Virulence factors, antifungal susceptibility and molecular profile in Candida species isolated from the hands of health professionals before and after cleaning with 70% ethyl alcohol-based gel.

Fungal infections in neonatal intensive care units (NICU) are mainly related to Candida species, with high mortality rates. They are predominantly of endogenous origin, however, cross-infection transmitted by healthcare professionals' hands has occurred. The aim of this study was to identify Candida species isolated from the hands of healthcare professionals in a NICU before and after hygiene with 70% ethanol-based gel and evaluate virulence factors DNase, phospholipase, proteinase, hemolysin, biofilm biomass production, and metabolic activity. In vitro antifungal susceptibility testing and similarity by random amplified polymorphic DNA (RAPD) were also performed. C. parapsilosis complex was the most frequent species (57.1%); all isolates presented at least one virulence factor; three isolates (Candida parapsilosis complex) were resistant to amphotericin B, two (Candida famata [currently Debaryomyces hansenii] and Candida guilliermondii [currently Meyerozyma guilliermondii]) was resistant to micafungin, and six (Candida parapsilosis complex, Candida guilliermondii [=Meyerozyma guilliermondii], Candida viswanathi, Candida catenulata [currently Diutina catenulata] and Candida lusitaniae [currently Clavispora lusitaniae]) were resistant to fluconazole. Molecular analysis by RAPD revealed two clusters of identical strains that were in the hands of distinct professionals. Candida spp. were isolated even after hygiene with 70% ethanol-based gel, highlighting the importance of stricter basic measures for hospital infection control to prevent nosocomial transmission.

Antimicrobial, Antivirulence, and Antiparasitic Potential of Capsicum chinense Jacq. Extracts and Their Isolated Compound Capsaicin.

Bacterial, fungal, and parasitic infections increase morbimortality rates and hospital costs. This study aimed to assess the antimicrobial and antiparasitic activities of the crude extract from the seeds and peel of the pepper Capsicum chinense Jacq. and of the isolated compound capsaicin and to evaluate their ability to inhibit biofilm formation, eradicate biofilm, and reduce hemolysin production by Candida species. The crude ethanolic and hexane extracts were obtained by maceration at room temperature, and their chemical compositions were analyzed by liquid chromatography coupled to mass spectrometry (LC-MS). The antimicrobial activity of the samples was evaluated by determining the minimum inhibitory concentration. Inhibition of biofilm formation and biofilm eradication by the samples were evaluated based on biomass and cell viability. Reduction of Candida spp. hemolytic activity by the samples was determined on sheep blood agar plates. The antiparasitic action of the samples was evaluated by determining their ability to inhibit Toxoplasma gondii intracellular proliferation. LC-MS-ESI analyses helped to identify organic and phenolic acids, flavonoids, capsaicinoids, and fatty acids in the ethanolic extracts, as well as capsaicinoids and fatty acids in the hexane extracts. Antifungal action was more evident against C. glabrata and C. tropicalis. The samples inhibited biofilm formation and eradicated the biofilm formed by C. tropicalis more effectively. Sub-inhibitory concentrations of the samples significantly reduced the C. glabrata and C. tropicalis hemolytic activity. The samples only altered host cell viability when tested at higher concentrations; however, at non-toxic concentrations, they reduced T. gondii growth. In association with gold standard drugs used to treat toxoplasmosis, capsaicin improved their antiparasitic activity. These results are unprecedented and encouraging, indicating the Capsicum chinense Jacq. peel and seed extracts and capsaicin display antifungal and antiparasitic activities.

Inanimate Surfaces and Air Contamination with Multidrug Resistant Species of Staphylococcus in the Neonatal Intensive Care Unit Environment.

BACKGROUND: Contamination of the hospital environment with multi-resistant (MDR) Staphylococcus increases the risk of infection. The aim of this study is to identify the MDR species of Staphylococcus on inanimate surfaces, in air, and in clinical samples, and analyze the risk factors that correlate with the occurrence of infections in a Neonatal Intensive Care Unit. METHODS: Samples of inanimate surfaces and air were taken using a premoistened swab (0.9% sodium chloride) and spontaneous air sedimentation, respectively. The clinical isolates were recovered from infected neonates. The isolates (environmental and clinical) were identified by matrix-assisted laser desorption ionization-time of flight and the resistance profile was calculated using the disk diffusion agar technique. RESULTS: In total, 181 isolates were obtained, 93 from (surfaces), 18 from the air, and 70 clinical samples. S. epidermidis was the most frequent species (66.8%), and the failure rate in air cleaning was 100%. More than 60% of the isolates were MDR, and the majority of clinical isolates (60.4%) had a resistance profile identical to that of the environmental isolates. CONCLUSION: Staphylococcus spp. were found in most of the analyzed samples, with a high frequency of MDR isolates, demonstrating the importance of the hospital environment as a reservoir, and the need for infection control measures, and rational use of antimicrobials.

Antibacterial Activity of Isobavachalcone (IBC) Is Associated with Membrane Disruption.

Isobavachalcone (IBC) is a natural prenylated chalcone with a broad spectrum of pharmacological properties. In this work, we newly synthesized and investigated the antibacterial activity of IBC against Gram-positive, Gram-negative and mycobacterial species. IBC was active against Gram-positive bacteria, mainly against Methicillin-Susceptible Staphylococcus aureus (MSSA) and Methicillin-Resistant Staphylococcus aureus (MRSA), with minimum inhibitory concentration (MIC) values of 1.56 and 3.12 µg/mL, respectively. On the other hand, IBC was not able to act against Gram-negative species (MIC > 400 µg/mL). IBC displayed activity against mycobacterial species (MIC = 64 µg/mL), including Mycobacterium tuberculosis, Mycobacterium avium and Mycobacterium kansasii. IBC was able to inhibit more than 50% of MSSA and MRSA biofilm formation at 0.78 µg/mL. Its antibiofilm activity was similar to vancomycin, which was active at 0.74 µg/mL. In order to study the mechanism of the action by fluorescence microscopy, the propidium iodide (PI) and SYTO9 fluorophores indicated that IBC disrupted the membrane of Bacillus subtilis. Toxicity assays using human keratinocytes (HaCaT cell line) showed that IBC did not have the capacity to reduce the cell viability. These results suggested that IBC is a promising antibacterial agent with an elucidated mode of action and potential applications as an antibacterial drug and a medical device coating.

Impact of alcohol gel on hand bacteria in healthcare professionals / Impacto del gel de alcohol en las bacterias de las manos en los profesionales sanitarios / Impacto do álcool gel nas bactérias das mãos de profissionais de saúde

Background and objectives: Healthcare-Associated Infections are a problem reported by hospitals worldwide, increasing patient morbidity and mortality, prolonging hospitalization, and increasing health care costs. The hands of health professionals are still the main source of infections, making hand hygiene extremely important for spreading infection control. The objective of this study was to analyze the presence of bacteria on the hands of health professionals after hygiene with alcohol gel in a Neonatal Unit and describe the resistance of microorganisms to antimicrobials. Methods: Hand samples were collected using the modified glove-juice method on both occasions, before and after hand hygiene with alcohol gel. Bacteria were identified by MALDI-TOF and susceptibility tests according to Clinical and Laboratory Standards Institute document M100-E29. Results: A total of 214 samples were obtained, of which 104 (48.6%) showed bacterial growth before hand hygiene and 52 (24.3%) after hand hygiene with alcohol gel. There were 217 isolates from the cultures, of which coagulase-negative Staphylococcus was the most frequent with 41 (27.2%) and 24 (36.4%) positive cultures, respectively before and after hand hygiene. The second most frequent microorganism was Klebsiella pneumoniae with 32 (21.2%) and 16 (24.2%), respectively before and after hand hygiene. Multidrug resistance to antimicrobials was detected in 58.1% of gram-positive bacteria and in 34.3% of gram-negative bacteria. Conclusion: A decrease was observed, but not an elimination of the microbial load after hand hygiene with alcohol gel, demonstrating the need for improvements in hand hygiene.(AU)

Justificativa e objetivos: As Infecções Relacionadas à Assistência à Saúde são um problema relatado por hospitais em todo o mundo, aumentando a morbimortalidade dos pacientes, prolongando a hospitalização e aumentando os custos dos cuidados de saúde. As mãos dos profissionais de saúde ainda são a principal fonte de infecções, tornando a higienização das mãos extremamente importante para a disseminação do controle de infecções. O objetivo deste estudo foi analisar a presença de bactérias nas mãos de profissionais de saúde após higienização com álcool gel em uma Unidade Neonatal e descrever a resistência dos microrganismos aos antimicrobianos. Métodos: Amostras de mãos foram coletadas pelo método luva-suco modificado em ambas as ocasiões, antes e após a higienização das mãos com álcool gel. As bactérias foram identificadas por MALDI-TOF e testes de suscetibilidade de acordo com o documento M100-E29 do Clinical and Laboratory Standards Institute. Resultados: Obteve-se um total de 214 amostras, das quais 104 (48,6%) apresentaram crescimento bacteriano antes da higienização das mãos e 52 (24,3%) após a higienização das mãos com álcool gel. Foram 217 isolados das culturas, sendo Staphylococcus coagulase-negativo o mais frequente com 41 (27,2%) e 24 (36,4%) culturas positivas, respectivamente antes e após a higienização das mãos. O segundo microrganismo mais frequente foi Klebsiella pneumoniae com 32 (21,2%) e 16 (24,2%), respectivamente antes e após a higienização das mãos. A multirresistência aos antimicrobianos foi detectada em 58,1% das bactérias gram-positivas e em 34,3% das bactérias gram-negativas. Conclusão: Observou-se diminuição, mas não eliminação da carga microbiana após higienização das mãos com álcool gel, demonstrando a necessidade de melhorias na higienização das mãos.(AU)

Justificación y objetivos: Las Infecciones Asociadas a la Atención de la Salud son un problema reportado por los hospitales a nivel mundial, aumentando la morbimortalidad de los pacientes, prolongando la hospitalización y aumentando los costos de la atención médica. Las manos de los profesionales de la salud siguen siendo la principal fuente de infecciones, por lo que la higiene de manos es extremadamente importante para el control de infecciones. El objetivo de este estudio fue analizar la presencia de bacterias en las manos de los profesionales de la salud después de la higiene con alcohol en gel en una Unidad Neonatal y describir la resistencia de los microorganismos a los antimicrobianos. Métodos: Se recogieron muestras de manos mediante el método guante-jugo modificado en ambas ocasiones, antes y después de la higiene de manos con alcohol en gel. Las bacterias se identificaron mediante MALDI- -TOF y pruebas de susceptibilidad de acuerdo con el documento M100-E29 del Clinical and Laboratory Standards Institute. Resultados: Se obtuvieron un total de 214 muestras, de las cuales 104 (48,6%) presentaron crecimiento bacteriano antes de la higiene de manos y 52 (24,3%) después de la higiene de manos con alcohol en gel. Hubo 217 aislamientos de los cultivos, de los cuales el Staphylococcus coagulasa negativo fue el más frecuente con 41 (27,2%) y 24 (36,4%) cultivos positivos, respectivamente antes y después de la higiene de manos. El segundo microorganismo más frecuente fue Klebsiella pneumoniae con 32 (21,2%) y 16 (24,2%), respectivamente antes y después de la higiene de manos. Se detectó multirresistencia a los antimicrobianos en el 58,1% de las bacterias grampositivas y en el 34,3% de las bacterias gramnegativas. Conclusión: Se observó una disminución, pero no una eliminación de la carga microbiana después de la higiene de manos con alcohol en gel, lo que demuestra la necesidad de mejoras en la higiene de manos.(AU)

Copper(II) complexes based on thiosemicarbazone ligand: Preparation, crystal structure, Hirshfeld surface, energy framework, antiMycobacterium activity, in silico and molecular docking studies.

Considering the promising previous results on the remarkable activity exhibited by cobalt(III) and manganese(II) thiosemicarbazone compounds as antibacterial agents, the present study aimed to prepare and then evaluate the antibacterial activity of two different types of Cu(II) complexes based on a 2-acetylpyridine-N(4)-methyl-thiosemicarbazone ligand (Hatc-Me), a monomer complex [CuCl(atc-Me)] and a novel dinuclear complex [{Cu(µ-atc-Me)}2µ-SO4]. The compounds were characterized by infrared spectra, ultraviolet visible and CHN elemental analysis. In addition, the crystalline structures of the complexes were determined by single-crystal X-ray diffraction. In both cases, the Schiff base ligand coordinated in a tridentate mode via the pyridine nitrogen, imine nitrogen and sulfur atoms. The two Cu(II) atoms in the dimer are five coordinate, consisting of three NNS-donor atoms from the thiosemicarbazone ligand connected by a sulfate bridge. The Hirshfeld surface and energy framework of the complexes were additionally analyzed to verify the intermolecular interactions. The biological activity of the Cu(II) salts, the free ligand and its Cu(II) complexes was evaluated against six strains of mycobacteria including Mycobacterium tuberculosis. The complexes showed promising results as antibacterial agents for M. avium and M. tuberculosis, which ranged from 6.12 to 12.73 µM. Furthermore, molecular docking analysis was performed and the binding energy of the docked compound [{Cu(µ-atc-Me)}2µ-SO4] with M. tuberculosis and M. avium strains were extremely favorable (-11.11 and - 14.03 kcal/mol, respectively). The in silico results show that the complexes are potential candidates for the development of new antimycobacterial drugs.

Risk factors associated with mortality among patients who had candidemia in a university hospital.

INTRODUCTION: Bloodstream infection due to Candida spp. is a primary cause of morbidity and mortality in tertiary hospitals. METHODS: In this retrospective study, we included patients with a positive blood culture for Candida spp. after 48 h of hospitalization. RESULTS: A total of 335 patients who had candidemia were included in this study. Risk factors associated with mortality were hospitalization in internal medicine units and surgical clinics, age >60 years, mechanical ventilation, orotracheal intubation, hemodialysis, corticosteroids use, and C. parapsilosis infection. CONCLUSIONS: This study highlights the importance of health care related to invasive procedures and actions to improve patient immunity.

Candidemia by Candida parapsilosis in a neonatal intensive care unit: human and environmental reservoirs, virulence factors, and antifungal susceptibility.

The Candida parapsilosis complex has emerged as one of the main causes of candidemia worldwide. This study aims to evaluate possible C. parapsilosis sensu stricto reservoirs in a NICU, the expression of virulence factors, and antifungal susceptibility, and to analyze their genetic and phenotypic similarity. The study included 17 isolates of C. parapsilosis: seven environmental, one from a newborn's mother, and nine samples from six newborns. We used molecular and phenotypic tests to characterize the isolates and to trace possible routes of infection. The genetic similarity was determined by random amplified polymorphic DNA. The hemolytic and DNAse activity was determined using sheep's blood and DNAse agar, biofilm production by XTT method, and the susceptibility to antifungals through microdilution methodology. Two environmental strains isolated in the same month had high similarity. The 17 isolates expressed at least one of the three virulence factors studied, and one environmental isolate was resistant to fluconazole. This study shows that environmental contamination can be an important reservoir of potentially pathogenic microorganisms, since isolates of C. parapsilosis sensu stricto collected from the hospital environment were able to express virulence factors. Therefore, we emphasized the importance of determining the transmission routes in NICU in order to detect pathogen sources and reservoirs, as well as to establish prevention measures, such as adequate disinfection of the environment.

Risk factors associated with mortality among patients who had candidemia in a university hospital

Abstract INTRODUCTION: Bloodstream infection due to Candida spp. is a primary cause of morbidity and mortality in tertiary hospitals. METHODS: In this retrospective study, we included patients with a positive blood culture for Candida spp. after 48 h of hospitalization. RESULTS A total of 335 patients who had candidemia were included in this study. Risk factors associated with mortality were hospitalization in internal medicine units and surgical clinics, age >60 years, mechanical ventilation, orotracheal intubation, hemodialysis, corticosteroids use, and C. parapsilosis infection. CONCLUSIONS: This study highlights the importance of health care related to invasive procedures and actions to improve patient immunity.

Isolation of Cryptococcus species from the external environments of hospital and academic areas.

INTRODUCTION: Fungi of the genus Cryptococcus are cosmopolitan and may be agents of opportunistic mycoses in immunocompromised and sometimes immunocompetent individuals. Cryptococcus species are frequently isolated from trees and bird excreta in the environment and infection occurs by inhalation of propagules dispersed in the air. The aim was to investigate Cryptococcus species in bird excreta and tree hollows located in a university hospital area and in an academic area of a university campus. METHODOLOGY: A total of 40 samples of bird excreta and 41 samples of tree hollows were collected. The identification of the isolates was done by classical methodology and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. RESULTS: Twenty (62.5%) isolates of Cryptococcus were found in bird excreta and 12 (37.5%) in tree hollows. C. laurentii (currently Papiliotrema laurentii) was the most frequent species in both samples, being found in 5 samples of excreta and in 8 tree hollows. The diversity of species found in excreta (C. laurentii, C. albidus [currently Naganishia albida], C. liquefaciens [currently N. liquefaciens], C. friedmanii [currently N. friedmannii] and others) was higher than in tree hollows (C. laurentii, C. flavescens [currently Papiliotrema flavescens], and other yeasts). CONCLUSION: Many Cryptococcus species were isolated from excreta and tree hollows, and this fact is important for understanding the environmental epidemiology of those emerging pathogens for public health, as a way to implement surveillance actions and control of cryptococcosis.

Characterization of Candida species isolated from the hands of the healthcare workers in the neonatal intensive care unit.

Colonization of health professional hands by potentially pathogenic microorganisms capable to expressing virulence factors, such as Candida spp., is worrisome because of the high contact between patients and professionals. The study aims to evaluate, in vitro, the following virulence factors: hemolytic activity, DNAse expression, biofilm formation, and susceptibility for antifungal agents of Candida species isolated from health professionals hands of a neonatal intensive care unit (NICU). The study includes 50 isolates of Candida spp.: 19 C. parapsilosis sensu stricto, three C. metapsilosis, one C. orthopsilosis, seven C. albicans, six C. famata, five C. lusitaniae, three C. krusei, two C. kefyr, two C. tropicalis, one C. glabrata, and one C. guilliermondii. The hemolytic activity and DNAse were investigated using blood agar and DNAse agar, respectively. Biofilm production was evaluated through XTT sodium salt reduction ability, and the susceptibility of the isolates to antifungals through the microdilution methodology. Forty-nine isolates presented at least one of the three virulence factors investigated. C. albicans showed more intense hemolytic activity. DNAse production was statistically significant between the C. parapsilosis complex and C. albicans, as well as between the C. parapsilosis complex and C. krusei, even as between C. famata and C. albicans, and between C. famata and C. krusei. Forty-three isolates produced biofilm. Seventy-eight percent of the isolates were sensitive to the three antifungals tested. This study demonstrated that Candida isolated from healthcare professionals' hands has virulence.