Antifungal agents incorporated in denture base materials: a scoping review of the current evidence and technology prospecting.

BACKGROUND: The aim of this study was to summarize existing data and perform technological prospecting on the effect of incorporating antifungal agents into denture base materials in inhibiting Candida spp., as well as to explore the antimicrobial properties of these materials. METHODS: A comprehensive electronic search was carried out in six major bibliographic databases (PubMed, Scopus, Embase, Cochrane Library, Web of Science and Lilacs) until February 2024. In addition, international patent databases were also examined. The search process, study and patent selection, data extraction and risk of bias assessment were carried out independently by researchers. The collected data underwent qualitative analysis. RESULTS: A total of 10 718 articles were identified in the searched databases, of which 40 documents were included for qualitative data analysis (articles: 31; patents: 9). The majority of the studies focused on investigating tissue conditioners (n = 14) and acrylic resins (n = 14). The primary antifungal agents studied were nystatin (n = 15) and fluconazole (n = 13). The most commonly utilized microbiological evaluation methodology was the agar diffusion test (n = 16), followed by the microdilution (n = 7) and biofilm formation assays (n = 7). All of the studies investigated the inhibitory effect of these materials against Candida species. CONCLUSION: The incorporation of antifungal agents into denture base materials has been extensively studied and has shown a significant inhibitory response against Candida spp. across various methodological assays.

Antifungal Activity of Morinda citrifolia Methanolic Extract against Candida albicans: An In Vitro Study.

Aim: Natural medicine used as an alternative and/or complementary treatment to counteract diseases is of great importance in public health. Therefore, the purpose of the present study was to assess the in vitro antifungal activity of Morinda citrifolia methanolic extract of peel, pulp, and seed against Candida albicans. Materials and Methods: The present study was experimental in vitro and cross-sectional. Eight replicates were prepared in Sabouraud dextrose agar with five wells each, where 0.12% chlorhexidine, distilled water, and methanolic extract of seed, peel, and pulp of Morinda citrifolia fruit were placed at concentrations of 10,690, 8,270, and 6,430 mg/mL, respectively, to evaluate sensitivity according to Duraffourd's scale. In addition, the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were determined by dilution and agar seeding method. Statistical analysis was performed by analysis of variance (ANOVA) and Tukey's post hoc test, considering a significance level of P < 0.05. Results: The inhibition halos of Morinda citrifolia methanolic extract of seed, peel, and pulp against Candida albicans measured on average 15.94, 11.94, and 11.56 mm, respectively. The MIC of seed, peel, and pulp extract were 1366.25, 2067.5, and 1607.5 mg/mL respectively, whereas the MFC for seed, peel, and pulp extract were 2672.50, 2067.5, and 3215 mg/mL, respectively. Moreover, seed extract presented significantly higher antifungal activity than peel and pulp (P < 0.001). Conclusions: Morinda citrifolia methanolic extract of peel, pulp, and seed showed fungistatic and fungicidal effect against Candida albicans, being this very sensitive to seed extract with a MIC of 1366.25 mg/mL and a MFC of 2672.5 mg/mL, which allows recommending the development of effective pharmacological formulations for the control of candidiasis.

Osteomyelitis by Microsporum canis and Staphylococcus spp. in cat (Felis catus) - case report.

BACKGROUND: Staphylococcus spp and Microsporum canis are zoonotic microorganisms which can cause infections and systemic diseases. The bone infection is usually caused by invasion of pathogen through the hematologic route. Mixed osteomyelitis caused by bacteria and fungi is rare, and to date, there have been no reports of mixed osteomyelitis with Staphylococcus spp. and Microsporum canis. CASE PRESENTATION: This essay reports an atypical presentation of mixed osteomyelitis (Staphylococcus spp. and Microsporum canis) in a domestic cat. A 15-month-old female Persian cat was presented to a veterinary service; the main complaint was the appearance of a nodule in the mandibular ventral rostral region. A radiographic exam performed on the animal showed proliferative and osteolytic bone lesions. The patient was submitted to a biopsy for histopathological evaluation, along with bacterial and fungal cultures. Results showed mixed osteomyelitis by Staphylococcus spp. and Microsporum canis. Microbial Sensitivity Test was performed to choose a more suitable treatment. Two surgical procedures were executed to resect and curette the lesion, and treatments with anti-inflammatory, antibiotic, and antifungal drugs were established, showing a positive clinical evolution. After 8 months of treatment, the patient's owner moved to a different city, and the animal was seen by other veterinarians, who followed along with the same treatment. However, due to complications and a diminishing quality of life over 4 years of diagnosis, the patient was euthanized. CONCLUSION: Given the above, mixed osteomyelitis is difficult to treat and can cause losses of life quality resulting death, especially in infections where M. canis is the agent causing the disease. Bacterial osteomyelitis is more frequently reported. But the lack of investigation of microorganisms other than bacteria, such as fungal cases, may imply in underdiagnosed cases. Treatment of osteomyelitis can be difficult considering the difficulties in isolating the pathological agent, resistance to the drug used, prolonged treatment time, and cost.

Perfil da microbiota e padrões de resistência aos antimicrobianos em uroculturas de pacientes transplantados renais / Profile of the microbiot and anti-microbain resistance patterns in urocultures of reneal transplant patients

Objetivo: analisar o perfil de micro-organismos presentes e resistência destes aos antimicrobianos em uroculturas de pacientes transplantados renais no período de 2021-2022. Métodos: trata-se de um estudo transversal com análise quantitativa dos dados de uroculturas positivas de pacientes transplantados renais, acompanhados no Hospital Geral de Fortaleza entre janeiro de 2021 a dezembro de 2022. Foi empregado um instrumento de pesquisa elaborado, contendo variáveis classificatórias, e os dados foram obtidos por meio de registros das uroculturas existentes no sistema de prontuário eletrônico utilizado pelo hospital. Resultados: das 534 uroculturas solicitadas, 36,7% apresentaram resultado positivo, sendo 60,4% de mulheres com idades entre 20 e 59 anos. A maioria dos casos foram desenvolvidos por pacientes que receberam acompanhamento ambulatorial (56,2%). Os micro-organismos isolados foram, predominantemente, enterobactérias (81,34%), com prevalência de E.coli (69,30%). Os perfis de sensibilidade antimicrobiana variaram, com a resistência da E.coli a antibióticos como ampicilina, ácido nalidíxico, norfloxacino e ciprofloxacino. Conclusões: essas descobertas fornecem informações importantes sobre métodos clínicos específicos, métodos preventivos e melhorias na qualidade de vida dos transplantados renais.

Objective: to analyze the profile of microorganisms present and their resistance to antimicrobials in urocultures of renal transplant patients in 2021-2022. Methods: it is a cross-sectional study with quantitative data analysis from positive urocultures of renal transplant patients accompanied at the General Hospital of Fortaleza between January 2021 and December 2022. An elaborate research instrument containing classification variables was employed, and the data were obtained through records of the urocultures existing in the electronic checkbook system used by the hospital. Results: of the 534 urocultures requested, 36.7% showed a positive result, of which 60.4% were women aged between 20 and 59. Most cases were developed by patients who received outpatient follow-up (56.2%). The isolated microorganisms were predominantly enterobacteria (81.34%), with the prevalence of E.coli (69.30%). Antimicrobial sensitivity profiles varied, with E.coli resistance to antibiotics such as ampicillin, nalidixic acid, norfloxacin, and ciprofloxacin. Conclusion: these findings provide important information about specific clinical methods, preventive methods, and improvements in the quality of life of renal transplant patients.

Anti-Candida activity and in vitro toxicity screening of antifungals complexed with ß-cyclodextrin.

The emergence of resistant fungal species and the toxicity of currently available antifungal drugs are relevant issues that require special consideration. Cyclodextrins inclusion complexes could optimize the antimicrobial activity of such drugs and create a controlled release system with few side effects. This study aimed to assess the in vitro toxicity and antifungal effectiveness of nystatin (Nys) and chlorhexidine (Chx) complexed or not with ß-cyclodextrin (ßCD). First, a drug toxicity screening was performed through the Artemia salina bioassay. Then, the minimum inhibitory concentrations (MICs) against Candida albicans were determined with the broth microdilution test. After MICs determination, the cytotoxicity of the drugs was evaluated through the methyl-thiazolyl-tetrazolium (MTT) and neutral red (NR) assays and through cell morphology analysis. The PROBIT analysis was used to determine the median lethal concentration (LC50), and the cell viability values were submitted to one-way analysis of variance(ANOVA)/Tukey (α = 0.05). Overall, the ßCD-complexed antifungals were less toxic against A. salina than their raw forms, suggesting that inclusion complexes can reduce the toxicity of drugs. The MICs obtained were as follows: Nys 0.5 mg/L; Nys:ßCD 4 mg/L; Chx 4 mg/L; and Chx:ßCD 8 mg/L. Chx showed significant cytotoxicity (MTT: 12.9 ± 9.6%; NR: 10.6 ± 12.5%) and promoted important morphological changes. Cells exposed to the other drugs showed viability above 70% with no cellular damage. These results suggest that antifungals complexed with ßCD might be a biocompatible option for the treatment of Candida-related infections.

Prevalence and antimicrobial resistance profile of pathogens isolated from patients with urine tract infections admitted to a university hospital in a medium-sized Brazilian city

ABSTRACT This study aimed to determine the antibiotic profile of microorganisms isolated from urine samples of patients with community urine tract infections (UTI) admitted to the University Hospital of the Federal University of Sao Carlos to support an appropriate local empirical treatment. A retrospective cross-sectional study was conducted from October 2018 to October 2020. Data from 1,528 positive urine cultures for bacterial pathogens and antibiograms were tabulated. Bacterial species prevalence and their resistance profile were analyzed and compared by sex and age. For Gram-negative fermenting bacteria, resistance rates were compared between patients with previous hospitalization and the total of infections caused by this group. For comparisons, the Chi-square test was performed, using Fisher's exact test when necessary (BioEstat program, adopting p ≤ 0.05). A multivariate analysis was applied to assess the effect of the studied variables in predicting multidrug resistance. Infections were more prevalent in women and older adults. Gram-negative bacteria represented 90.44% of total cultures. In both sexes, E. coli prevalence was significantly higher in adults compared with older adults (p < 0.0001). For several antibiotics, resistance rates were higher in the older adults compared with other ages and in patients with Gram-negative fermenting infections and previous hospitalization compared with the total of infections by this group of bacteria. The closer to the hospitalization, the higher the number of antibiotics with superior resistance rates. Resistance rates for aminoglycosides, carbapenems, ceftazidime, nitrofurantoin, piperacillin+tazobactam, and fosfomycin were less than 20%, considered adequate for empirical treatment. Only hospitalization in the previous 90 days was statistically significant in predicting infections by multidrug-resistant bacteria.

Applications of Lab on a Chip in Antimicrobial Susceptibility of Staphylococcus aureus: A Systematic Review.

Background and Objectives: Staphylococcus aureus is a prevalent bacterium capable of inducing various infections, including skin and soft tissue infections, bloodstream infections, pneumonia, and surgical site infections. The emergence of antimicrobial resistance in S. aureus, particularly methicillin-resistant S. aureus, has raised substantial concerns within global healthcare settings. Prior to antibiotic prescription, the ideal approach is antimicrobial susceptibility testing (AST); however, this is frequently perceived as excessively complex and time-intensive. Lab-on-a-chip (LOC) technology holds promise in addressing these challenges and advancing fundamental microbiological research while also aiding in the development of therapeutic strategies. This systematic review aims to evaluate the potential utility of LOC for AST of S. aureus. Materials and Methods: This study adhered to the PRISMA guidelines. Various databases, including SCOPUS, PubMed/MEDLINE, SCIELO, and LILACS, in addition to gray literature sources, were employed in the review process. Results: Sixteen studies were included in this systematic review. All these studies detailed the effectiveness, rapidity, and predictability of LOC systems for assessing S. aureus susceptibility to various antibiotics. When comparing the LOC approach to traditional manual methods, it was evident that LOC requires a minimal quantity of reagents. Furthermore, most studies reported that the entire LOC procedure took 10 min to 7 h, with results being equally accurate as those obtained through traditional AST protocols. Conclusions: The potential application of LOC for AST of S. aureus is emphasized by its ability to provide rapid access to minimum inhibitory concentration data, which can substantially aid in selecting the most suitable antibiotics and dosages for treating challenging infections caused by this microorganism. Moreover, the rapid AST facilitated by LOC holds promise for enhancing the appropriateness and efficacy of therapy in clinical settings.

In vitro sensitivity of Malassezia furfur isolates from HIV-positive and negative patients to antifungal agents / Sensibilidad in vitro a antifúngicos de aislamientos de Malassezia furfur de pacientes positivos y negativos para HIV

INTRODUCTION: Malassezia is a lipophilic and lipid-dependent yeast genus belonging to the skin microbiota of humans and other animals. However, due to dysbiosis processes or other factors in the host, this yeast can cause different pathologies, ranging from skin diseases, such as seborrheic dermatitis, to fungemia. Isolation of Malassezia furfur has been reported in HIV-positive patients with or without skin lesions. Due to its opportunistic nature and its variable resistance to antifungal compounds, it is relevant to know the Malassezia sensitivity profiles. OBJECTIVE: To determine the sensitivity to different antifungal agents, of clinical isolates of M. furfur obtained from HIV-positive or negative patients, with or without seborrheic dermatitis. MATERIALS AND METHODS: Assessment of isolates sensitivity to itraconazole, voriconazole, fluconazole, and amphotericin B was performed by two techniques: (1) Broth microdilution using Clinical and Laboratory Standards Institute (CLSI) protocol M27-A3 with modifications; and (2) agar tests using Etest®. RESULTS: Isolates obtained from HIV patients showed an increase in the minimum inhibitory concentration of fluconazole, voriconazole, and amphotericin B, compared with those of non-HIV patients. Itraconazole was the antifungal with the lowest minimum inhibitory concentration (MIC) in most isolates. CONCLUSION: We observed differences in the sensitivity profiles of M. furfur isolates according to the context of the patient. High MIC of antifungals like fluconazole, commonly used for treating pathologies caused by Malassezia, were identified.

Introducción: Malassezia es un género de levaduras lipofílicas que dependen de los lípidos y hacen parte de la microbiota de la piel de humanos y otros animales. No obstante, debido a procesos de disbiosis u otros factores en el huésped, esta levadura puede llegar a causar diferentes enfermedades: desde cutáneas (como dermatitis seborreica) hasta fungemias. Se han reportado aislamientos de Malassezia furfur en pacientes positivos para HIV, con lesiones cutáneas o sin ellas. Por su carácter oportunista y sensibilidad variable a los compuestos antifúngicos, es relevante conocer los perfiles de sensibilidad. Objetivo: Determinar la sensibilidad a diferentes antifúngicos de aislamientos clínicos de M. furfur obtenidos de pacientes positivos o negativos para HIV, con dermatitis seborreica o sin ella. Materiales y métodos: La sensibilidad de los aislamientos a itraconazol, voriconazol, fluconazol y anfotericina B, se determinó mediante dos técnicas: microdilución en caldo según el protocolo M27-A3 del Clinical & Laboratory Standards Institute (CLSI), con modificaciones, y pruebas en agar mediante Etest®. Resultados: Los aislamientos obtenidos de pacientes con HIV mostraron aumento de la concentración inhibitoria mínima a fluconazol, voriconazol y anfotericina B, en comparación con los de pacientes sin HIV. Por otro lado, al evaluar la mayoría de los aislamientos, el itraconazol fue el antifúngico con la menor concentración inhibitoria mínima. Conclusión: Se evidencian diferencias en los perfiles de sensibilidad de los aislamientos de M. furfur, según el contexto del paciente, y elevadas concentraciones inhibitorias mínimas de antifúngicos como el fluconazol, usados comúnmente para el tratamiento de las enfermedades causadas por Malassezia spp.

Infecciones causadas por Staphylococcus aureus de la Comunidad. Estudio de sensibilidad y tendencias en población pediátrica. Años 2015 a 2020

Introducción: Staphylococcus aureus (SA) puede ocasionar cuadros infecciosos severos y muerte. La emergencia de cepas resistentes a meticilina constituye un desafío terapéutico. Objetivos: determinar el perfil de resistencia antimicrobiana de: Staphylococcus aureus adquirido en la comunidad (SA-CA), obtenidos de muestras biológicas de niños, entre 2015 a 2020. Material y Método: estudio descriptivo, observacional y retrospectivo. Las muestras para cultivos se extrajeron al ingreso hospitalario del paciente. Para determinación de resistencia y sensibilidad se utilizó normas de CSLI. Resultados: 244 aislamientos de SA-CA. Masculinos 99 (59%), menores de un año: 42 (25%), de 2 a 5 años: 34 (20%), de 6 a 11 años: 50 (30%) y entre 12 a 15 años: 42 (25%). De los aislados, 72% fueron SAMR (121/168) y 28% SAMS (47/168). Se observó un incremento de tasas anuales de aislamientos SAMR en infecciones de la comunidad desde el 2015 al 2020. Los aislamientos se originaron en piel y partes blandas 53,2 %; sangre 37,4%, orina 3,5%, LCR 2,4%, liquido articular 1,7%, abscesos profundos 1,2% y liquido pleural 0,6%. La prevalencia de SAMR-CA fue de 60,5 en el 2015, 59,6 %, 61,5%, 72,2 %, 67,3% y 75,5 % en los años sucesivos. No se aisló ninguna cepa resistente a la vancomicina. El 10,1% de las cepas estudiadas presentó resistencia inducida a la clindamicina. Conclusión: El SAMR se ha establecido como patógeno de la comunidad. La resistencia inducida por clindamicina fue del 10,1%. Un tercio de las infecciones fueron causadas por SAMS. Las prevalencias de SAMS muestran tendencia a la disminución.

Introduction: Staphylococcus aureus (SA) can cause severe infectious conditions and death. The emergence of methicillin-resistant strains constitutes a therapeutic challenge. Objectives: to determine the antimicrobial resistance profile of: Staphylococcus aureus acquired in the community (SA-CA), obtained from biological samples of children, between 2015 and 2020. Material and Method: descriptive, observational and retrospective study. The samples for cultures were extracted upon hospital admission of the patient. To determine resistance and sensitivity, CSLI standards were used. Results: 244 isolates of SA-CA. Males 99 (59%), under one-year-old: 42 (25%), from 2 to 5 years old: 34 (20%), from 6 to 11 years old: 50 (30%) and between 12 and 15 years old: 42 (25%). Of the isolates, 72% were SAMR (121/168) and 28% SAMS (47/168). An increase in annual rates of MRSA isolates in community infections was observed from 2015 to 2020. The isolates originated in skin and soft parts 53.2 %; blood 37.4%, urine 3.5%, CSF 2.4%, joint fluid 1.7%, deep abscesses 1.2% and pleural fluid 0.6%. The prevalence of MRSA-CA was 60.5 in 2015, 59.6%, 61.5%, 72.2%, 67.3%, and 75.5% in subsequent years. No vancomycin resistant strain was isolated. 10.1% of the strains studied presented induced resistance to clindamycin. Conclusion: MRSA has been established as a community pathogen. The resistance induced by clindamycin was 10.1%. One third of the infections was caused by SAMS. The prevalence of SAMS shows a downward trend.

Características epidemiológicas de la Osteomielitis aguda en niños internados en un hospital infeccioso de referencia

Introducción: La osteomielitis aguda es una infección del hueso que afecta principalmente a los niños y tiene generalmente diseminación hematógena, a veces asociada a un trauma. En la etiología influyen factores, como la edad, el estado inmunológico y las enfermedades concomitantes. En la mayoría de los casos, el principal agente etiológico es Staphylococcus aureus. Es importante el diagnóstico oportuno para evitar secuelas a mediano o largo plazo. Objetivo: Describir las características epidemiológicas de un grupo de pacientes con osteomielitis aguda. Métodos: Se realizó la revisión retrospectiva de los expedientes clínicos de pacientes egresados del servicio de pediatría del Instituto de Medicina Tropical, entre enero de 2016 y diciembre de 2020, con diagnóstico de osteomielitis aguda. Resultados: Los varones con osteomielitis corresponden al 67,8% del total de 59 casos registrados, en cuanto a los signos y síntomas, el dolor, la tumefacción y la impotencia funcional fueron predominantes, la fiebre se documentó en 49 (83,1%) pacientes, se registró antecedentes de cirugía en 37 (62,7%) de los pacientes y complicaciones en 42 (71,2%) de los pacientes, la complicación más frecuente fue osteomielitis crónica El sitio anatómico más frecuente fueron los miembros inferiores. El tratamiento empírico fue realizado con cefalosporinas de 3G en 72,9% de los pacientes, ya sea solo o combinado con clindamicina o vancomicina, un paciente con aislamiento de M. tuberculosis recibió tratamiento HRZE. Se aisló algún germen 44 pacientes (74,5%), el microorganismo predominante fue Staphylococcus aureus en 81,8 %, la mitad (52,3%) correspondieron a SAMR Se encontró una alta resistencia a oxacilina del 55,8% y un solo paciente resistente a clindamicina (2,2%). Conclusión Los hallazgos fueron similares a los reportados en la literatura en cuanto a etiología, sitio anatómico afectado y cobertura antibiótica.

Introduction: Acute osteomyelitis is a bone infection that mainly affects children and generally has hematogenous spread, sometimes associated with trauma. The etiology is influenced by factors such as age, immune status, and comorbidities. In most cases, the main etiologic agent is Staphylococcus aureus. Timely diagnosis is important to avoid sequelae in the medium or long term. Objective: To describe the epidemiological characteristics of a group of patients with acute osteomyelitis. Methods: A retrospective review of the clinical records of patients discharged from the pediatric service of the Institute of Tropical Medicine was carried out between January 2016 and December 2020, with a diagnosis of acute osteomyelitis. Results: Men with osteomyelitis correspond to 67.8% of the total of 59 registered cases, in terms of signs and symptoms, pain, swelling and functional impotence were predominant, fever was documented in 49 (83.1%) patients, a history of surgery was recorded in 37 (62.7%) of the patients and complications in 42 (71.2%) of the patients, the most frequent complication was chronic osteomyelitis The most frequent anatomical site was the lower limbs. Empirical treatment was performed with 3G cephalosporins in 72.9% of the patients, either alone or in combination with clindamycin or vancomycin. One patient with M. tuberculosis isolation received HRZE treatment. Some germ was isolated in 44 patients (74.5%), the predominant microorganism was Staphylococcus aureus in 81.8%, half (52.3%) corresponded to MRSA. A high resistance to oxacillin of 55.8% and a only patient resistant to clindamycin (2.2%). Conclusion The findings were similar to those reported in the literature in terms of etiology, affected anatomical site, and antibiotic coverage.

High levels of antimicrobial resistance in Escherichia coli and Salmonella from poultry in Ecuador.

Objective: To describe antimicrobial resistance profiles of Escherichia coli and Salmonella spp. isolated from chicken carcasses and the antimicrobials commonly used in animals in Ecuador and provide information on antimicrobial resistance patterns for implementing evidence-based corrective measures. Methods: Meat samples were collected from chicken carcasses in 199 slaughterhouses across Ecuador as part of a national pilot study for monitoring antimicrobial resistance in agricultural sources in 2019. Samples were tested for E. coli and Salmonella spp. Sensitivity to 10 critically important and three highly important antimicrobials (from a human health perspective) was assessed. The country report submitted to the World Organization for Animal Health was accessed to extract the quantity of antimicrobials produced or imported for use in animals. Results: Of 383 samples, E. coli was isolated from 148 (39%) and Salmonella spp. from 20 (5%) samples. Ninety percent of the isolates were resistant to at least one critically important antimicrobial. Resistance was highest to erythromycin (E. coli 76%; Salmonella spp. 85%) and tetracycline (E. coli 71%; Salmonella spp. 90%). Critically or highly important antimicrobials (colistin, tetracycline, trimethoprim/sulfamethoxazole) formed the bulk (87%) of antimicrobials used in animals as per the World Organization for Animal Health report. Conclusions: High prevalence of antimicrobial resistance in poultry in Ecuador calls for the development of guidelines and regulations on the use of antimicrobials and for engagement with livestock producers. The existing surveillance system needs to be strengthened to improve the monitoring of antimicrobial use and evolving resistance patterns.

Development of multiplex real-time PCR for detection of clarithromycin resistance genes for the Mycobacterium abscessus group.

Introduction. The M. abscessus molecular identification and its drug-resistance profile are important to choose the correct therapy.Aim. This work developed a multiplex real-time PCR (mqPCR) for detection of clarithromycin resistance genes for the Mycobacterium abscessus group.Methodology. Isolates received by Adolfo Lutz Institute from 2010 to 2012, identified by PCR restriction enzyme analysis of a fragment of the hsp65 gene (PRA-hsp65) as M. abscessus type 1 (n=135) and 2 (n=71) were used. Drug susceptibility test (DST) for CLA were performed with reading on days 3 and 14. Subespecies identification by hsp65 and rpoB genes sequencing and erm(41) and rrl genes for mutation detection and primer design were performed. erm(41) gene deletion was detected by conventional PCR. Primers and probes were designed for five detections: erm(41) gene full size and with deletion; erm(41) gene T28 and C28; rrl gene A2058.Results. In total, 191/206 (92.7 %) isolates were concordant by all methods and 13/206 (6.3 %) were concordant only between molecular methods. Two isolates (1.0 %) were discordant by mqPCR compared to rrl gene sequencing. The mqPCR obtained 204/206 (99.0 %) isolates in agreement with the gold standard, with sensitivity and specificity of 98 and 100 %, respectively, considering the gold standard method and 92 and 93 % regarding DST.Conclusion. The mqPCR developed by us proved to be an easy-to-apply tool, minimizing time, errors and contamination.

Molecular epidemiology and antifungal susceptibilities of Aspergillus species isolated from patients with invasive aspergillosis

SUMMARY OBJECTIVE: The aim of this study was to evaluate the demographic data, molecular epidemiology, and in vitro antifungal susceptibility results of patients with Aspergillus isolated from various clinical specimens. METHODS: A total of 44 Aspergillus strains were studied. The definition of invasive aspergillosis in patients was made according to European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) criteria. Strains were phenotypically and molecularly identified. Demographic characteristics of patients and genotypes of strains were evaluated. Phylogenetic analysis was done by the The Unweighted Pair-Group Method with Arithmetic Mean (UPGMA). Antifungal susceptibility of strains was determined according to The Clinical and Laboratory Standards Institute (CLSI)-M61-Ed2 and The European Committee on Antimicrobial Susceptibility Testing (EUCAST). RESULTS: A total of 11 patients were classified as proven and 33 as probable invasive aspergillosis. There was a statistically significant difference in age groups, subdisease, neutropenic, and receiving chemotherapy between groups. A total of 23 strains were identified as Aspergillus fumigatus, 12 as Aspergillus niger, 6 as Aspergillus flavus, and 3 as Aspergillus terreus. Phylogenetic analysis revealed five different genotypes. No statistical difference was found in the comparisons between patients groups and genotype groups. There was a statistically significant difference between genotype groups and voriconazole, posaconazole, and itraconazole Minimum Inhibition Concentration (MIC). CONCLUSION: Accurate identification of strains and antifungal susceptibility studies should be performed due to azole and amphotericin B resistance. Genotyping studies are important in infection control due to identifying sources of infection and transmission routes.

In vitro sensitivity of Malassezia furfur isolates from HIV-positive and negative patients to antifungal agents / Sensibilidad in vitro a antifúngicos de aislamientos de Malassezia furfur de pacientes positivos y negativos para HIV

Introduction. Malassezia is a lipophilic and lipid-dependent yeast genus belonging to the skin microbiota of humans and other animals. However, due to dysbiosis processes or other factors in the host, this yeast can cause different pathologies, ranging from skin diseases, such as seborrheic dermatitis, to fungemia. Isolation of Malassezia furfur has been reported in HIV-positive patients with or without skin lesions. Due to its opportunistic nature and its variable resistance to antifungal compounds, it is relevant to know the Malassezia sensitivity profiles. Objective. To determine the sensitivity to different antifungal agents, of clinical isolates of M. furfur obtained from HIV-positive or negative patients, with or without seborrheic dermatitis. Materials and methods. Assessment of isolates sensitivity to itraconazole, voriconazole, fluconazole, and amphotericin B was performed by two techniques: (1) Broth microdilution using Clinical and Laboratory Standards Institute (CLSI) protocol M27-A3 with modifications; and (2) agar tests using Etest®. Results. Isolates obtained from HIV patients showed an increase in the minimum inhibitory concentration of fluconazole, voriconazole, and amphotericin B, compared with those of non-HIV patients. Itraconazole was the antifungal with the lowest minimum inhibitory concentration (MIC) in most isolates. Conclusion. We observed differences in the sensitivity profiles of M. furfur isolates according to the context of the patient. High MIC of antifungals like fluconazole, commonly used for treating pathologies caused by Malassezia, were identified.

Introducción. Malassezia es un género de levaduras lipofílicas que dependen de los lípidos y hacen parte de la microbiota de la piel de humanos y otros animales. No obstante, debido a procesos de disbiosis u otros factores en el huésped, esta levadura puede llegar a causar diferentes enfermedades: desde cutáneas (como dermatitis seborreica) hasta fungemias. Se han reportado aislamientos de Malassezia furfur en pacientes positivos para HIV, con lesiones cutáneas o sin ellas. Por su carácter oportunista y sensibilidad variable a los compuestos antifúngicos, es relevante conocer los perfiles de sensibilidad. Objetivo. Determinar la sensibilidad a diferentes antifúngicos de aislamientos clínicos de M. furfur obtenidos de pacientes positivos o negativos para HIV, con dermatitis seborreica o sin ella. Materiales y métodos. La sensibilidad de los aislamientos a itraconazol, voriconazol, fluconazol y anfotericina B, se determinó mediante dos técnicas: microdilución en caldo según el protocolo M27-A3 del Clinical & Laboratory Standards Institute (CLSI), con modificaciones, y pruebas en agar mediante Etest®. Resultados. Los aislamientos obtenidos de pacientes con HIV mostraron aumento de la concentración inhibitoria mínima a fluconazol, voriconazol y anfotericina B, en comparación con los de pacientes sin HIV. Por otro lado, al evaluar la mayoría de los aislamientos, el itraconazol fue el antifúngico con la menor concentración inhibitoria mínima. Conclusión. Se evidencian diferencias en los perfiles de sensibilidad de los aislamientos de M. furfur, según el contexto del paciente, y elevadas concentraciones inhibitorias mínimas de antifúngicos como el fluconazol, usados comúnmente para el tratamiento de las enfermedades causadas por Malassezia spp.

High levels of antimicrobial resistance in Escherichia coli and Salmonella from poultry in Ecuador / Niveles elevados de resistencia antimicrobiana de las bacterias Escherichia coli y Salmonella en aves de corral en Ecuador / Altos níveis de resistência aos antimicrobianos em Escherichia coli e Salmonella de aves de criação no Equador

ABSTRACT Objective. To describe antimicrobial resistance profiles of Escherichia coli and Salmonella spp. isolated from chicken carcasses and the antimicrobials commonly used in animals in Ecuador and provide information on antimicrobial resistance patterns for implementing evidence-based corrective measures. Methods. Meat samples were collected from chicken carcasses in 199 slaughterhouses across Ecuador as part of a national pilot study for monitoring antimicrobial resistance in agricultural sources in 2019. Samples were tested for E. coli and Salmonella spp. Sensitivity to 10 critically important and three highly important antimicrobials (from a human health perspective) was assessed. The country report submitted to the World Organization for Animal Health was accessed to extract the quantity of antimicrobials produced or imported for use in animals. Results. Of 383 samples, E. coli was isolated from 148 (39%) and Salmonella spp. from 20 (5%) samples. Ninety percent of the isolates were resistant to at least one critically important antimicrobial. Resistance was highest to erythromycin (E. coli 76%; Salmonella spp. 85%) and tetracycline (E. coli 71%; Salmonella spp. 90%). Critically or highly important antimicrobials (colistin, tetracycline, trimethoprim/sulfamethoxazole) formed the bulk (87%) of antimicrobials used in animals as per the World Organization for Animal Health report. Conclusions. High prevalence of antimicrobial resistance in poultry in Ecuador calls for the development of guidelines and regulations on the use of antimicrobials and for engagement with livestock producers. The existing surveillance system needs to be strengthened to improve the monitoring of antimicrobial use and evolving resistance patterns.

RESUMEN Objetivo. Describir los perfiles de resistencia antimicrobiana de las bacterias Escherichia coli y Salmonella spp. aisladas en carne de pollo y los antimicrobianos comúnmente empleados en animales en Ecuador, así como proporcionar información sobre los patrones de resistencia a los antimicrobianos para poner en marcha medidas correctivas basadas en la evidencia. Métodos. Se recogieron muestras de carne de pollo en 199 mataderos de todo Ecuador en el marco de un estudio piloto nacional para monitorear la resistencia a los antimicrobianos en fuentes agrícolas en el 2019. Se analizaron las muestras en busca de E. coli y Salmonella spp. Se evaluó la sensibilidad a diez antimicrobianos de importancia crítica y tres muy importantes (para la salud humana). Se accedió al informe de país presentado ante la Organización Mundial de Sanidad Animal para obtener la cantidad de antimicrobianos producidos o importados para su uso en animales. Resultados. De 383 muestras, se aisló E. coli en 148 (39%) y Salmonella spp. en 20 (5%). En total, 90% de las cepas aisladas fueron resistentes a al menos un antimicrobiano de importancia crítica. Hubo una mayor resistencia a la eritromicina (E. coli: 76%; Salmonella spp.: 85%) y a la tetraciclina (E. coli: 71%; Salmonella spp.: 90%). Los antimicrobianos de importancia crítica o muy importantes (colistina, tetraciclina, trimetoprima/sulfametoxazol) constituyeron la mayor parte (87%) de los antimicrobianos empleados en animales según el informe de la Organización Mundial de Sanidad Animal. Conclusiones. Debido a la alta prevalencia de la resistencia a los antimicrobianos en las aves de corral en Ecuador, son imprescindibles la elaboración de directrices y regulaciones sobre el uso de antimicrobianos y el compromiso con los productores pecuarios. Es necesario fortalecer el sistema de vigilancia existente para mejorar el seguimiento del uso de antimicrobianos y de la evolución de los patrones de resistencia.

RESUMO Objetivo. Descrever perfis de resistência aos antimicrobianos em Escherichia coli e Salmonella spp. isoladas de carcaças de frango e os antimicrobianos comumente usados em animais no Equador e fornecer informações sobre padrões de resistência aos antimicrobianos para implementar medidas corretivas baseadas em evidências. Métodos. Foram coletadas amostras de carne de carcaças de frango em 199 abatedouros em todo o Equador como parte de um estudo piloto nacional para monitorar a resistência aos antimicrobianos de origem agrícola em 2019. Foram testadas amostras de E. coli e Salmonella spp. Foi avaliada a sensibilidade a 10 agentes antimicrobianos de importância crítica e três agentes antimicrobianos muito importantes (do ponto de vista da saúde humana). O relatório do país apresentado à Organização Mundial de Saúde Animal foi acessado para extrair a quantidade de antimicrobianos produzidos ou importados para uso em animais. Resultados. De 383 amostras, E. coli foi isolada em 148 (39%) e Salmonella spp. em 20 (5%). Noventa por cento dos isolados foram resistentes a pelo menos um antimicrobiano de importância crítica. A resistência foi maior à eritromicina (E. coli, 76%; Salmonella spp., 85%) e à tetraciclina (E. coli, 71%; Salmonella spp., 90%). Antimicrobianos de importância crítica ou muito importantes (colistina, tetraciclina, trimetoprim/sulfametoxazol) responderam pela maior parte (87%) dos antimicrobianos utilizados em animais, conforme o relatório da Organização Mundial de Saúde Animal. Conclusões. A alta prevalência de resistência aos antimicrobianos na avicultura no Equador exige o desenvolvimento de diretrizes e regulamentos sobre o uso de antimicrobianos e o envolvimento com os produtores de gado e avícolas. O sistema de vigilância existente precisa ser reforçado para melhorar o monitoramento do uso de antimicrobianos e a evolução dos padrões de resistência.

Update on Pediatric Sepsis in Mexico.

The main objective of this work was to determine and update the causal agents' antibiotic sensitivity and resistance patterns on pediatric sepsis in a population of northeast Mexico. It is a cross-sectional study showing the results of blood cultures of pediatric patients with a presumptive diagnosis of sepsis were reviewed according to the SOFA criteria during 2020 in a public hospital in Mexico. A total of 207 blood cultures were performed and analyzed. The main isolated microorganisms were Staphylococcus, followed by Klebsiella and Escherichia. Several microorganisms showed 100% of sensitivity to different antibiotics or antifungals, some of them include Vancomycin, Voriconazole, Meropenem, Ciprofloxacin, and Cefotaxime. Bacteria of genre Staphylococcus showed its highest sensitivity rate to Tigecycline with 63.3%. Too Staphylococcus showed the highest resistance rate to Oxacillin with 50%. Although the patterns of sepsis-causing germs are similar to those previously reported, the development of new drugs with greater efficacy is the main contribution.

Susceptibilidad antimicrobiana de Streptococcus agalactiae en un hospital materno-infantil de Lima, Perú. 2015-2020

Objetivo : Determinar la susceptibilidad antimicrobiana de cepas de Streptococcus agalactiae (EGB) aisladas entre 2015-2020 en un hospital materno-infantil. Material y métodos : Estudio descriptivo realizado entre 2015 y 2020. La identificación y susceptibilidad antimicrobiana se determinaron por el sistema automatizado Vitek® 2. La base de datos obtenida fue transferida al programa WHONET versión 5.6. Se incluyeron 506 aislamientos de EGB. Resultados : 500 (98,8%) fueron en mujeres. La edad media fue de 31,9 ± 11,8 años. Todos fueron sensibles a penicilina, ampicilina y vancomicina, la resistencia a eritromicina, clindamicina y levofloxacino fue de 38,5%, 37,4% y 28%, respectivamente, mostrando un pico de resistencia en 2018. Conclusión : Se observó un incremento en la resistencia de EGB a los antimicrobianos, especialmente el año 2018.

SUMMARY Objective : To determine the antimicrobial susceptibility of S. agalactiae (SA) strains isolated from 2015-2020 in a maternal-infant hospital in Lima, Peru. Methods : A descriptive study was carried-out from 2015 to 2020. Identification and susceptibility were performed using Vitek®2. Data base was transferred to the WHONET version 5.6 program; 506 isolates were analyzed. Results: 500 (98.8%) were females; mean age was 31.9 ± 11.8 years. All strains were susceptible to penicillin, ampicillin and vancomycin. Resistance to erythromycin, clindamycin and levofloxacin were 38.5%, 37.4% and 28%, respectively, showing the highest value in 2018. Conclusions : An increase in resistance of SA to antimicrobials was observed, especially in 2018.

Sensitivity of different DNA extraction methods and PCR to detect resistance in patients with leprosy stratified by the bacilloscopic index.

INTRODUCTION: Antimicrobial resistance in leprosy is an emerging problem, and the quantitative impact of low bacilloscopic indexes (BIs) on the sensitivity of molecular tests is unknown. We aimed to evaluate the sensitivity of gene sequencing for the detection of mutations related to antimicrobial resistance in Mycobacterium leprae in patients with low BIs using an analytical model. METHODS: Patients with leprosy were included and divided into two groups depending on their BIs (≥ 2+ and < 2+). The sensitivities of the two DNA extraction methods were compared after amplifying and sequencing the repetitive element (RLEP), folP1, rpoB and gyrA in M. leprae. RESULTS: We included 56 patients with leprosy: 35 had BIs less than 2+ (22 had negative slit-skin smear [SSS] results) and 21 patients had BIs greater than or equal to 2+. The sensitivity of the amplification of the RLEP target and the gene sequencing of folP1, rpoB and gyrA was 50 to 70% lower in patients with a BI less than 2+ and was significantly reduced in patients with lower BIs for all targets (p < 0.001). One patient had a mutation in the folP1 gene, and 14 patients had mutations in the gyrA gene, but no mutations related to antimicrobial resistance were found. CONCLUSIONS: We can conclude that the sensitivity of molecular tests is directly related to the BI, but these tests can still detect up to 20% of the targets in patients with BIs < 2+. New strategies to improve the sensitivity for detecting antimicrobial resistance in leprosy patients and reasonable clinical criteria for follow-up and the introduction of alternative treatments must be developed.

Evaluation of phenotypic methods for detection of polymyxin B-resistant bacteria.

Determination of sensitivity to polymyxins has always been a challenge, especially in clinical laboratory routines. This study evaluated two rapid, simple, and inexpensive phenotypic methods to test polymyxin B (PMB) susceptibility in Enterobacterales and non-fermenting Gram-negative bacilli. One hundred isolates were used in the tests. The isolates were collected in three hospitals in southern and southeastern Brazil from 1995 to 2019. We compared broth microdilution (reference method) with the broth disk elution test and modified drop test, using polymyxin B -disk or PMB -powder in 2 concentrations (12 and 16 µg/ml). For the broth disk elution and modified drop test with the concentration of 12 µg/ml, categorical agreement values exceeded 90%. The modified drop test with a concentration of 12 µg/ml and broth disk elution may be excellent for initial screening of polymyxin-resistance in laboratory routines. Moreover, these methods are simple and use inexpensive supplies, and may optimize therapeutic decisions.

Evaluación de la Susceptibilidad Antimicrobiana de Porphyromonas gingivalis Aisladas de Pacientes Periodontales en Población Chilena

RESUMEN: Porphyromonas gingivalis (P. gingivalis) es un microorganismo frecuentemente aislado en pacientes con periodontitis, en los que también se incluye los de la población chilena. El modelo de "Keystone bacteria" demostró que P. gingivalis induce la disbiosis del biofilm subgingival y permite el desarrollo de algunas especies sobre otras, modulando la patogenicidad de la comunidad microbiológica completa. El tratamiento de la periodontitis es principalmente mecánico, pero en condiciones específicas es necesario el complemento con antibioterapia. Los estudios globales de antibióticos evaluados en ensayos clínicos y estudios in vitro han mostrado resultados mixtos en cuanto a eficacia y susceptibilidad. Este estudio descriptivo tuvo como objetivo evaluar el perfil de susceptibilidad antimicrobiana in vitro a metronidazol, clindamicina, amoxicilina más ácido clavulánico, moxifloxacino y azitromicina de P. gingivalis aisladas de pacientes periodontales chilenos. Se obtuvieron muestras microbiológicas de pacientes con diagnóstico de periodontitis estadíos III y IV generalizada, las que se sometieron a procesos de identificación mediante un espectrómetro de masas (MALDI-TOF MS). Posteriormente, a cada muestra positiva a P. gingivalis se aplicó el protocolo gold standard de susceptibilidad para los cinco antimicrobianos evaluados (Dilución en Agar Sangre Brucella- McFarland 0.5). Se seleccionaron 50 pacientes (25 mujeres, 25 hombres) entre 34-69 años. Finalmente, se recuperaron 25 cepas de P. gingivalis (50 %) para el análisis de susceptibilidad y todas ellas fueron sensibles a todos los antibióticos evaluados (100 % susceptibilidad). Las cepas de P. gingivalis fueron altamente sensibles a los cinco antibióticos evaluados en esta población, lo que podría implicar contar con diferentes alternativas de tratamiento farmacológico antimicrobi ano como complemento al tratamiento mecánico convencional en pacientes específicos.

ABSTRACT: Porphyromonas gingivalis (P. gingivalis) is a microorganism frequently isolated in patients with periodontitis, which also include those of Chilean population. The "Keystone bacteria" model demonstrated that P. gingivalis induces dysbiosis of the subgingival biofilm and allows the development of some species over others, modulating the pathogenicity of the entire microbiological community. The treatment of periodontitis is mainly mechanical; nevertheless, under specific conditions the complement with antibiotherapy is needed. Global studies of antibiotics evaluated in clinical trials and in vitro studies have shown mixed results in terms of efficacy and susceptibility. This descriptive study aimed to evaluate antimicrobial susceptibility profile in vitro to metronidazole, clindamycin, amoxicillin plus clavulanic acid, moxifloxacin and azithromycin of P. gingivalis isolated from Chilean periodontal patients. Microbiological samples were obtained from patients with a diagnosis of generalized periodontitis stages III and IV, which were exposed to identification processes by a mass spectrometer (MALDI-TOF MS). Subsequently, the gold standard susceptibility protocol for the five antimicrobials evaluated was applied to each P. gingivalis- positive sample (Dilution in Brucella-McFarland Blood Agar 0.5). 50 patients (25 women, 25 men) between 34-69 years old were selected. Finally, 25 P. gingivalis strains (50 %) were recovered for susceptibility testing and all of them were susceptible to all antibiotics tested (100 % susceptibility). P. gingivalis strains were highly susceptible to the five antibiotics evaluated in this population, which could implies counting different antimicrobial pharmacological treatment alternatives as a complement to conventional mechanical treatment in specific patients.