Use of Ozonized Water in the Prevention of Surgical Site Infection in Children Undergoing Cardiovascular Surgery.

INTRODUCTION: Since the reduction of healthcare-associated infections has been a focus for quality patient care, this study aimed to evaluate the surgical site infection rate of children who underwent cardiovascular surgery after implementation of ozonized water system for hand and body hygiene allied to previously implemented preventive measures. METHODS: Two uniformly comparable groups of pediatric patients underwent cardiovascular surgery. Group A (187) patients were operated prior to installation of ozonized water system (March 1 to August 31, 2019), and group B (214) patients were operated after installation of ozonized water system (October 1, 2019, to March 31, 2020). Ozonized water was used for professional hand hygiene and patient body hygiene. RESULTS: There was statistical significance for surgical site infection reduction in group B (P=0.0289), with a relative risk of 0.560 (95% confidence interval = 0.298 to 0.920), inferring the risk of being diagnosed with surgical site infections in group B was 44% less than in group A. There was no statistical significance regarding mechanical ventilation time (P=0.1998) or mortality (P=0.4457). CONCLUSION: Ozonized water for professional hand hygiene and patient body hygiene was an adjuvant combined with traditional preventive methods to reduce the risk of surgical site infection, although no impact on hospital stay or mortality was observed.

Combinatorial effect of fluconazole, itraconazole, and terbinafine with different culture extracts of Candida parapsilosis and Trichophyton spp. against Trichophyton rubrum.

Onychomycosis is a nail infection caused by dermatophytes, non-dermatophyte fungi, and yeasts, especially Candida species. The present study evaluated the combinatorial effect of different cultured extracts of Candida parapsilosis and Trichophyton mentagrophytes and Trichophyton rubrum with fluconazole, itraconazole, and terbinafine against clinical isolates of Trichophyton rubrum. In addition, investigation of the action of the extracts on the wall or membrane was performed. Pure and mixed cultures of Candida parapsilosis and dermatophytes were filtered through a 0.2-µm membrane and submitted to liquid-liquid extraction using ethyl acetate. After a checkerboard, trial with drugs was performed to evaluate the synergistic interaction with the extract. The results obtained for the minimum inhibitory concentration (MIC) of extracts against the T. rubrum strain in isolation were 500-8000 µg/mL. The MIC range for fluconazole, itraconazole, and terbinafine were 2-32 µg/mL, 0.25-0.5 µg/mL, 0.03-64 µg/mL, respectively. However, when the extract was combined with drugs, the MIC values decreased: extracts 1.9-1000 µg/mL, fluconazole 0.25-4, itraconazole 0.03-0.06 µg/mL, and terbinafine 0.001-0.02 µg/mL. The MIC values of the extracts in the Roswell Park Memorial Institute 1640 medium (RPMI) supplemented with sorbitol did not change, suggesting any action on the cell wall. However, in the presence of RPMI supplemented with ergosterol, MIC values of the extracts increased by up to 2×, indicating action on the fungal cell membrane. A synergistic action was observed between products and drugs, detecting a decrease in MIC values. There is potential and a new therapeutic perspective for fungal control.

Use of Ozonized Water in the Prevention of Surgical Site Infection in Children Undergoing Cardiovascular Surgery

ABSTRACT Introduction: Since the reduction of healthcare-associated infections has been a focus for quality patient care, this study aimed to evaluate the surgical site infection rate of children who underwent cardiovascular surgery after implementation of ozonized water system for hand and body hygiene allied to previously implemented preventive measures. Methods: Two uniformly comparable groups of pediatric patients underwent cardiovascular surgery. Group A (187) patients were operated prior to installation of ozonized water system (March 1 to August 31, 2019), and group B (214) patients were operated after installation of ozonized water system (October 1, 2019, to March 31, 2020). Ozonized water was used for professional hand hygiene and patient body hygiene. Results: There was statistical significance for surgical site infection reduction in group B (P=0.0289), with a relative risk of 0.560 (95% confidence interval = 0.298 to 0.920), inferring the risk of being diagnosed with surgical site infections in group B was 44% less than in group A. There was no statistical significance regarding mechanical ventilation time (P=0.1998) or mortality (P=0.4457). Conclusion: Ozonized water for professional hand hygiene and patient body hygiene was an adjuvant combined with traditional preventive methods to reduce the risk of surgical site infection, although no impact on hospital stay or mortality was observed.

Ultraviolet-C Light-emitting Device Against Microorganisms in Beauty Salons.

Background: Ultraviolet light in the UV-C band is also known as germicidal radiation, and it is widely used for decontamination and disinfection of environments, water, and food. The ultraviolet source transfers electromagnetic energy from a mercury arc lamp to an organism's genetic material. When UV radiation penetrates the cell wall of an organism, it destroys the cell's ability to reproduce, through a physical and not chemical process. Thus, the objective of this study was to evaluate the antimicrobial potential of a new UV-C generating device (Asepsis) against clinically important microorganisms that may be present in beauty centers. Methods: We present here a set of tests performed on tools easy to find in beauty salons (hair-brushes, nail pliers, makeup brushes, and, due to the recent COVID-19 pandemic, face mask samples). They were individually contaminated with bacteria (Pseudomonas aeruginosa, Staphylococcus aureus), fungi (Microsporum canis, Trichophyton rubrum, Candida albicans, Malassezia furfur), and the Chikungunya virus. Different times of exposure were evaluated (1, 3, and 5 minutes). Results: There was notable reduction in the microbial load in every test, in comparison with control groups. Best results were observed on face mask samples, while the makeup brush showed less reduction, even with longer periods of exposure. Conclusions: Beauty salons present a risk of infections due to microbial exposure. The device tested can efficiently inactivate, in a short time, microorganisms contaminating most tools found in this setting. The device also showed promising results against enveloped virus.

Clinical and epidemiological aspects of feline sporotrichosis caused by Sporothrix brasiliensis and in vitro antifungal susceptibility.

Sporotrichosis is a subcutaneous mycosis resulting from the traumatic implantation of pathogenic Sporothrix species. In Brazil, zoonotic transmission plays an important role in the epidemiology of the disease, involving especially cats. The objective of this study was to isolate Sporothrix spp. from cats with signs of sporotrichosis, determining the causative species, clinical and epidemiological aspects, and the in vitro susceptibility profile of the isolates against antifungal drugs. From September 2017 to February 2019, 245 samples of lesions were collected from symptomatic cats in São José do Rio Preto, Brazil. Identification of the isolates was performed by morphophysiological parameters and species-specific polymerase chain reaction. The susceptibility profile of the isolates was determined for five drugs (amphotericin B, itraconazole, ketoconazole, potassium iodide and terbinafine), using the broth microdilution method. Clinical and epidemiological aspects were analyzed based on data contained on investigation forms filled by the veterinarians at moment of collection. Sporothrix spp. were isolated in 189 (77.2%) of the samples. Phenotypic and molecular analyses revealed S. brasiliensis as the only causative agent. In vitro susceptibility testing showed lower MIC values for terbinafine (MIC = 0.03-2 µg/ml), ketoconazole (MIC = 0.03-2 µg/ml), and itraconazole (MIC = 0.03-4 µg/ml). Most of the animals were male (73.5%), adults (96.3%), stray (53.5%), and uncastrated (69.8%). Our results show the expansion of the S. brasiliensis epidemic to an area nearly 840 km apart from the epicenter of the long-lasting outbreak of cat-transmitted sporotrichosis in Rio de Janeiro.

Phenotypic and genotypic profile of the antimicrobial resistance of bacterial isolates and evaluation of physical and chemical potability indicators in groundwater in Brazil.

The aquatic environment has received increasing attention regarding the evolution of bacterial resistance, either as a source of resistance genes or as a matrix for the dissemination of these genes. We evaluated the physicochemical, microbiological and antimicrobial resistance characteristics of 160 samples from alternative water well solutions. According to Ordinance 2914/2011 - MS, 44 (27.5%) samples were considered unsafe if at least one physicochemical parameter exceeded permissible limits. Escherichia coli were found in 30.6% of the unregistered housing estates (UHE) and 1.9% of the local sanitary surveillance system (RW). The total of 158 bacterial strains were isolated from 13 (25%) RW and 68 (63%) UHE, 132 of which (83.5%) were obtained from UHE samples. In the investigation of resistance genes, tetA, qnrS and qnrB genes were detected in three, one and eight isolates, respectively. Our results emphasize the importance of constant surveillance and control of the quality of water supplies.

Ação antimicrobiana do gás ozônio em superfícies e na aeromicrobiota / Acción antimicrobiana del gas ozono en superfi cies y en la microbiota del aire / Antimicrobial action of ozone gas on surfaces and in the air

Resumo Objetivo: Avaliar a ação antimicrobiano do gás ozônio (O3) em superfícies e ar ambiente climatizado artificialmente. Métodos: Estudo experimental/laboratorial e transversal realizado em dez salas de um laboratório de pesquisa em microbiologia médica, com risco de segurança biológica classe 2. As superfícies demarcadas do chão, parede e bancada foram avaliadas, quanto à presença ou ausência de micro-organismos, a partir de coletas feitas com swab umedecido em água destilada estéril, antes e após a exposição do gás O3 gerado por dois equipamentos distintos. Após este procedimento, o swab foi inoculado na superfície do meio de cultura Brain Heart Infusion Agar DIFCO® (BHI), seguindo-se a incubação a 35ºC por 24 horas. Para a análise microbiológica do ar, uma placa com BHI foi exposta aberta por uma hora, antes e após o tratamento do gás O3, sendo incubadas segundo os mesmos critérios. Resultados: A atividade antimicrobiana do gás O3 gerado por ambos os equipamentos foi constatada para todas as áreas investigadas, com registros de redução do número de Unidades Formadoras de Colônias. O potencial de inibição antimicrobiana dos aparelhos se manteve próximo para os critérios de análise adotados, com destaque para as áreas de chão e bancada. Considerando-se todas as salas e percentuais de inibição microbiana, frente aos dois equipamentos, os resultados foram: chão (100%), bancada (90%), parede (50%) e ar, 70%. Conclusão: Os equipamentos geradores de gás O3 apresentaram potencial antimicrobiano para medida de controle de microrganismos presentes em superfícies e ar ambiente climatizado artificialmente, sendo um sanitizante factível para utilização.

Resumen Objetivo: Evaluar la acción antimicrobiana del gas ozono (O3) en superficies y en el aire interior climatizado artificialmente. Métodos: Estudio experimental/de laboratorio y transversal realizado en diez salas de un laboratorio de investigación en microbiología médica, con riesgo de seguridad biológica clase 2. Se evaluaron las superficies delimitadas en el piso, pared y mesa en cuanto a la presencia o ausencia de microorganismos, a partir de muestras recolectadas con hisopo humedecido en agua destilada estéril, antes y después de la exposición del gas O3 generado por dos máquinas distintas. Luego de este procedimiento, el hisopo fue inoculado en la superficie del medio de cultivo Brain Heart Infusion Agar DIFCO® (BHI), y después incubado a 35 °C por 24 horas. Para el análisis microbiológico del aire, se expuso una placa con BHI abierta durante una hora, antes y después del tratamiento del gas O3, y luego se incubó con los mismos criterios. Resultados: Se constató la actividad antimicrobiana del gas O3 generado por ambas máquinas en todas las áreas investigadas, y se registró una reducción del número de unidades formadoras de colonias. El potencial de inhibición antimicrobiana de los dispositivos se mantuvo próximo a los criterios de análisis adoptados, con énfasis en el área del piso y mesa. Considerando todas las salas y porcentajes de inhibición microbiana, con las dos máquinas, los resultados fueron: piso (100 %), mesa (90 %), pared (50 %) y aire (70 %). Conclusión: Las máquinas generadoras de gas O3 presentaron potencial antimicrobiano como medida de control de microorganismos presentes en superficies y aire interior climatizado artificialmente, lo que lo convierte en un desinfectante factible para ser usado.

Abstract Objective: Assess the antimicrobial action of ozone gas (O3) on surfaces and artificially cooled ambient air. Methods: Cross-sectional experimental/laboratory study carried out in ten rooms of a medical microbiology research lab, with class 2 biosafety risk. The demarcated surfaces on the floor, wall and counter were assessed in relation to the presence or absence of microorganisms, based on collections done with swabs dampened in sterile distilled water, before and after exposure to ozone gas produced by two different generators. After this procedure, each swab was inoculated on the surface of a Brain Heart Infusion Agar DIFCO® (BHI) culture, followed by incubation at 35ºC for 24 hours. For the microbiological analysis of the air, a petri dish with BHI was openly exposed for one hour, before and after treatment with O3 gas, and were incubated according to the same criteria. Results: The antimicrobial activity of the O3 gas produced by both generators was checked in all the areas investigated, with records indicating a decrease in the number of colony-forming units. The antimicrobial inhibition potential of the generators was close to the analysis criteria adopted, particularly for the floor and counter areas. Based on all the rooms and microbial inhibition percentages, in relation to the two generators, the results were: floor (100%), counter (90%), wall (50%) and air (70%). Conclusion: The O3 generators had antimicrobial potential as a procedure for controlling microorganisms present on surfaces and in artificially cooled ambient air, constituting a feasible sanitizer.

Broad Diversity of Fungi in Hospital Water.

INTRODUCTION: Some studies have reported the occurrence of microorganisms isolated from water. Considering these microorganisms, fungi are known to occur ubiquitously in the environment, including water, and some are pathogenic and may cause health problems, especially in immunocompromised individuals. The aim of this study was to identify fungi in hospital water samples and to correlate their presence with the concentration of free residual chlorine. METHODS: Water samples (100 mL) were collected from taps (n = 74) and water purifiers (n = 14) in different locations in a university hospital. Samples were filtered through a nitrocellulose membrane and placed on Sabouraud dextrose agar and incubated for 24 hours at 30°C. Fungi were identified according to established methods based on macroscopic and microscopic characteristics (filamentous) and physiological tests (yeasts). Free chlorine residual content was measured at the time of sample collection. RESULTS: Seventy species of fungi were identified in the water samples and about 56% of the water samples contained culturable fungi. Cladosporium oxysporum, Penicillium spinulosum, and Aspergillus fumigatus were the most common filamentous fungi. Aureobasidium pullulans and Candida parapsilosis were the most common yeasts. Chemical analyses revealed that free residual chlorine was present in 81.8% of the samples within recommended concentrations. Among samples from water purifiers, 92.9% showed low levels of free residual chlorine (<0.2 mg/L). There was no significant association between chlorine concentrations (either within or outside the recommended range) and the presence of filamentous fungi and yeasts. CONCLUSIONS: This study showed that hospital water can be a reservoir for fungi, some of which are potentially harmful to immunocompromised patients. Free residual chlorine was ineffective in some samples.

Comparing activity, toxicity and model membrane interactions of Jelleine-I and Trp/Arg analogs: analysis of peptide aggregation.

Increasing resistance in antibiotic and chemotherapeutic treatments has been pushing studies of design and evaluation of bioactive peptides. Designing relies on different approaches from minimalist sequences and endogenous peptides modifications to computational libraries. Evaluation relies on microbiological tests. Aiming a deeper understanding, we chose the octapeptide Jelleine-I (JI) for its selective and low toxicity profile, designed small modifications combining the substitutions of Phe by Trp and Lys/His by Arg and tested the antimicrobial and anticancer activity on melanoma cells. Biophysical methods identified environment-dependent modulation of aggregation, but critical aggregation concentrations of JI and analogs in buffer show that peptides start membrane interactions as monomers. The presence of model membranes increases or reduces the partial aggregation of peptides. Compared to JI, analog JIF2WR shows the lowest tendency to aggregation on bacterial model membranes. JI and analogs are lytic to model membranes. Their composition-dependent performance indicates preference for the higher charged anionic bilayers in line with their superior performance toward Staphylococcus aureus and Streptococcus pneumoniae. JIF2WR presented the higher partitioning, higher lytic activity and lower aggregated contents. Despite these increased membranolytic activities, JIF2WR exhibited comparable antimicrobial activity in relation to JI at the expenses of some loss in selectivity. We found that the substitution Phe/Trp (JIF2W) tends to decrease antimicrobial but to increase anticancer activity and aggregation on model membranes and the toxicity toward human cells. However, the concomitant substitution Lys/His by Arg (JIF2WR) modulates some of these tendencies, increasing both the antimicrobial and the anticancer activity while decreasing the aggregation tendency.

Epidemiological profile of patients with congenital and gestational syphilis in a city in the State of São Paulo, Brazil / Perfil epidemiológico de pacientes com sífilis congênita e gestacional em um município do Estado de São Paulo, Brasil

Abstract Objectives: to assess the epidemiological profile of congenital and syphilis during pregnancy in residents of São José do Rio Preto in São Paulo State. Methods: ecological study of the epidemiological profile of patients with congenital and gestational syphilis, based on the Sistema de Informação de Agravos de Notificação (Information System for Notifiable Diseases) from 2007 to 2016. Results: there were 396 cases of syphilis reported in pregnant women and 290 of congenital syphilis. In 2016, the rate of detecting syphilis in pregnant women was 13.2 cases/1,000 live births, while congenital syphilis the incidence rate was 6.5 cases/1,000 live births. For gestational syphilis, 54% of the diagnosis was performed in 2nd or 3rd trimester and 85% were reported at the primary care. Adequate treatment for pregnant women occurred in 96% of the notifications with 52% of partners treated. In congenital syphilis, 82% of the mothers underwent prenatal care. However, 94% of the pregnant women were treated inadequately while 82% of the partners did not receive any treatment. Conclusions: there has been an increase in the number of cases of gestational syphilis in pregnant women and a decrease in the cases of congenital syphilis from 2014. These results showed that the goal of 0.5 case/1,000 live births proposed by World Health Organization is still far from being achieved in this city.

Resumo Objetivos: conhecer o perfil epidemiológico da sífilis congênita e em gestantes nos residentes de São José do Rio Preto/SP. Métodos: estudo ecológico do perfil epidemiológico dos pacientes com sífilis congênita e gestacional, a partir de dados coletados no Sistema de Informação de Agravos de Notificação entre 2007 e 2016. Resultados: foram notificados 396 casos de sífilis em gestantes e 290 de sífilis congênita. Em 2016, a taxa de detecção da sífilis em gestantes foi 13,2 casos/1.000 nascidos vivos, enquanto a sífilis congênita, a taxa de incidência foi 6,5 casos/1.000 nascidos vivos. Para sífilis gestacional, 54% do diagnóstico foram realizados no 2° ou 3° trimestre e 85% notificadas na atenção primária. O tratamento adequado das gestantes ocorreu em 97% das notificações, com 52% dos parceiros tratados. Na sífilis congênita, 82% das mães realizaram o pré-natal, entretanto, 94% das gestantes foram tratadas inadequadamente e 82% dos parceiros não realizaram o tratamento. Conclusões: foi observado o aumento no número de casos de sífilis em gestantes e uma queda dos casos de sífilis congênita a partir de 2014. Com esses resultados, é notório que a meta de 0,5 caso/1.000 nascidos vivos proposta pela Organização Mundial da Saúde ainda está distante de ser alcançada no município.

Wide diversity of fungal species found in wellwater for human consumption: an analytical cross-sectional study

ABSTRACT BACKGROUND: Fungi are ubiquitous in the environment. They are able to grow in water and many of them may be opportunistic pathogens. OBJECTIVE: The aims were to identify fungi in registered wells (RWs) and nonregistered wells (NRWs) that tap into groundwater; and to correlate the results from physicochemical assays on this water (free residual chlorine and pH) with the presence of fungi. DATA AND SETTING: Analytical cross-sectional quantitative study on groundwater wells in São José do Rio Preto, São Paulo, Brazil. METHODS: 52 samples of 500 ml of water were collected from RWs and 107 from NRWs. These were sent to a microbiology laboratory to identify any fungi that were present. In addition, free residual chlorine and pH were measured immediately after sample collection. Several statistical analysis tests were used. RESULTS: Fungal contamination was present in 78.8% of the samples from RWs and 81.3% from NRWs. Filamentous fungi were more prevalent than yeast in both types of wells. There was no significant difference in presence of fungi according to whether chloride and pH were within recommended levels in RWs; or according to whether pH was within recommended levels in NRWs. Furthermore, there was no statistical difference in the levels of fungal contamination between RWs and NRWs. CONCLUSION: Both RWs and NRWs are potential reservoirs for many types of fungi. Many of these may become opportunistic pathogens if they infect immunosuppressed individuals. Furthermore, this study confirms that fungi are able to grow even when chlorine and pH parameters are within the standards recommended.

Evaluation of disinfection of surfaces at an outpatient unit before and after an intervention program.

BACKGROUND: Cleaning and disinfection processes must be improved so that there is a reduction in environmental contamination of frequent-contact surfaces. The objective of this study was to evaluate cleaning and disinfection of surfaces at a specialized healthcare unit after an intervention program. METHODS: Exploratory, longitudinal, and correlational study carried out in a medium-complexity clinic. Two hundred and forty samples from five surfaces were collected during three phases: diagnosis; implementation of an intervention program; and evaluation of immediate and long-term effects. In total, 720 evaluations were made, performed through three monitoring methods: visual inspection; adenosine triphosphate bioluminescence assay (ATP); and aerobic colony count (ACC). The Wilcoxon, Mann-Whitney, and Fisher's Exact tests were run to analyze data statistically. RESULTS: Cleaning and disinfection of surfaces were not being performed properly in most cases. Failure rates of surfaces reached 37.5 and 100% when the ATP and ACC procedures were used, respectively. However, after an intervention program, an improvement occurred. Success rates increased by 43.96% (ATP) and 12.46% (ACC) in phase I, by 70.6% (ATP) and 82.3% (ACC) immediately after interventions, and by 76.52% (ATP) and 85.76% (ACC) two months after the changes, showing that the program was effective. CONCLUSION: The present study reveals that implementing intervention actions with a cleaning and healthcare team brings benefits to prevent the spread of pathogenic agents through frequently touched hospital surfaces.

Intravaginal Delivery of Syngonanthus nitens (Bong.) Ruhland Fraction Based on a Nanoemulsion System Applied to Vulvovaginal Candidiasis Treatment.

In this study, was evaluated the chemical composition of a fraction from Syngonanthus nitens extract and its antimicrobial potential unloaded (Fr3) and loaded (F9Fr3) into a nanoemulsion (F9) composed of cholesterol as the oil phase (10%), polyoxyethylene 20-cetyl ether and soy phosphatidylcholine (2:1) as surfactant (20%), and a solution of phosphate buffer (pH 7.4) plus chitosan polymer dispersion (0.25%) as the aqueous phase (70%) to use for VVC treatment. Phytochemical procedures showed that Fr3 is rich in luteolin, which is responsible for the antimicrobial activity. F9 development showed satisfactory parameters for use in the vulvovaginal candidiasis (VVC) treatment, as F9 demonstrated pseudoplastic, elastic behavior, and adhesive properties on vaginal mucosa. In addition, we observed improvement in antimicrobial potential of Fr3 on planktonic and biofilms after incorporation in F9. Fr3 and F9Fr3 showed satisfactory parameters related to toxic profiles in cell lines and in a model of acute toxicity by Artemia salina. The in vivo VVC assay showed that F9Fr3 was more active than unloaded Fr3 in VVC treatment. In conclusion, this work showed that use of a fraction rich in luteolin can be a used as an antimicrobial for treatment of vaginal infections and that use of nanostructured lipid systems was an important factor in the biological efficacy of Fr3, especially in treatment of acute VVC.

Wide diversity of fungal species found in wellwater for human consumption: an analytical cross-sectional study.

BACKGROUND: Fungi are ubiquitous in the environment. They are able to grow in water and many of them may be opportunistic pathogens. OBJECTIVE: The aims were to identify fungi in registered wells (RWs) and nonregistered wells (NRWs) that tap into groundwater; and to correlate the results from physicochemical assays on this water (free residual chlorine and pH) with the presence of fungi. DATA AND SETTING: Analytical cross-sectional quantitative study on groundwater wells in São José do Rio Preto, São Paulo, Brazil. METHODS: 52 samples of 500 ml of water were collected from RWs and 107 from NRWs. These were sent to a microbiology laboratory to identify any fungi that were present. In addition, free residual chlorine and pH were measured immediately after sample collection. Several statistical analysis tests were used. RESULTS: Fungal contamination was present in 78.8% of the samples from RWs and 81.3% from NRWs. Filamentous fungi were more prevalent than yeast in both types of wells. There was no significant difference in presence of fungi according to whether chloride and pH were within recommended levels in RWs; or according to whether pH was within recommended levels in NRWs. Furthermore, there was no statistical difference in the levels of fungal contamination between RWs and NRWs. CONCLUSION: Both RWs and NRWs are potential reservoirs for many types of fungi. Many of these may become opportunistic pathogens if they infect immunosuppressed individuals. Furthermore, this study confirms that fungi are able to grow even when chlorine and pH parameters are within the standards recommended.

Biologia Molecular como ferramenta de detecção fúngica no sangue: auxílio diagnóstico e redução de gastos / Molecular Biology as a tool for fungal detection on blood: diagnostic aid and cost reduction

Introdução: Os avanços médicos das últimas décadas contribuíram para aumentar a sobrevida de pacientes críticos e com a resposta imune comprometida. Consequentemente, a população em risco de adquirir infecções de origem fúngica também cresceu. Com altas taxas de morbidade e mortalidade, o difícil diagnóstico deste tipo de infecção, em conjunto com terapias ineficazes, gera elevados custos e sobrecarga ao sistema de saúde. Objetivos: Padronizar um método molecular de detecção fúngica diretamente do sangue e avaliar esta técnica comparativamente com a atualmente considerada padrão-ouro (hemocultura), associando aspectos clínicos, tempo de realização das técnicas e os custos envolvidos. Casuística e Métodos:Neste sentido, 94 pacientes com suspeita de infecção de corrente sanguínea foram submetidos a uma técnica de nestedPCR para detecção de DNA fúngico. Resultados: A técnica molecular foi positiva em 48,9% das amostras, enquanto que a hemocultura foi positiva em apenas 13,0% dos casos. Esses resultados demonstram uma alta sensibilidade do nested PCR e com um valor preditivo negativo de 100% em pacientes com suspeita clínica de infecção fúngica sistêmica e em situações de risco. O tempo de realização do método e os custos associados a ele, em comparação à hemocultura, também demonstraram seu potencial para uso clínico. Conclusões: Em comparação com a hemocultura, o método padronizado de nestedPCR constitui um teste rápido e economicamente viável capaz de descartar uma infecção sistêmica provocada por fungo, podendo facilitar o diagnóstico e evitar terapias ineficientes e caras, diminuir o tempo de internação e os impactos econômicos gerados por esse tipo de infecção.

Introduction: Medical advances in the past decades have contributed to the increase of survival of critically ill patients and the ones with impaired immune response. Consequently, the population at risk of acquiring a fungal infection also has increased. This type of infection generates expensive costs and heavy burden to the Health system. It also brings high morbidity and mortality rates, difficulty in diagnosing, and ineffective therapies. Objectives: Standardize a molecular method of fungal detection directly from blood and compare this technique with the blood culture, which is currently considered the gold standard. It associates clinical aspects, time, and costs involved. Patients and Methods: In this sense, 94 patients with suspected bloodstream infection were submitted to the technique of nested PCR for detection of fungal DNA. Results: The molecular technique was positive in 48.9% of the samples, while the blood culture was positive in only 13% of the cases. These results demonstrate high sensibility of the nested PCR and negative predictive value of 100%. The performing time and the costs associated with the method also demonstrated its value for clinical use. Conclusions: Therefore, the nested PCR is a quick and economically viable test, capable of ruling out a systemic infection caused by fungus, being able to facilitate the diagnosis, avoid inefficient and expensive therapies, and decrease the length of hospital stay, reducing the burden caused by this type of infection.

Microbiological analysis of contact lens cases: impact of the hospital environment / Análise microbiológica de estojos de lentes de contato: impacto do ambiente nosocomial

ABSTRACT Purpose: The aim of this study was to analyze the bacterial and fungal microbiota found in contact lens cases among two groups of patients to correlate the data on the habits of contact lens users and to evaluate whether there is a difference in the culture results between users of ridged and nonridged contact lens cases. Methods: Two groups (35 patients per group) were included, consisting of hospital workers and those who had not visited a hospital in the past 30 days, and a questionnaire regarding epidemiological data and habits related to contact lens and lens case use was administered. In addition, 140 samples collected from the right and left compartments of each lens case by swabbing the bottom of the wells were tested using bacterioscopy as well as fungal and bacterial cultures via computerized identification of the species. Results: No fungal growth was identified in any of the 70 contact lens cases; however, bacteria were found in 39 cases, and there was no statistical difference between the groups. Most bacteria (>85%) were gram-negative bacilli. Contamination inone compartment of the contact lens case elevated the risk of contamination of the other side (>80%). Moreover, contamination was statistically higher in the ridged cases than in nonridged cases (p=0.0149). Conclusion: The types of bacteria contaminating the cases are generally not seen in eye diseases associated with contact lens use, suggesting that other decisive variables are involved in eye infection from a contaminated lens or case. Fungal contamination of contact lens cases appears to be an exception. Ridged cases are commonly used by contact lens wearers and present a potential risk to eye health. In addition, the results of bacterial tests between hospital workers and those who did not visit a hospital were not significantly different.

RESUMO Objetivo: O objetivo deste trabalho foi analisar a microbiota bacteriana e fúngica encontrada em estojos de lentes de contato em dois grupos, correlacionar os dados sobre os hábitos de uso de lentes de contato e avaliar se há diferença na positividade das culturas entre os usuários estojos de lentes de contato com ranhuras e sem ranhuras. Métodos: Dois grupos foram formados, trabalhadores do hospital e pessoas que não visitaram o hospital (35 indivíduos por grupo), e um questionário foi aplicado sobre dados epidemiológicos e hábitos relacionados ao uso de lentes de contato e estojos de lentes. Além disso, 140 amostras, coletadas do compartimento direito e esquerdo de cada estojo de lente, esfregando o fundo dos mesmos, foram testadas por bacterioscopia e por culturas de fungos e bactérias, com identificação computadorizada da espécie. Resultados: Não houve crescimento fúngico em nenhum dos 70 estojos de lentes de contato, porém bactérias foram encontradas em 39; não houve diferença estatística entre os grupos. A maioria das bactérias (>85%) eram bacilos gram-negativos. Quando um compartimento estava contaminado, o risco de contaminação do outro compartimento era elevado (>80%). A contaminação foi estatisticamente maior nos estojos com ranhuras (p=0,0149). Conclusão: A contaminação dos estojos parece ocorrer por bactérias que, em geral, não são encontradas em doenças oculares associadas ao uso de lentes de contato, sugerindo que existem outras variáveis decisivas nas infecções oculares de uma lente ou estojo contaminado. Contaminação de estojos de lentes de contato com fungos parece ser uma exceção. O uso de estojos com ranhuras é uma prática comum e apresenta um risco potencial à saúde ocular. Não foram encontradas diferenças significativas nos resultados dos testes bacterianos entre trabalhadores hospitalares e pessoas que não visitaram o hospital.

Microbiological analysis of contact lens cases: impact of the hospital environment.

PURPOSE: The aim of this study was to analyze the bacterial and fungal microbiota found in contact lens cases among two groups of patients to correlate the data on the habits of contact lens users and to evaluate whether there is a difference in the culture results between users of ridged and nonridged contact lens cases. METHODS: Two groups (35 patients per group) were included, consisting of hospital workers and those who had not visited a hospital in the past 30 days, and a questionnaire regarding epidemiological data and habits related to contact lens and lens case use was administered. In addition, 140 samples collected from the right and left compartments of each lens case by swabbing the bottom of the wells were tested using bacterioscopy as well as fungal and bacterial cultures via computerized identification of the species. RESULTS: No fungal growth was identified in any of the 70 contact lens cases; however, bacteria were found in 39 cases, and there was no statistical difference between the groups. Most bacteria (>85%) were gram-negative bacilli. Contamination inone compartment of the contact lens case elevated the risk of contamination of the other side (>80%). Moreover, contamination was statistically higher in the ridged cases than in nonridged cases (p=0.0149). CONCLUSION: The types of bacteria contaminating the cases are generally not seen in eye diseases associated with contact lens use, suggesting that other decisive variables are involved in eye infection from a contaminated lens or case. Fungal contamination of contact lens cases appears to be an exception. Ridged cases are commonly used by contact lens wearers and present a potential risk to eye health. In addition, the results of bacterial tests between hospital workers and those who did not visit a hospital were not significantly different.

Efficiency of cleaning and disinfection of surfaces: correlation between assessment methods / Eficiencia de la limpieza y desinfección de superficies: correlación entre métodos de evaluación / Eficiência da limpeza e desinfecção de superfícies: correlação entre métodos de avaliação

ABSTRACT Objective: to assess the correlation among the ATP-bioluminescence assay, visual inspection and microbiological culture in monitoring the efficiency of cleaning and disinfection (C&D) of high-touch clinical surfaces (HTCS) in a walk-in emergency care unit. Method: a prospective and comparative study was carried out from March to June 2015, in which five HTCS were sampled before and after C&D by means of the three methods. The HTCS were considered dirty when dust, waste, humidity and stains were detected in visual inspection; when ≥2.5 colony forming units per cm2 were found in culture; when ≥5 relative light units per cm2 were found at the ATP-bioluminescence assay. Results: 720 analyses were performed, 240 per method. The overall rates of clean surfaces per visual inspection, culture and ATP-bioluminescence assay were 8.3%, 20.8% and 44.2% before C&D, and 92.5%, 50% and 84.2% after C&D, respectively (p<0.001). There were only occasional statistically significant relationships between methods. Conclusion: the methods did not present a good correlation, neither quantitative nor qualitatively.

RESUMEN Objetivo: evaluar correlación del test ATP-bioluminiscencia con inspección visual y cultivo microbiológico en monitoreo de eficiencia de limpieza y desinfección (L&D) de superficies clínicas altamente tocadas (SCAT) en unidad de pronta atención. Métodos: estudio comparativo, prospectivo, realizado de marzo a junio de 2015, cuando cinco SCAT fueron muestreadas antes y después de L&D de rutina por los tres métodos. Las SCAT fueron consideradas sucias cuando presentaban: en inspección visual: polvo, deyecciones, humedad y manchas; en cultivo: ≥205 unidades formadoras de colonias por cm2, y en ATP-bioluminiscencia: ≥5 Unidades Relativas de Luz por cm2. Resultados: fueron realizadas 720 evaluaciones, 240 por método. La tasa global de superficies limpias por inspección visual, cultivo y ATP-bioluminiscencia fue, respectivamente, 8,3%, 20,8% y 44,2% antes de la L&D y de 92,5%, 50% y 84,2% después (p<0,001). Existieron sólo asociaciones puntuales estadísticamente significativas entre los métodos. Conclusión: los métodos no presentan buena correlación cuantitativa, ni cualitativa.

RESUMO Objetivo: avaliar a correlação do teste de ATP-bioluminescência com inspeção visual e cultura microbiológica na monitorização da eficiência da limpeza e desinfecção (L&D) de superfícies clínicas altamente tocadas (SCAT) em unidade de pronto atendimento. Métodos: estudo comparativo, prospectivo, conduzido de março a junho de 2015, de forma que cinco SCAT foram amostradas antes e depois da L&D de rotina pelos três métodos. As SCAT foram consideradas sujas quando apresentaram: na inspeção visual, poeira, dejetos, umidade e manchas; na cultura, ≥2,5 unidades formadoras de colônias por cm2 e; no ATP-bioluminescência, ≥5 Unidades Relativas de Luz por cm2. Resultados: foram realizadas 720 avaliações, sendo 240 por método. A taxa global de superfícies limpas por inspeção visual, cultura e ATP-bioluminescência foi, respectivamente, de 8,3%, 20,8% e 44,2% antes da L&D e de 92,5%, 50% e 84,2% após (p<0,001). Houve apenas associações pontuais estatisticamente significativas entre os métodos. Conclusão: os métodos nem apresentaram boa correlação quantitativa, nem, qualitativa.

Efficiency of cleaning and disinfection of surfaces: correlation between assessment methods.

OBJECTIVE: to assess the correlation among the ATP-bioluminescence assay, visual inspection and microbiological culture in monitoring the efficiency of cleaning and disinfection (C&D) of high-touch clinical surfaces (HTCS) in a walk-in emergency care unit. METHOD: a prospective and comparative study was carried out from March to June 2015, in which five HTCS were sampled before and after C&D by means of the three methods. The HTCS were considered dirty when dust, waste, humidity and stains were detected in visual inspection; when ≥2.5 colony forming units per cm2 were found in culture; when ≥5 relative light units per cm2 were found at the ATP-bioluminescence assay. RESULTS: 720 analyses were performed, 240 per method. The overall rates of clean surfaces per visual inspection, culture and ATP-bioluminescence assay were 8.3%, 20.8% and 44.2% before C&D, and 92.5%, 50% and 84.2% after C&D, respectively (p<0.001). There were only occasional statistically significant relationships between methods. CONCLUSION: the methods did not present a good correlation, neither quantitative nor qualitatively.

Effectiveness of ATP bioluminescence assay for presumptive identification of microorganisms in hospital water sources.

BACKGROUND: Laboratory analysis of organisms in water include arduous methods, such as the multiple tube and membrane filter. The ATP bioluminescence system, proposes a new way of measuring cellular material in water by measuring adenosine triphosphate (ATP) levels, which are expressed in relative light units (RLU). The ATP bioluminescence assay has been increasingly used to assess the microbiological safety of the hospital environment. However, there are few studies investigating the use of this methodology to evaluate the microbiological quality of water. The objective of the present study was to verify whether ATP, as measured by the 3 M™ Clean-Trace Water™ ATP test, can be used as an alternative tool for presumptive testing for the presence of microorganisms in hospital water. METHODS: Water samples (N = 88) were collected from faucets (74) and water purifiers (14) in a university hospital. The sample were filtered by the membrane filter technique (100 mL for bacterial analysis and 100 mL for fungal analysis) and then submitted to ATP bioluminescence assay to the determine quantity of RLU in each sample. In order to compare RLU and the presence of microorganisms, a receiver operating characteristic (ROC) curve was used to calculate sensitivity and specificity (levels higher than 90% were considered significant). In addition, control tests were conducted to compare RLU to the quantities of bacterial and fungal organisms added to distilled water (ANOVA and Tukey's tests; p ≤ 0.05). This inoculum was compared to RLU emission, and the data were analyzed by calculating the Pearson's correlation coefficient, with a 95% confidence interval. RESULTS: In the present study, 94.3% of the water samples presented bacterial growth. Of these, 15.6% showed heterotrophic bacteria above recommended levels and fungal contamination was detected in 55.6% of samples. Sensitivity and specificity of the samples were not significant (< 90%), and the correlation between ATP and the presence of these microorganisms in the samples (hospital water) was not significant, whereas, in distilled water, the results revealed a significant difference (p < 0.0001). CONCLUSIONS: These results demonstrated that the ATP test cannot be used as an alternative tool for presumptive assessment of the presence of microorganisms in water.