., ST11 and ST340) and unusual clones (e.g., ST323, ST526, and ST4216) previously linked to clinical settings. In this context, some clones were reported for the first time in the environmental sector. Therefore, these findings evidence the occurrence of carbapenemase-producing K. pneumoniae complex strains in aquatic ecosystems and contribute to the monitoring of carbapenem resistance worldwide.

Klebsiella quasipneumoniae subsp. similipneumoniae ST1859 O5:KL35 from Soil: First Report of qnrE1 in the Environment.

A Klebsiella quasipneumoniae subsp. similipneumoniae strain, named S915, belonging to the ST1859 O5:KL35, and harboring the plasmid-mediated quinolone resistance qnrE1 gene, was isolated from a soil sample cultivated with lettuce in Brazil. The core genome multilocus sequence typing analysis revealed that S915 strain was most related to a clinical strain of Brazil. Comparative genomic analysis showed that ST1859 O5:KL35 strains have been circulating in clinical settings and are closely related to multidrug resistance and multimetal tolerance. Strain S915 presented a plasmid contig co-harboring the qnrE1 gene and tellurite tolerance operon. The region harboring the qnrE1 gene (ISEcp1-qnrE1-araJ-ahp) shared high similarity with others from infected humans, ready-to-eat dish, and food-producing animals in Brazil. This is the first report of the plasmid-mediated qnrE1 gene in the environment. Our findings evidence the initial dissemination of the qnrE1 gene in the environment by the introduction of a clinical strain, which may be spread to different sectors, representing a One Health challenge.

Patterns of antimicrobial resistance and metal tolerance in environmental Pseudomonas aeruginosa isolates and the genomic characterization of the rare O6/ST900 clone.

Pseudomonas aeruginosa can harbor several virulence and antimicrobial resistance genes (ARGs). In this regard, virulent and multidrug-resistant (MDR) P. aeruginosa strains are closely related to severe infections. In addition, this species can also carry metal tolerance genes, selecting mainly antimicrobial-resistant strains. The action of several pollutants on the environment may favor the occurrence of antimicrobial-resistant and metal-tolerant strains. Therefore, the aim of this study was to characterize potentially pathogenic, antimicrobial-resistant, and/or metal-tolerant P. aeruginosa isolates from different environmental samples (waters, soils, sediments, or sands) and to perform a whole-genome sequence-based analysis of a rare clone from residual water. Environmental isolates carried virulence genes related to adherence, invasion, and toxin production, and 79% of the isolates harbored at least five virulence genes. In addition, the isolates were resistant to different antimicrobials, including important antipseudomonal agents, and 51% of them were classified as MDR, but only ARGs associated with aminoglycoside resistance were found. Furthermore, some isolates were tolerant mainly to copper, cadmium, and zinc, and presented metal tolerance genes related to these compounds. Whole-genome characterization of an isolate with unique phenotype with simultaneous resistance to antimicrobials and metals showed nonsynonymous mutations in different antimicrobial resistance determinants and revealed a classification of O6/ST900 clone as rare, potentially pathogenic, and predisposed to acquire multidrug resistance genes. Therefore, these results draw attention to the dissemination of potentially pathogenic, antimicrobial-resistant, and metal-tolerant P. aeruginosa isolates in environmental niches, alerting to a potential risk mainly to human health.

Characterization of the virulence, agr typing and antimicrobial resistance profile of Staphylococcus aureus strains isolated from food handlers in Brazil.

Staphylococcus aureus is one of the main pathogens associated with foodborne outbreaks in Brazil and food handlers can carry toxigenic and resistant S. aureus strains. The aims of this study were to verify the frequency of virulence genes, to identify the agr groups and to determine the antimicrobial resistance profile of S. aureus strains isolated from food handlers of pilot kitchens located in São Paulo, Brazil. A total of 74 strains of the Staphylococcus genus were detected and 50% were identified as of the species S. aureus. The enterotoxin genes detection, tst and luk-PV detection, agr typing, mecA detection, ccr complex detection and SCCmec typing were performed using PCR. The antimicrobial resistance testing was performed by the disk diffusion method. The enterotoxin genes were identified in 36 S. aureus, including sea (83.8%). The tst gene was detected in 18.92% of the strains and the luk-PV was detected in only one isolate. Agr typing classified 58.3% of the strains as type I. Seven (18.92%) strains were classified as MRSA and the ccr2 complex was detected in six of these isolates. The SCCmec typing characterized strains as type II, III, IV and V. Moreover, there were also a greater number of resistant strains to penicillin (83.78%) and clarithromycin (67.57%). In conclusion, the study revealed a significant prevalence of S. aureus, and the presence of different virulence genes and a worrying resistance profile in S. aureus strains isolated from food handlers in this country.

Occurrence and genetic characteristics of multidrug-resistant Escherichia coli isolates co-harboring antimicrobial resistance genes and metal tolerance genes in aquatic ecosystems.

Multidrug-resistant (MDR) Escherichia coli isolates (n = 50) were recovered from aquatic ecosystems, which presented high counts of E. coli and metal values within the recommended range. These isolates showed different multidrug resistance profiles, highlighting the resistance to extended-spectrum cephalosporins, polymyxins, and fluoroquinolones. Several antimicrobial resistance genes (ARGs) were found, spotlighting the presence of at least one ß-lactamase-encoding gene in each E. coli isolate. Substitutions in the quinolone resistance-determining regions and the two-component systems involving PhoP/PhoQ and PmrA/PmrB were also found. The metal tolerance gene rcnA (nickel and cobalt efflux pump) was the most prevalent. In this regard, 94% of E. coli isolates presented the co-occurrence of at least one ARG and metal tolerance gene. Furthermore, virulence genes and genetic diversity were found among MDR E. coli isolates. The emergence of potentially pathogenic isolates exhibiting multidrug resistance and metal tolerance emerged as a global health problem at the human-animal-environment interface.

Quercetin-loaded chitosan nanoparticles as an alternative for controlling bacterial adhesion to urethral catheter.

OBJECTIVES: The present study aimed to assess the antimicrobial and antiadhesion behavior of quercetin-loaded chitosan nanoparticles in Escherichia coli and Staphylococcus aureus multidrug-resistant isolates. METHODS: The ionic gelation method was used to prepare chitosan nanoparticles loaded with quercetin. The antimicrobial and antibiofilm effects were observed by minimum inhibitory concentration (MIC), plate count, crystal violet assay, and the matrix exopolysaccharide dosages. The nanoparticles coated in silicone urethral catheters were evaluated by crystal violet assay and plating count method. RESULTS: MIC ranged from 6.25 to 12.5 mg/ml. A reduction of at least 3.6 log CFU/ml and 6.2 log CFU/ml for, respectively, E. coli and S. aureus isolates was observed (p < 0.05). Under subinhibitory concentration (3.1 mg/ml) it was found a reduction of microbial adhesion and exopolysaccharide dosages in respectively 83.3% and 75% of the bacterial samples. The coated silicone urethral catheters showed a reduction of adhered cells in 25% of the isolates and biomass decreasing in 91.6% of them (p < 0.05). CONCLUSIONS: The quercetin nanoparticles provided antimicrobial and antiadhesion effects in multidrug-resistant isolates.

Characterization of the virulence, agr typing and antimicrobial resistance profile of Staphylococcus aureus strains isolated from food handlers in Brazil

Abstract Staphylococcus aureus is one of the main pathogens associated with foodborne outbreaks in Brazil and food handlers can carry toxigenic and resistant S. aureus strains. The aims of this study were to verify the frequency of virulence genes, to identify the agr groups and to determine the antimicrobial resistance profile of S. aureus strains isolated from food handlers of pilot kitchens located in São Paulo, Brazil. A total of 74 strains of the Staphylococcus genus were detected and 50% were identified as of the species S. aureus. The enterotoxin genes detection, tst and luk-PV detection, agr typing, mecA detection, ccr complex detection and SCCmec typing were performed using PCR. The antimicrobial resistance testing was performed by the disk diffusion method. The enterotoxin genes were identified in 36 S. aureus, including sea (83.8%). The tst gene was detected in 18.92% of the strains and the luk-PV was detected in only one isolate. Agr typing classified 58.3% of the strains as type I. Seven (18.92%) strains were classified as MRSA and the ccr2 complex was detected in six of these isolates. The SCCmec typing characterized strains as type II, III, IV and V. Moreover, there were also a greater number of resistant strains to penicillin (83.78%) and clarithromycin (67.57%). In conclusion, the study revealed a significant prevalence of S. aureus, and the presence of different virulence genes and a worrying resistance profile in S. aureus strains isolated from food handlers in this country.

Main aspects and prophylactic measures of COVID-19 in Brazil / Principais aspectos e medidas profiláticas da COVID-19 no Brasil

From the first case of COVID-19 in Brazil, the country became the third in the world in the raking of cases and deaths. Despite the measures implemented by the government, the number of infected and killed by COVID-19 continues to increase and the country faces several other problems that include social and political aspects, making it difficult to contain the pandemic. The present study addressed the general characteristics of SARS-CoV-2, pointed out the main socio-epidemiological aspects in Brazil and the treatment of COVID 19. A literature review was carried out to search for articles in PubMed, Scielo and Google Scholar databases. Patients with COVID-19 may be asymptomatic, but among symptomatic patients, the severity of the disease is related to age and pre-existing medical conditions. The lungs are the organs most affected by the virus and, for this reason, respiratory manifestations such as cough, shortness of breath, sputum production, sore throat and nasal congestion are the symptoms most associated with COVID-19. The transmission of SARSCoV-2 between humans occurs mainly through respiratory droplets, but they can also occur through contact with contaminated surfaces. Vaccine tests were carried out approved by the World Health Organization (WHO). Brazil stands out in second world position, with four approved vaccines: Pfizer-BioNTech, Oxford-AstraZeneca, CoronaVac (Sinovac), Janssen/Covishield.

A partir do primeiro caso de COVID-19 no Brasil, o país se tornou o terceiro no mundo em números de casos e de óbitos. Apesar das medidas implantadas pelo governo, o número de infectados e de óbitos por COVID-19 continua aumentando e o país enfrenta vários outros problemas que inclui aspectos sociais e políticos, dificultando as medidas de contenção da pandemia. O presente estudo visou abordar as características gerais do SARS-Cov-2, bem como apontar os principais aspectos socioepidemiológico no Brasil, e tratamento da COVID 19. Foi realizada uma revisão de literatura para busca de artigos em Bases de dados PubMed, Scielo e Google Scholar até 06 de outubro de 2020. Os pacientes com COVID-19 podem ser assintomáticos, porém entre os sintomáticos a gravidade da doença está relacionada à idade e a condições médicas pré-existentes. Os pulmões são os órgãos mais afetados pelo vírus e por isso as manifestações respiratórias como tosse, falta de ar, produção de escarro, dor de garganta e congestão nasal são os sintomas mais associados à COVID-19 A transmissão do SARS-COV-2 entre os humanos ocorre principalmente por meio de gotículas respiratórias, mas também podem ocorrer por meio do contato com superfícies contaminadas. Testes de vacinas foram realizados aprovados pela Organização Mundial da Saúde (OMS). O Brasil se destaca em segunda posição mundial, com cinco vacinas aprovadas, Pfizer-BioNTech, Oxford-AstraZeneca, /CoronaVac (Sinovac), Janssen/Covishield.