RESUMO
INTRODUCTION: Serratia marcescens is an opportunistic pathogen found ubiquitously in the environment and associated with a wide range of nosocomial infections. This multidrug-resistant bacterium has been a cause of concern for hospitals and healthcare facilities due to its ability to spread rapidly and cause outbreaks. Next generation sequencing genotyping of bacterial isolates has proven to be a valuable tool for tracking the spread and transmission of nosocomial infections. This has allowed for the identification of outbreaks and transmission chains, as well as determining whether cases are due to endogenous or exogenous sources. Evidence of nosocomial transmission has been gathered through genotyping methods. The aim of this study was to investigate the genetic diversity of carbapenemase-producing S. marcescens in an outbreak at a public hospital in Cuiaba, MT, Brazil. METHODOLOGY: Ten isolates of S. marcenses were sequenced and antibiotic resistance profiles analyzed over 12 days. RESULTS: The isolates were clonal and multidrug resistant. Gentamycin and tigecycline had sensitivity in 90% and 80% isolates, respectively. Genomic analysis identified several genes that encode ß-lactamases, aminoglycoside-modifying enzymes, efflux pumps, and other virulence factors. CONCLUSIONS: Systematic surveillance is crucial in monitoring the evolution of S. marcescens genotypes, as it can lead to early detection and prevention of outbreaks.
Assuntos
Antibacterianos , Infecção Hospitalar , Surtos de Doenças , Farmacorresistência Bacteriana Múltipla , Unidades de Terapia Intensiva , Infecções por Serratia , Serratia marcescens , Sequenciamento Completo do Genoma , Serratia marcescens/genética , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/isolamento & purificação , Humanos , Brasil/epidemiologia , Farmacorresistência Bacteriana Múltipla/genética , Infecções por Serratia/microbiologia , Infecções por Serratia/epidemiologia , Infecção Hospitalar/microbiologia , Infecção Hospitalar/epidemiologia , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Genótipo , Genoma Bacteriano , beta-Lactamases/genética , Variação GenéticaRESUMO
The bacterial composition of and the circulation of antimicrobial resistance genes (ARGs) in waste from Brazilian swine farms are still poorly understood. Considering that antimicrobial resistance (AMR) is one of the main threats to human, animal, and environmental health, the need to accurately assess the load of ARGs released into the environment is urgent. Therefore, this study aimed to characterize the microbiota in a swine farm in southern Brazil and the resistome in swine farm wastewater treated in a series of waste stabilization ponds (WSPs). Samples were collected from farm facilities and the surrounding environment, representing all levels of swine manure within the treatment system. Total metagenomic sequencing was performed on samples from WSPs, and 16S-rDNA sequencing was performed on all the collected samples. The results showed increased bacterial diversity in WSPs, characterized by the presence of Caldatribacteriota, Cloacimonadota, Desulfobacterota, Spirochaetota, Synergistota, and Verrucomicrobiota. Furthermore, resistance genes to tetracyclines, lincosamides, macrolides, rifamycin, phenicol, and genes conferring multidrug resistance were detected in WSPs samples. Interestingly, the most abundant ARG was linG, which confers resistance to the lincosamides. Notably, genes conferring macrolide (mphG and mefC) and rifamycin (rpoB_RIF) resistance appeared in greater numbers in the late WSPs. These drugs are among the high-priority antibiotic classes for human health. Moreover, certain mobile genetic elements (MGEs) were identified in the samples, notably tnpA, which was found in high abundance. These elements are of particular concern due to their potential to facilitate the dissemination of ARGs among bacteria. In summary, the results indicate that, in the studied farm, the swine manure treatment system could not eliminate ARGs and MGEs. Our results validate concerns about Brazil's swine production system. The misuse and overuse of antimicrobials during animal production must be avoided to mitigate AMR.
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Antibacterianos , Bactérias , Farmacorresistência Bacteriana , Fazendas , Animais , Suínos , Brasil , Bactérias/genética , Bactérias/efeitos dos fármacos , Farmacorresistência Bacteriana/genética , Antibacterianos/farmacologia , Genes Bacterianos , Águas Residuárias/microbiologia , Esterco/microbiologia , Microbiota/efeitos dos fármacos , Microbiota/genéticaRESUMO
In herds of dairy goats, mastitis represents a major health and economic problem due to the multiresistance of some microorganisms. In this context, the study aimed to determine the potential of antimicrobial action and antibiofilm of the crude ethanolic extract (CEE) of Hymenaea martiana (jatobá) leaves, as well its fractions, on Staphylococcus sp isolated from bacterial cultures of goat milk. In vitro assays were performed to determine the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC), as well as tests of the effect of CEE on biofilm formation and quantification and the consolidated biofilm. The experimental infection was performed in two groups, each consisting of five goat. Experimental Group 1 (G1) consisted of five females treated with an intramammary ointment based on the CEE, at a concentration of 5%. Experimental Group 2 (G2) consisted of five females treated with a commercial intramammary ointment based on gentamicin, once a day, for six consecutive days. The diagnosis of mastitis was performed using a bacterial culture. The dichloromethane fraction of CEE was the one with the lowest concentrations of MBC, ranging from 195.3 to 781 µg / ml. Concerning to the biofilm, interference of the tested extract was observed for two isolates. In the present study, the ointment prepared from H. martiana extract (jatobá) was able to reduce bacterial infection in mammary glands experimentally infected with S. aureus. Antibacterial activity may be related to the classes of secondary metabolites found.
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Antibacterianos , Biofilmes , Doenças das Cabras , Cabras , Mastite , Testes de Sensibilidade Microbiana , Extratos Vegetais , Infecções Estafilocócicas , Staphylococcus aureus , Animais , Feminino , Doenças das Cabras/tratamento farmacológico , Doenças das Cabras/microbiologia , Infecções Estafilocócicas/veterinária , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/fisiologia , Mastite/veterinária , Mastite/tratamento farmacológico , Mastite/microbiologia , Testes de Sensibilidade Microbiana/veterinária , Antibacterianos/farmacologia , Antibacterianos/administração & dosagem , Extratos Vegetais/farmacologia , Extratos Vegetais/administração & dosagem , Biofilmes/efeitos dos fármacos , Leite/microbiologia , Folhas de Planta/químicaRESUMO
In this study, the main agents associated with endometritis in cows in the state of Santa Catarina, Brazil, were identified and the resistance profile and virulence mechanisms of the bacterial isolates were evaluated. Isolates of Escherichia coli and Trueperella pyogenes were tested for their biofilm forming ability and the antimicrobial action of bromhexine hydrochloride in combination with other antimicrobials. A total of 37 uterine lavage samples were collected from cows with endometritis. Of the 55 bacteria isolated, 25.4% were identified as T. pyogenes and 16.3% as E. coli. The bacterial isolates showed greater resistance to sulfamethoxazole + trimethoprim (58.2%) and tetracycline (56.3%). Among the species, E. coli showed the highest resistance rates, with 100% of isolates showing resistance to amoxicillin, streptomycin, and gentamicin. The results of the minimum inhibitory concentration for the T. pyogenes isolates showed that 91.6% of the isolates were resistant to enrofloxacin and tetracycline, and 75% were resistant to ceftiofur and sulfamethoxazole + trimethoprim. All E. coli and T. pyogenes isolates showed biofilm forming ability. The plo, fimA, and nanH genes were identified in 100% of T. pyogenes isolates. In parallel, 100% of E. coli isolates had the fimH gene, and 11.1% had the csgD gene. Bromhexine hydrochloride showed antimicrobial activity against 100% of E. coli isolates and 66.6% of T. pyogenes isolates. Furthermore, when associated with antimicrobials, bromhexine hydrochloride has a synergistic and additive effect, proving to be an option in the treatment of endometritis in cows and an alternative for reducing the use of antimicrobials.
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Actinomycetaceae , Infecções por Actinomycetales , Antibacterianos , Doenças dos Bovinos , Endometrite , Infecções por Escherichia coli , Escherichia coli , Testes de Sensibilidade Microbiana , Bovinos , Animais , Endometrite/microbiologia , Endometrite/veterinária , Endometrite/tratamento farmacológico , Feminino , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Antibacterianos/farmacologia , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/tratamento farmacológico , Actinomycetaceae/efeitos dos fármacos , Actinomycetaceae/genética , Actinomycetaceae/isolamento & purificação , Actinomycetaceae/classificação , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/veterinária , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Actinomycetales/microbiologia , Infecções por Actinomycetales/veterinária , Infecções por Actinomycetales/tratamento farmacológico , Biofilmes/efeitos dos fármacos , Brasil , Farmacorresistência BacterianaRESUMO
Cardiac maturation represents the last phase of heart development and is characterized by morphofunctional alterations that optimize the heart for efficient pumping. Its understanding provides important insights into cardiac regeneration therapies. Recent evidence implies that adrenergic signals are involved in the regulation of cardiac maturation, but the mechanistic underpinnings involved in this process are poorly understood. Herein, we explored the role of ß-adrenergic receptor (ß-AR) activation in determining structural and functional components of cardiomyocyte maturation. Temporal characterization of tyrosine hydroxylase and norepinephrine levels in the mouse heart revealed that sympathetic innervation develops during the first 3 wk of life, concurrent with the rise in ß-AR expression. To assess the impact of adrenergic inhibition on maturation, we treated mice with propranolol, isolated cardiomyocytes, and evaluated morphofunctional parameters. Propranolol treatment reduced heart weight, cardiomyocyte size, and cellular shortening, while it increased the pool of mononucleated myocytes, resulting in impaired maturation. No changes in t-tubules were observed in cells from propranolol mice. To establish a causal link between ß-AR signaling and cardiomyocyte maturation, mice were subjected to sympathectomy, followed or not by restoration with isoproterenol treatment. Cardiomyocytes from sympathectomyzed mice recapitulated the salient immaturity features of propranolol-treated mice, with the additional loss of t-tubules. Isoproterenol rescued the maturation deficits induced by sympathectomy, except for the t-tubule alterations. Our study identifies the ß-AR stimuli as a maturation promoting signal and implies that this pathway can be modulated to improve cardiac regeneration therapies.NEW & NOTEWORTHY Maturation involves a series of morphofunctional alterations vital to heart development. Its regulatory mechanisms are only now being unveiled. Evidence implies that adrenergic signaling regulates cardiac maturation, but the mechanisms are poorly understood. To address this point, we blocked ß-ARs or performed sympathectomy followed by rescue experiments with isoproterenol in neonatal mice. Our study identifies the ß-AR stimuli as a maturation signal for cardiomyocytes and highlights the importance of this pathway in cardiac regeneration therapies.
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Miócitos Cardíacos , Propranolol , Transdução de Sinais , Animais , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/efeitos dos fármacos , Camundongos , Propranolol/farmacologia , Receptores Adrenérgicos beta/metabolismo , Camundongos Endogâmicos C57BL , Isoproterenol/farmacologia , Masculino , Coração/efeitos dos fármacos , Células Cultivadas , Agonistas Adrenérgicos beta/farmacologia , Norepinefrina/metabolismo , Norepinefrina/farmacologia , Antagonistas Adrenérgicos beta/farmacologiaRESUMO
OBJECTIVE: To validate the Cancer of the Bladder Risk Assessment (COBRA) score in patients with urothelial variants. METHODS: Epidemiological, clinical, radiological, and anatomopathological data were collected from patients with urothelial carcinoma who underwent radical cystectomy at the Institute of Cancer of São Paulo between May 2008 and December 2022. Patients with the presence of at least 10% of any urothelial variants in the radical cystectomy specimens' anatomopathological exam were included in the study. The COBRA score and derivatives were applied and correlated with oncological outcomes. RESULTS: A total of 680 patients [482 men (70.9%) and 198 women (29.1%)]; 66 years (IQR 59-73) underwent radical cystectomy for bladder tumor, and of these patients, a total of 167 patients presented any type of urothelial variant. The median follow-up time was 28.77 months (IQR 12-85). The three most prevalent UV were squamous differentiation (50.8%), glandular differentiation (31.3%), and micropapillary differentiation (11.3%). The subtypes with the worst prognosis were sarcomatoid with a median survival of 8 months (HR 1.161; 95% CI 0.555-2.432) and plasmacytoid with 14 months (HR 1.466; 95% CI 0.528-4.070). The COBRA score for patients with micropapillary variants demonstrated good predictive accuracy for OS (log-rank P = 0.009; 95% IC 6.78-29.21) and CSS (log-rank P = 0.002; 95% IC 13.06-26.93). CONCLUSIONS: In our study, the COBRA score proved an effective risk stratification tool for urothelial histological variants, especially for the micropapillary urothelial variant. It may be helpful in the prognosis evaluation of UV patients after radical cystectomy.
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Carcinoma de Células de Transição , Neoplasias da Bexiga Urinária , Masculino , Humanos , Feminino , Neoplasias da Bexiga Urinária/patologia , Carcinoma de Células de Transição/patologia , Cistectomia , Estudos Retrospectivos , Brasil , Medição de RiscoRESUMO
AIMS: The purpose was to evaluate the antimicrobial activity of highly soluble polypyrrole (Hs-PPy), alone or combined with oxacillin, as well as its antibiofilm potential against methicillin-resistant Staphylococcus aureus strains. Furthermore, the in silico inhibitory mechanism in efflux pumps was also investigated. METHODS AND RESULTS: Ten clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and two reference strains were used. Antimicrobial activity was determined by broth microdilution, and the combination effect with oxacillin was evaluated by the checkerboard assay. The biofilm formation capacity of MRSA and the interference of Hs-PPy were evaluated. The inhibitory action of Hs-PPy on the efflux pump was evaluated in silico through molecular docking. Hs-PPy showed activity against the isolates, with inhibitory action between 62.5 and 125 µg ml-1 and bactericidal action at 62.5 µg ml-1, as well as synergism in association with oxacillin. The isolates ranged from moderate to strong biofilm producers, and Hs-PPy interfered with the formation of this structure, but not with mature biofilm. There was no in silico interaction with the efflux protein EmrD, the closest homolog to NorA. CONCLUSIONS: Hs-PPy interferes with biofilm formation by MRSA, has synergistic potential, and is an efflux pump inhibitor.
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Anti-Infecciosos , Staphylococcus aureus Resistente à Meticilina , Antibacterianos/farmacologia , Polímeros/farmacologia , Pirróis/farmacologia , Simulação de Acoplamento Molecular , Oxacilina/farmacologia , Anti-Infecciosos/farmacologia , Biofilmes , Testes de Sensibilidade MicrobianaRESUMO
Escherichia coli (E. coli) are widely related to pyometra and cystitis in dogs, and these infections can occur simultaneously. The goal of this study was to determine genetic and pathogenic insights of 14 E. coli isolated simultaneously from pyometra content and bladder urine of seven bitches. To achieve this, in silico and in vitro comparative analyses were conducted. Whole-genome comparisons demonstrated that E. coli isolated from pyometra and urine of the same animal were predominantly genetic extraintestinal E. coli clones belonging to the same Sequence Type and phylogroup. The E. coli clones identified in this study included ST372, ST457, ST12, ST127, ST646, and ST961. Five isolates (35.7%) belonged to the ST12 complex. Except for two E. coli, all other isolates belonged to the B2 Clermont phylogroup. Interestingly, some genomes of E. coli from urine carried more virulence genes than those E. coli from pyometra. Both pyometra and urine E. coli isolates demonstrated a strong affinity for adhering to HeLa and T24 cells, with a low affinity for invading them. However, certain isolates from urine exhibited a greater tendency to adhere to T24 cells in qualitative and quantitative assays compared to isolates from pyometra. In conclusion, this study revealed the high genomic similarity between pyometra and urine E. coli isolates, as well as the virulent capacity of both to colonize endometrial and urothelial cells. The findings of this study underscore the importance of concurrently managing both infections clinically and could potentially contribute to future resources for the prevention of cystitis and pyometra.
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Doenças do Cão , Infecções por Escherichia coli , Escherichia coli , Piometra , Animais , Cães , Piometra/veterinária , Piometra/microbiologia , Piometra/urina , Feminino , Doenças do Cão/microbiologia , Doenças do Cão/urina , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Escherichia coli/patogenicidade , Infecções por Escherichia coli/veterinária , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/urina , Genoma Bacteriano , FilogeniaRESUMO
Mastitis caused by Staphylococcus aureus is a worldwide problem in dairy farms, in part because of the pathogenicity of the bacteria, biofilm formation, and mechanisms of antimicrobial resistance that make the disease difficult to diagnose and treat, which is typically done with the use of beta-lactam antibiotics. The aim of the present study was to determine the virulence and resistance factors of S. aureus isolates from subclinical mastitis, blaZ + /mecA - /mecC - , resistant and sensitive to oxacillin. All isolates were classified as CC97 by MLST analysis, a clonal complex well adapted to the mammary gland and although STAU23 and STAU73 were resistant to oxacillin while STAU32 and STAU78 were sensitive, the genomic analysis identified only the blaZ operon corresponding to resistance to beta-lactams. However, the presence of the sdrC gene was revealed exclusively in resistant isolates, an important adhesin in the colonization process that potentiates pathogenicity in S. aureus. In addition, resistance islands (REIs) were identified in these isolates, suggesting more conserved REIs. In the analysis of SNPs throughout the genome, mutations were found in the trmB and smpB genes of the resistant isolates and in the murD and rimM genes of the sensitive isolates. This study highlights the potential benefit of genome-wide characterization tools to identify molecular mechanisms of S. aureus in bovine mastitis.
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Mastite Bovina , Infecções Estafilocócicas , Animais , Bovinos , Feminino , Humanos , Staphylococcus aureus , Antibacterianos/farmacologia , Virulência/genética , Tipagem de Sequências Multilocus , Testes de Sensibilidade Microbiana , Mastite Bovina/microbiologia , Infecções Estafilocócicas/veterinária , Infecções Estafilocócicas/microbiologia , OxacilinaRESUMO
The aim of the study was to track the spread of antimicrobial resistance among the different sectors of One Health through the detection of Multidrug-Efflux-System in multidrug-resistant Staphylococcus aureus isolates. Multidrug-resistant (MDR) and methicillin-resistant (MRSA) S. aureus isolates were selected: 25 of human, one of animal and eight of food origin. The efflux system genes norA, norB, norC, LmrS, tet38 and msrA were screened by PCR. The activity of the efflux systems was determined by the minimum inhibitory concentration (MIC) of tetracycline and ciprofloxacin in the presence and absence of CCCP and in the quantification of ethidium bromide efflux. Furthermore, biofilm formation was determined in the presence and absence of the CCCP. The molecular epidemiology of the isolates was traced with the aid of PFGE. The gene norC was the most prevalent, detected in all isolates and msrA was the least prevalent, detected in only two isolates from humans. There was no difference in the MICs of tetracycline and ciprofloxacin in the presence of CCCP, but 55.9% of isolates showed ethidium bromide efflux. The presence of CCCP decreased the biofilm formation. Regarding the molecular epidemiology, in three clusters was a mixture of the isolates from different origins. Therefore, S. aureus MDR with active multidrug efflux systems are circulating between One Health domains and it is necessary to consider strategies to decrease this circulation in order to prevent the dissemination of resistance mediated by MES.