Rapid diagnostic of multidrug-resistant sepsis pathogens directly from blood culture bottles using MALDI-TOF and the EUCAST RAST.

In this study, rapid diagnostic of multidrug-resistant (MDR) sepsis pathogens, directly from positive blood culture (BC) bottles, was evaluated by combining MALDI-TOF and the EUCAST Rapid Antimicrobial Susceptibility Testing (RAST). Carbapenemase production in Escherichia coli and Klebsiella pneumoniae isolates was also evaluated by RAST. From 171 positive BC bottles analyzed, 79 (46 %) MDR species, including E. coli (4/34, 12 %), K. pneumoniae (33/48, 69 %), Pseudomonas aeruginosa (12/12, 100 %), Acinetobacter baumannii (15/15, 100 %), and Staphylococcus aureus (14/37, 38 %) displaying resistance to beta-lactams, fluoroquinolones, aminoglycosides, and/or trimethoprim/sulphamethoxazole, were identified. In this regard, turnaround time of direct MALDI-TOF identification and RAST was < 7 h, which was significantly (p< 0.05) lower than our routine method. Carbapenemase detection by RAST displayed 100% sensitivity and 88.7 % specificity at 8 h. This protocol could offer advantages for the treatment and clinical outcomes of septic patients, improving the rapid diagnostic of sepsis by MDR pathogens.

Emergence of KPC-113 and KPC-114 variants in ceftazidime-avibactam-resistant Klebsiella pneumoniae belonging to high-risk clones ST11 and ST16 in South America.

Two novel variants of Klebsiella pneumoniae carbapenemase (KPC) associated with resistance to ceftazidime-avibactam (CZA) and designated as KPC-113 and KPC-114 by NCBI were identified in 2020, in clinical isolates of Klebsiella pneumoniae in Brazil. While K. pneumoniae of ST16 harbored the blaKPC-113 variant on an IncFII-IncFIB plasmid, K. pneumoniae of ST11 carried the blaKPC-114 variant on an IncN plasmid. Both isolates displayed resistance to broad-spectrum cephalosporins, ß-lactam inhibitors, and ertapenem and doripenem, whereas K. pneumoniae producing KPC-114 showed susceptibility to imipenem and meropenem. Whole-genome sequencing and in silico analysis revealed that KPC-113 presented a Gly insertion between Ambler positions 264 and 265 (R264_A265insG), whereas KPC-114 displayed two amino acid insertions (Ser-Ser) between Ambler positions 181 and 182 (S181_P182insSS) in KPC-2, responsible for CZA resistance profiles. Our results confirm the emergence of novel KPC variants associated with resistance to CZA in international clones of K. pneumoniae circulating in South America. IMPORTANCE KPC-2 carbapenemases are endemic in Latin America. In this regard, in 2018, ceftazidime-avibactam (CZA) was authorized for clinical use in Brazil due to its significant activity against KPC-2 producers. In recent years, reports of resistance to CZA have increased in this country, limiting its clinical application. In this study, we report the emergence of two novel KPC-2 variants, named KPC-113 and KPC-114, associated with CZA resistance in Klebsiella pneumoniae strains belonging to high-risk clones ST11 and ST16. Our finding suggests that novel mutations in KPC-2 are increasing in South America, which is a critical issue deserving active surveillance.

Assessment of phytotoxic potential and pathogenic bacteria removal from secondary effluents during ozonation and UV/H2O2.

Wastewater reuse is an important strategy for water resource management. For this reason, the disinfection process must be appropriated, eliminating pathogenic microorganisms. Ozonation (O3) and UV/H2O2 treatments can be used for effluent disinfection, but few studies just address the Escherichia coli quantification. In this study, secondary effluents from two wastewater treatment plants with different characteristics were exposed to O3 (5 and 10 mg L-1) or UV/H2O2 (H2O2: 90 mg L-1) treatments and evaluated by BD Phoenix ™ 100 (Becton Dickinson, USA) and MALDI-TOF for the characterization of the indigenous microorganisms in the effluents, before and after treatments. Additionally, all the samples were tested for phytotoxicity by Lactuca sativa bioassay. The results showed that the highest ozone dose and the UV/H2O2 treatment were effective in removing E. coli. UV/H2O2 was more efficient as it eliminated most of the microorganisms. Acinetobacter sp., Aeromonas and Pseudomonas were still found after O3 treatment. Bacillus sp. was found after O3 and UV/H2O2 treatments. The results with L. sativa showed inhibition of root growth for all dry period (low rainfall) samples of one of the WWTP, due to the high concentration of the phytotoxicity compounds. For environmental and human health safety, treated effluents should be evaluated for their toxic and pathogenic potential before being released into the environment. Pathogens evaluation on treated effluents should cover a wider range of pathogenic microorganisms than those routinely required by legislation.

Genomic analysis of a Kpi (pilus system)-positive and CTX-M-15-producing Klebsiella pneumoniae belonging to the high-risk clone ST15 isolated from an impacted river in Brazil.

Convergence of resistance and virulence in Klebsiella pneumoniae is a critical public health issue worldwide. A multidrug-resistant CTX-M-15-producing K. pneumoniae (TIES-4900 strain) was isolated from a highly impacted urban river, in Brazil. The genome was sequenced by MiSeq Illumina platform and de novo assembled using Unicycler. In silico prediction was accomplished by bioinformatics tools. The size of the genome is 5.4 Mb with 5145 protein-coding genes. TIES-4900 strain belonged to the sequence type ST15, yersiniabactin sequence type YbST10, ICEKp4, KL24 (wzi-24) and O1v1 locus. Phylogenomics confirmed genomic relatedness with ST15 clones from human and animal hosts. Convergence of broad resistome (antibiotics, heavy-metals and biocides) and virulome, including the Kpi pilus system involved in host-pathogen interaction and persistence of ST15 clone to hospital environments, were predicted. Virulent behavior was confirmed in the Galleria mellonella infection model. This study may give genomic insights on the spread of critical-priority WHO pathogens beyond hospital settings.

FONA-7, a Novel Extended-Spectrum ß-Lactamase Variant of the FONA Family Identified in Serratia fonticola.

Serratia fonticola is a human pathogen widely found in the environment, with birds being reported as possible natural hosts. During an epidemiological and genomic surveillance study conducted to monitor the occurrence of extended-spectrum ß-lactamase (ESBL)-producing Enterobacterales in South American wild birds, we identified an ESBL-positive S. fonticola in a fecal sample collected from a Hudsonian Whimbrel, during its non-breeding range on the Pacific Coast of Chile. Whole genome sequencing analysis and "in silico" modeling revealed a novel variant of the class A ESBLs FONA family, designated FONA-7, which shows 96.28% amino acid identity with FONA-6; with amino acid substitutions occurring in the signal peptide sequence (Thr22→Ser), and in the mature protein (Ser39→Asn and Thr227→Ile). This finding denotes that migratory birds can be potential vectors for the transboundary spread of ESBL-producing bacteria, creating a further theoretical risk for the origin of novel plasmid-encoded ß-lactamases.

Correlation between thermal, optical and morphological properties of heterogeneous blends of poly(3-hexylthiophene) and thermoplastic polyurethane.

A correlation between thermal, optical and morphological properties of self-sustained films formed from blends of poly(3-hexylthiophene) (P3HT) and thermoplastic polyurethane (TPU), with 1, 10 and 20 wt% of P3HT in TPU, is established. Images of scanning electron microscopy (SEM) show the formation of domains of P3HT into the TPU matrix, characterizing the blend material as heterogeneous. The heat capacity (C(p)) dependence on P3HT contents was investigated in a large temperature interval. In the region of the TPU glass transition, the difference between the experimental and predicted ΔC(p) values is more pronounced for the 1 wt% case, which strongly suggests that in this case there is a higher influence of the P3HT chains on the TPU matrix. The SEM images for the 1 wt% blended film present the formation of the smallest P3HT domains in the TPU matrix. The relatively high reduction of the PL intensity of the pure electronic transition peak in the 1 wt% blended film, in comparison to the other blended films and also to a pure P3HT film, favours the assumption that the smallest P3HT domains are at the origin of a more structural disordered character. This fact is in agreement with the results obtained by Raman spectroscopy and also by photoluminescence resolved by polarization in stretched self-sustained films, showing an ample correlation between morphological, thermal and optical properties of these blended materials. In addition, the thermoplastic properties of the polyurethane configure very good conditions for tensile drawing of P3HT and other conjugated polymer molecules.

A prevençäo e o diagnóstico de tétano: Uma responsabilidade do dentista? / Tetanus Prevention and diagnosis

O tétano é uma doença que mata cerca de mil pessoas por ano no Brasil. A maioria dos casos de tétano tem origem acidental por algum objeto que produza uma lesäo por meio da qual ocorrerá a penetraçäo do Clostridium tetani. Na Odontologia, a alta prevalência de traumas dento-alveolares associada às laceraçöes dos tecidos moles adjacentes pode propiciar uma porta de entrada para os microrganismos. O primeiro sintoma que o paciente apresenta é o trismo. Devido a este quadro clínico o cirurgiäo dentista tende a ser o primeiro profissional a ser procurado no momento inicial da doença. Desta forma, visando um maior esclarecimento sobre o assunto, este artigo tem como objetivo abordar, de forma sucinta, alguns aspectos básicos sobre a vacinaçäo e o diagnóstico do tétano