Crotamine derived from Crotalus durissus terrificus venom combined with drugs increases in vitro antibacterial and antifungal activities.

This study investigated crotamine (CTA), a peptide derived from the venom of the South American rattlesnake Crotalus durissus terrificus, known for its exceptional cell penetration potential. The objective was to explore the antibacterial and antifungal activity of CTA, its ability to inhibit efflux pumps and evaluate the effectiveness of its pharmacological combination with antibiotics and antifungals. In microbiological assays, CTA in combination with antibiotics was tested against strains of S. aureus and the inhibition of NorA, Tet(K) and MepA efflux pumps was also evaluated. CTA alone did not present clinically relevant direct antibacterial action, presenting MIC > 209.7 µM against strains S. aureus 1199B, IS-58, K2068. The standard efflux pump inhibitor CCCP showed significant effects in all negative relationships to assay reproducibility. Against the S. aureus 1199B strain, CTA (20.5 µM) associated with norfloxacin diluted 10 × (320.67 µM) showed a potentiating effect, in relation to the control. Against the S. aureus IS-58 strain, the CTA associated with tetracycline did not show a significant combinatorial effect, either with 2304 or 230.4 µM tetracycline. CTA at a concentration of 2.05 µM associated with ciprofloxacin at a concentration of 309.4 µM showed a significant potentiating effect. In association with EtBr, CTA at concentrations of 2.05 and 20.5 µM potentiated the effect in all strains tested, reducing the prevention of NorA, Tet(K) and MepA efflux pumps. In the C. albicans strain, a potentiating effect of fluconazole (334.3 µM) was observed when combined with CTA (2.05 µM). Against the C. tropicalis strain, a significant effect was also observed in the association of fluconazole 334.3 µM, where CTA 2.05 µM considerably reduced fungal growth and decreased the potentiation of fluconazole. Against the C. krusei strain, no significant potentiating effect of fluconazole was obtained by CTA. Our results indicate that CTA in pharmacological combination potentiates the effects of antibiotics and antifungal. This represents a new and promising antimicrobial strategy for treating a wide variety of infections.

Concerns about the use of antimicrobials in swine herds and alternative trends.

Pig productivity in Brazil has advanced a lot in recent decades. Specialized breeds are more vulnerable to pathogens, which has boosted the use of antimicrobials by farmers. The selective pressure generated favors the emergence of resistant bacteria, which compromises the effectiveness of this treatment and limits therapeutic options. In addition to increasing costs and mortality rates in the production system, public awareness of this issue has increased. The authorities have imposed restrictive measures to control the use of antimicrobials and have banned their use as growth promoters. This literature review highlights biosecurity and animal welfare to prevent pig diseases. Hence, we describe alternatives to the use of antimicrobials in pig production for the selection of effective non-antibiotic feed additives that help maintain good health and help the pig resist disease when infection occurs.

Combining eugenol and dihydroeugenol with a piperazine moiety to create new antimicrobial agents that are effective against resistant species.

Historically, the piperazine moiety has been demonstrated to possess pharmacophoric properties, and has subsequently been incorporated in many drugs that have antitumor, antimalarial, antiviral, antibacterial and antifungal properties. Derivatives of eugenol and dihydroeugenol have also been reported as being bioactive compounds. This study reports the synthesis of a range of eugenol/dihydroeugenol - piperazine derivatives which have been tested as antimicrobial compounds against Gram positive, Gram negative and rapid-growing mycobacteria (RGM). The rationale employed in the design of the structural pattern of these new derivatives, provides useful insights into the structure-activity relationships (SAR) of the series. Antimicrobial activity tests were extremely encouraging, with the majority of the synthesised compounds being more active than eugenol and dihydroeugenol starting materials. The antimicrobial potential was most notable against the Gram-negative species K. pneumoniae and P. aeruginosa, but there was also significant performance against the Gram-positive strains S. epidermidis and S. aureus and the Rapidly Growing Mycobacteria (RGM) strains tested. Tests using the synthesised compounds against multidrug-resistance clinical (MDR) isolates also showed high activity. The biofilm inhibition tests using M. fortuitum showed that all evaluated derivatives were able to inhibit biofilm formation even at low concentrations. In terms of structural-activity relationships; the results generated by this study demonstrate that the compounds with bulky substituents on the piperazine subunit were much more active than those with less bulky groups, or no groups. Importantly, the derivatives with a sulfonamide side chain were the most potent compounds. A further observation was that those compounds with a para-substituted benzenesulfonamide ring stand out, regardless of whether this substituent is a donor or an electron-withdrawing group.

Infections in Children with Cancer Admitted in an Oncology Reference Hospital: A Cross-sectional Study.

Pediatric oncology patients are usually immunosuppressed due to factors such as the neoplasm and its treatment, making them more susceptible to infections. This article aims to determine the infection profile of pediatric oncology patients admitted to an oncology reference hospital in Natal, Rio Grande do Norte, Brazil. A retrospective and cross-sectional study was conducted, collecting data from patients hospitalized due to infection in a pediatric oncology unit exclusively for the Brazilian public health system, spanning from 2018 to 2021. A total of 168 episodes of infections were identified in 96 patients, resulting in 157 hospitalizations. Among the patients with infections, 62.4% had hematological malignancies, and out of these cases, 74.6% specifically had Acute Lymphoid Leukemia. The Escherichia coli (31.9%) was the most prevalent microorganism isolated from the samples. Multidrug-resistant microorganisms accounted for 52% of all identified microorganisms. Fluoroquinolones and beta-lactam were the most prevalent antibiotic classes in the analyzed antibiograms. Factors such as Sex, type of cancer, chemotherapy in the last 30 days, were found to be associated with the occurrence of infection (p < 0.05). Conducting epidemiological studies regarding infections in pediatric oncology is crucial to development of empirical protocols, and the implementation of strategies to better control future infections.

Challenges and advances in antimicrobial peptide development.

Microbial resistance is a major concern for public health worldwide, mainly because of the inappropriate use of antimicrobials. In this scenario, antimicrobial peptides (AMPs) have emerged as a potential therapeutic alternative means by which to control infectious diseases, because of their broad spectrum of action. However, some challenges can make their clinical application problematic, including metabolic instability and toxicity. Here, we provide a clear description of AMPs as promising molecules for the development of unusual antimicrobial drugs. We also describe current strategies used to overcome the main difficulties related to AMP clinical application, including different peptide designs and nanoformulation.

Untargeted Metabolomics for Unraveling the Metabolic Changes in Planktonic and Sessile Cells of Salmonella Enteritidis ATCC 13076 after Treatment with Lippia origanoides Essential Oil.

Nontyphoidal Salmonella species are one of the main bacterial causes of foodborne diseases, causing a public health problem. In addition, the ability to form biofilms, multiresistance to traditional drugs, and the absence of effective therapies against these microorganisms are some of the principal reasons for the increase in bacterial diseases. In this study, the anti-biofilm activity of twenty essential oils (EOs) on Salmonella enterica serovar Enteritidis ATCC 13076 was evaluated, as well as the metabolic changes caused by Lippia origanoides thymol chemotype EO (LOT-II) on planktonic and sessile cells. The anti-biofilm effect was evaluated by the crystal violet staining method, and cell viability was evaluated through the XTT method. The effect of EOs was observed by scanning electron microscopy (SEM) analysis. Untargeted metabolomics analyses were conducted to determine the effect of LOT-II EO on the cellular metabolome. LOT-II EO inhibited S. Enteritidis biofilm formation by more than 60%, without decreasing metabolic activity. Metabolic profile analysis identified changes in the modulation of metabolites in planktonic and sessile cells after LOT-II EO treatment. These changes showed alterations in different metabolic pathways, mainly in central carbon metabolism and nucleotide and amino acid metabolism. Finally, the possible mechanism of action of L. origanoides EO is proposed based on a metabolomics approach. Further studies are required to advance at the molecular level on the cellular targets affected by EOs, which are promising natural products for developing new therapeutic agents against Salmonella sp. strains.

Photodynamic inactivation of different Candida species and inhibition of biofilm formation induced by water-soluble porphyrins.

BACKGROUND: Candida spp. is the main fungal genus related to infections in humans, and its treatment has become a challenge due to the production of biofilm and its resistance/multi-resistance profile to conventional antifungals. Antimicrobial photodynamic therapy stands out as a treatment characterized by a broad spectrum of antimicrobial action, being able to induce oxidative stress in pathogens, and porphyrins are photosensitizers with high selectivity to pathogens. Thus, this work aimed to analyze the photoinactivation of different species of Candida by two cationic (4-H2TMeP+ and 3-H2TMeP+) and one anionic (4-H2TPSP‒) porphyrins. MATERIALS AND METHODS: Microdilution assays were performed to determine the MIC100, with subsequent determination of MFC100. Determination of oxidative species was done through the use of scavengers, while biofilm morphological features were investigated using the atomic force microscopy. RESULTS: Cationic porphyrins were significantly efficient in inactivating Candida albicans and non-albicans species with 100% growth inhibition and fungicidal activity (MFC100/MIC100 ≤ 4.0). The cationic porphyrins were also able to interfere in Candida spp biofilm formation. The photo-oxidative mechanism activated by 3-H2TMeP+ in Candida spp. is concurrent with the production of singlet oxygen and oxygen radical species. In the AFM analysis, 3-H2TMeP+ was able to reduce yeast adhesion to the surface. CONCLUSIONS: Cationic porphyrins can photo-inactivate different species of Candida in both planktonic and biofilm-associated forms, and reduce the adhesion of these fungi to the surface.

Knowledge, attitudes, and behavior regarding antibiotics, antibiotic use, and antibiotic resistance in students and health care professionals of the district of Barranquilla (Colombia): A cross-sectional survey.

Objective: To evaluate the knowledge, attitudes and behavior regarding antibiotics, use of antibiotics, and antibiotic resistance in students and health care professionals of the district of Barranquilla, Colombia. Study design: Descriptive, cross-sectional. Methods: A sample of 399 respondents was selected, that included health professionals and medical students from 12 health institutions in the district of Barranquilla (Colombia), using an established stratified sampling method. Each of the respondent professionals completed a survey that included 43 items in the Likert scale. A descriptive analysis of the study variables was performed using the software SPSS version 25. Results: Most of the respondents were women (64.4%), aged between 26 and 35 years (47.6%); 28.8% were nurses and 26.1% general practitioners, with ≤10 years of professional experience (63.4%). Overall, the survey revealed that the participants had considerable knowledge about antibiotic use (89.5%-98% correct answers) and the spread of antibiotic resistance (67.4%-89% correct answers). Approximately 74% of the respondents agreed or fully agreed with the questions related to the management of infections and the provision of advice. Conclusions: The present study revealed that most of the health care professionals surveyed had a good knowledge about antibiotic use, although strategies must be developed to strengthen knowledge regarding the spread of antibiotic resistance. Likewise, it is important to identify opportunities for improvement related with access to the guidelines and/or materials necessary to treat infections and to provide advice on antibiotic use and antibiotic resistance.

Plant Lectins: A Review on their Biotechnological Potential Toward Human Pathogens.

The indiscriminate use of antibiotics is associated with the appearance of bacterial resistance. In light of this, plant-based products treating infections are considered potential alternatives. Lectins are a group of proteins widely distributed in nature, capable of reversibly binding carbohydrates. Lectins can bind to the surface of pathogens and cause damage to their structure, thus preventing host infection. The antimicrobial activity of plant lectins results from their interaction with carbohydrates present in the bacterial cell wall and fungal membrane. The data about lectins as modulating agents of antibiotic activity, potentiates the effect of antibiotics without triggering microbial resistance. In addition, lectins play an essential role in the defense against fungi, reducing their infectivity and pathogenicity. Little is known about the antiviral activity of plant lectins. However, their effectiveness against retroviruses and parainfluenza is reported in the literature. Some authors still consider mannose/ glucose/N-Acetylglucosamine binding lectins as potent antiviral agents against coronavirus, suggesting that these lectins may have inhibitory activity against SARS-CoV-2. Thus, it was found that plant lectins are an alternative for producing new antimicrobial drugs, but further studies still need to decipher some mechanisms of action.

Prevenção e controle de resistência aos antimicrobianos na Atenção Primária à Saúde: evidências para políticas / Prevention and control of antimicrobial resistance in Primary Health Care: evidence for policies

Resumo A resistência aos antimicrobianos é um problema mundial que põe em risco a segurança da saúde da população. O objetivo deste artigo é identificar e avaliar estratégias para prevenção e controle de resistência microbiana, bem como barreiras para sua implementação em serviços de Atenção Primária à Saúde (APS). Realizou-se uma síntese de evidências para políticas. As buscas de evidências foram realizadas entre novembro/dezembro de 2018, em 13 bases de dados. Um diálogo deliberativo foi realizado para validação dos resultados e levantamento de barreiras e facilitadores para implementação das estratégias. As 13 revisões sistemáticas incluídas mostraram que intervenções com foco em educação, uso de sistemas eletrônicos e biomarcadores reduziram o consumo e prescrição de antimicrobianos. É um obstáculo à implementação a expectativa de usuários/cuidadores em receber prescrição de antibióticos, e são facilitadores as ações educativas que envolvem profissionais de saúde. O uso racional de medicamentos se impõe na APS com vistas à prevenção da resistência dos microrganismos aos antibióticos. As intervenções identificadas neste estudo podem ser implementadas isoladamente ou em conjunto, conforme o contexto local.

Abstract Antimicrobial resistance is a global problem that puts the population's health at risk. This paper aims to identify and evaluate strategies for the prevention and control of antimicrobial resistance, and barriers to their implementation in Primary Health Care (PHC) services. We developed an evidence brief for policies. The search for evidence occurred in 13 databases from November to December 2018. A deliberative dialogue was performed to validate the results and we identified barriers and facilitators to implementing the strategies. The 13 systematic reviews included evidenced that the interventions focused on education, use of electronic systems and biomarkers reduced antimicrobial consumption and prescription. User/caregiver's expectation to receive antibiotic prescriptions was the main obstacle to implementing strategies, while education actions involving health professionals were facilitators. The rational use of medications in the PHC services is crucial to prevent antimicrobial resistance to antibiotics. The interventions identified in this study can be implemented alone or combined, according to local context.

Prevalence of antimicrobial-resistant Enterococcus faecium in commercial cattle: a systematic review and meta-analysis / Prevalência de Enterococcus faecium resistente a antimicrobianos na bovinocultura comercial: uma revisão sistemática e meta-análise

The use of antimicrobials in animals is broader compared to humans, which can influence the increase in microbial resistance. This study was a systematic review which determined the prevalence of resistant Enterococcus faecium in commercial cattle. Eighteen studies were included, mainly carried out in European countries (n=9) and in the production (n= 11) and retail (n= 7) environments. The main material used in the detection of the microorganism was milk. The mean prevalence of resistant E. faecium in cattle was 4.3% (95% CI = 2.8-5.0%), but the prevalence in Asia was higher [25.4% (95% CI = 20.5-30.6%)]. There was a higher prevalence in samples from retail (13.7%; 95% CI=11.5-16.1%) and collected mainly from equipment surfaces (12.5%; 95% CI= 5.5-26.1%) than in the others tested samples. Antibiotics frequently tested were vancomycin, tetracycline, ciprofloxacin, and erythromycin, with resistance percentages of 50%, 59%, 79%, and 94%, respectively. These results reinforce the need to plan interventions to reduce antimicrobials in food-producing animals.

O uso de antimicrobianos em animais é mais frequente quando comparado aos humanos, e isso pode influenciar no desenvolvimento da resistência microbiana. O presente estudo teve como objetivo realizar uma revisão sistemática cujo desfecho de interesse foi a prevalência de E. faecium resistente a antimicrobianos na bovinocultura comercial. Foram incluídos 18 estudos, realizados principalmente em países europeus (n=9), em ambientes de produção (n=11) e destinados ao varejo (n=7). O principal material utilizado na detecção do microrganismo foi o leite. A prevalência de E. faecium resistente em bovinos foi de 4,3% (IC 95% -2,8-5,0%), mas a prevalência na Ásia foi maior [25,4% (IC 95%=20,5-30,6%)]. Houve maior prevalência em amostras do varejo (13,7%; IC 95%=11,5-16,1%) e coletadas principalmente de superfícies de equipamentos (12,5%; IC 95%-5,5-26,1%). Os antibióticos frequentemente testados foram vancomicina, tetraciclina, ciprofloxacino, e eritromicina, com percentuais de resistência de 50%, 59%, 79%, e 94%, respectivamente. Estes resultados reforçam a necessidade de intervenções planejadas para reduzir a utilização de antimicrobianos nos animais criados para produção de alimentos.

Escherichia coli productora de BLEE de origen comunitario e intrahospitalario / BLEE-producing Escherichia coli of community and hospital-acquired origin / Escherichia coli produtora de BLEE de origem comunitária e hospitalar

Escherichia coli (E. coli) constituye una de las principales enterobacterias patógenas causantes de infecciones asociadas a la asistencia en salud, con alto impacto a nivel hospitalario y comunitario por su elevada tasa de morbilidad y mortalidad. El mecanismo de defensa de estas bacterias es mediante la generación de enzimas de resistencia, como la producción de Beta-lactamasas de espectro extendido. OBJETIVO: caracterizar la resistencia a betalactámicos de espectro extendido a partir de su prevalencia en aislados de Escherichia coli de origen comunitario e intrahospitalario de la ciudad de Azogues. MATERIALES Y MÉTODOS: estudio de tipo descriptivo, documental. La población estuvo conformada por 877 registros de la base de datos WHONET de aislados de E. coli procedentes de muestras de origen comunitario e intrahospitalario del laboratorio del Hospital Homero Castanier Crespo. RESULTADOS: el 75,5% de los aislados fueron de mujeres y el 24,5 % de varones. La media de edad fue 43,5 años. La frecuencia de E. coli productora de BLEE fue del 17,7 % con mayor frecuencia en varones (23,7%), en el área de clínica (25,2%), cirugía (16,8%) y en muestras de herida quirúrgica (11,6%). Predominó la resistencia a betalactámicos (84,5%) y cefalosporinas de primera y segunda generación mayor al 48%. Los carbapenemes (imipenen 97,3% y meropenem 96,7%), aminoglucósidos (amikacina 94,9 y gentamicina 80,5), fosfomicina (90,3) y nitrofurantoina (96,7%) mostraron mayor sensibilidad. CONCLUSIONES: la monitorización constante de enzimas BLEE, permite la detección temprana de patrones de sensibilidad y a la vez orienta a un tratamiento terapéutico adecuado, evitando generación de nuevas resistencias y altas tasas de morbilidad y mortalidad.

Escherichia coli (E. coli) is one of the main pathogenic enterobacteria causing infections associated with health care, with high impact at hospital and community level due to its high morbidity and mortality rate. The defense mechanism of these bacteria is through the generation of resistance enzymes, such as the production of extended spectrum beta-lactamases. OBJECTIVE: to characterize extended-spectrum beta-lactam resistance based on its prevalence in Escherichia coli isolates of community and intrahospital origin in the city of Azogues. MATERIALS AND METHODS: descriptive, documentary study. The population consisted of 877 records from the WHONET database of E. coli isolates from community and in-hospital samples from the Homero Castanier Crespo Hospital laboratory. RESULTS: 75.5% of the isolates were from females and 24.5% from males. The mean age was 43.5 years. The frequency of BLEE-producing E. coli was 17.7%, with a higher frequency in males (23.7%), in the clinical area (25.2%), surgery (16.8%) and in surgical wound samples (11.6%). Resistance to beta-lactams predominated (84.5%) and first and second generation cephalosporins was greater than 48%. Carbapenems (imipenem 97.3% and meropenem 96.7%), aminoglycosides (amikacin 94.9 and gentamicin 80.5), fosfomycin (90.3) and nitrofurantoin (96.7%) showed higher sensitivity. CONCLUSIONS: the constant monitoring of BLEE enzymes allows the early detection of sensitivity patterns and at the same time guides to an adequate therapeutic treatment, avoiding the generation of new resistances and high morbidity and mortality rates.

A Escherichia coli (E.coli) é uma das principais enterobactérias patogênicas que causam infecções associadas aos cuidados de saúde, com alto impacto em nível hospitalar e comunitário devido a sua alta taxa de morbidade e mortalidade. O mecanismo de defesa dessas bactérias é através da geração de enzimas de resistência, tais como a produção de beta-lactamases de espectro estendido. OBJETIVO: caracterizar a resistência beta-lactam de amplo espectro com base em sua prevalência em Escherichia coli isolados de origem comunitária e intra-hospitalar na cidade de Azogues. MATERIAIS E MÉTODOS: estudo descritivo, documental. A população consistia de 877 registros do banco de dados WHONET de E. coli isolados de amostras comunitárias e hospitalares do laboratório do Hospital Homero Castanier Crespo. RESULTADOS: 75,5% dos isolados eram de fêmeas e 24,5% de machos. A idade média era de 43,5 anos. A freqüência da E. coli produtora de BLEE foi de 17,7%, com maior freqüência em homens (23,7%), na área clínica (25,2%), na cirurgia (16,8%) e em amostras de feridas cirúrgicas (11,6%). A resistência aos beta-lactâmicos (84,5%) e às cefalosporinas de primeira e segunda geração predominou (mais de 48%). Carbapenems (imipenem 97,3% e meropenem 96,7%), aminoglicosídeos (amikacin 94,9 e gentamicina 80,5), fosfomicina (90,3) e nitrofurantoína (96,7%) mostraram maior sensibilidade. CONCLUSÕES: o monitoramento constante das enzimas BLEE permite a detecção precoce de padrões de sensibilidade e, ao mesmo tempo, orienta para um tratamento terapêutico adequado, evitando a geração de novas resistências e altas taxas de morbidade e mortalidade.

Activity of Ethanolic and Supercritical Propolis Extracts in Corynebacterium pseudotuberculosis and Its Associated Biofilm.

Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis in small ruminants, a chronic disease characterized by the development of granulomas in superficial and visceral lymph nodes as well as in several organs. An important characteristic of the infection with this bacterium is the formation of a biofilm and the absence of effective antibiotic therapy against the disease. From this scenario, the objective of this study was to evaluate the susceptibility of C. pseudotuberculosis to conventional antibiotics and to red, green, and brown propolis extracts obtained by the supercritical and ethanolic extraction methods as well as its activity in the bacterial biofilm. The results of the sensitivity test using antibiotics indicated a sensitivity of C. pseudotuberculosis strains to the antimicrobial agents. The ethanolic extract of green propolis and the supercritical red propolis extract showed the best antibacterial activities against planktonic C. pseudotuberculosis. A lower antimicrobial activity of the brown propolis extract was identified. Propolis extracts were effective in interfering with the formation of the C. pseudotuberculosis biofilm but had little activity on the consolidated biofilm. In conclusion, propolis extracts are more effective against C. pseudotuberculosis in the planktonic stage, being able to interfere with the formation of bacterial biofilm. However, the action of propolis extracts in a sessile and structured microbial biofilm is reduced.

Removal of Mixed-Species Biofilms Developed on Food Contact Surfaces with a Mixture of Enzymes and Chemical Agents.

Sanicip Bio Control (SBC) is a novel product developed in Mexico for biofilms' removal. The aims of this study were to evaluate (i) the removal of mixed-species biofilms by enzymatic (protease and α-amylase, 180 MWU/g) and chemical treatments (30 mL/L SBC, and 200 mg/L peracetic acid, PAA) and (ii) their effectiveness against planktonic cells. Mixed-species biofilms were developed on stainless steel (SS) and polypropylene B (PP) in whole milk (WM), tryptic soy broth (TSB) with meat extract (TSB+ME), and TSB with chicken egg yolk (TSB+EY) to simulate the food processing environment. On SS, all biofilms were removed after treatments, except the enzymatic treatment that only reduced 1-2 log10 CFU/cm2, whereas on PP, the reductions ranged between 0.59 and 5.21 log10 CFU/cm2, being the biofilms developed in TSB+EY being resistant to the cleaning and disinfecting process. Higher reductions in microbial load on PP were reached using enzymes, SBC, and PAA. The employed planktonic cells were markedly more sensitive to PAA and SBC than were the sessile cells. In conclusion, biofilm removal from SS can be achieved with SBC, enzymes, or PAA. It is important to note that the biofilm removal was strongly affected by the food contact surfaces (FCSs) and surrounding media.

Silver nanoparticles obtained from Brazilian pepper extracts with synergistic anti-microbial effect: production, characterization, hydrogel formulation, cell viability, and in vitro efficacy.

The synthesis of silver nanoparticles using plant extracts is known as a green approach, as it does not require the use of high pressure, energy, high temperature, or toxic chemicals. The approach makes use of plant extracts in a process called bioreduction, which is mediated by enzymes, proteins, amino acids, and metabolites found in bark, seed, and leaf extracts, transforming silver ions into metallic silver. This work aimed at developing silver nanoparticles (AgNPs) from Brazilian pepper, applying this green methodology. Hydroalcoholic extract of leaves of Schinus terebinthifolius Raddi was prepared and its concentration of polyphenols, tannins, and saponins quantified. The produced nanoparticles were characterized by UV-Vis spectroscopy, thermogravimetric analysis (TG), dynamic light scattering (DLS), and zeta potential (ZP). AgNPs were formulated in sodium alginate hydrogels to obtain a nano-based semi-solid formulation for skin application. The obtained silver nanoparticles of mean size between 350 and 450 nm showed no cytotoxicity against L929 mouse fibroblasts within the concentration range of 0.025 mg/mL and 10 mg/mL. Schinus terebinthifolius Raddi was found to enhance microbial inhibition against the tested strains, especially against gram-negative bacteria. Its potential use as an alternative to overcome bacterial resistance can be expected.

Antimicrobial resistance of Escherichia coli isolates from spray‑chilled sheep carcasses during cooling / Resistencia antimicrobiana en aislados de Escherichia coli de canales de ovejas asperjadas durante la refrigeración / Resistência antimicrobiana de isolados de Escherichia coli de carcaças ovinas aspergidas durante o resfriamento

Abstract Background: Multidrug-resistant bacteria present in food of animal origin raise human and animal health concerns. Objective: To assess antimicrobial resistance of Escherichia coli isolates from sheep carcasses subjected to spray-chilling with water (4 and 10 hours) during cooling. Methods: Thirty surface swabs were collected from carcasses before and after the last water spray in two slaughter periods. In a first assessment (1st sampling), three spray-chilled carcasses (4 hours), three non-sprayed and one control carcass were sampled. In a second assessment (2nd sampling), the same number of carcasses and treatments were maintained, but spray-chilling was extended to 10 hours. All samples collected were isolated and submitted to susceptibility test using 16 (1st sampling) and 17 (2nd sampling) antimicrobials, respectively. Results: Overall, E. coli isolates were resistant most antimicrobials. Spray-chilled and control carcasses (10 hours) showed resistance to meropenem. Conclusion: E. coli isolates from carcasses subjected to spray-chilling with water for 10 hours had higher antimicrobial resistance to one, two, and four antimicrobial classes, characterizing a multidrug resistance profile. These results highlight the need to monitor health status throughout the meat production processes.

Resumen Antecedentes: las bacterias multirresistentes presentes en alimentos de origen animal son motivo de alerta para la salud humana y animal. Objetivo: verificar la resistencia a antimicrobianos de aislados de Escherichia coli en canales ovinas sometidas a aspersión (4 y 10 h) durante la refrigeración. Métodos: Luego de dos faenas de sacrificio, treinta hisopos fueron colectados en la superficie de las canales antes y después de la última aspersión. En un primer sacrificio (1era colecta) se recolectaron muestras de tres canales sometidas a aspersión (4 horas), tres sin aspersión y una canal como control. En un segundo sacrificio (2da colecta), el mismo número de canales y tratamientos se mantuvo, y el período de aspersión se extendió a 10 horas. Las muestras recogidas fueron aisladas y sometidas a la prueba de susceptibilidad utilizándo 16 (1.ª colecta) y 17 (2.ª colecta) antimicrobianos, respectivamente. Resultados: los aislamientos de E. coli fueron, en general, resistentes a las principales clases de antimicrobianos. Las canales con aspersión y el control (10 h) presentaron resistencia al meropenem. Conclusión: cuando la asperción duró 10 h, los aislados de E. coli presentaron mayor resistencia para una, dos y cuatro clases de antimicrobianos, es decir, fueron multirresistentes a los fármacos utilizados. Esto resalta la necesidad de monitorear el estado de salud durante todos los procesos de producción de carne.

Resumo Antecedentes: bactérias multirresistentes presentes em alimentos de origem animal são motivo de preocupação e alerta na saúde humana e animal. Objetivo: verificar a resistência antimicrobiana em isolados de Escherichia coli de carcaças de ovinos pulverizadas ou não (4 e 10 horas) durante a refrigeração. Métodos: foram coletados trinta swabs de superfície em carcaças antes e após a última aspersão em dois abates. Em outubro do 2015, três carcaças aspergidas foram amostradas, três sem aspersão e uma carcaça para controle, por um período de 4 horas. Em julho de 2016 (2ª coleta), o mesmo número de carcaças e tratamentos foram mantidos e o período de aspersão foi prolongado em 10 horas. As amostras coletadas foram isoladas e submetidas ao teste de susceptibilidade em 16 (1ª coleta) e 17 (2ª coleta) antimicrobianos, respectivamente. Resultados: isolados de E. coli foram, em geral, resistentes às principais classes de antimicrobianos. As carcaças e o controle aspergidos (10 h) apresentaram resistência ao meropenem. Conclusão: quando a aspersão de água durou 10 horas, os isolados de E. coli apresentaram maior resistência antimicrobiana a uma, duas e quatro classes de antimicrobianos, o que é uma multirresistência aos fármacos testados. Isso alerta para a necessidade de monitorar os aspectos de saúde durante todos os processos de produção de carne.

Melanin pigmented gingival tissue impairs red-light lateral scattering for antimicrobial photodynamic therapy.

Gingival melanin pigmentation is present in many African and Oriental descendant people and its occurrence in patients may interfere with the absorption and scattering of therapeutic doses of light. Antimicrobial photodynamic therapy (aPDT) is used as an adjunctive treatment for periodontitis and light irradiation may be impaired by tissue size and its melanin content. The aim of this clinical study was to measure the red-light attenuation in gingival tissue naturally pigmented with melanin. Ten patients with melanized gingival tissue were selected and irradiated by 100 mW red laser. The patients were photographed in frontal and incisal regions with a T2i camera (Canon, Japan) with 100 mm macro lens, 35 mm focal length, aperture f22, 1/100 shutter speed and ISO 200. Three randomly selected sites of each patient were used for evaluations and the irradiation values were assessed in the IMAGEJ software (NIH, Wayne Rasband, USA). Intensity in pixels was quantified in relation to the distance from the light incident point. Data were normalized and the results were presented as relative light intensity as a function of distance. The results demonstrated that red laser light is exponentially attenuated as a function of lateral distance and loses approximately 50 % of its intensity by 2.23 mm. On the other hand, the light travels 3 mm in depth to decay 50 %. In conclusion, our data suggest that melanin presence decreases optical pathway and irradiation protocols for antimicrobial photodynamic therapy in gingival tissue should consider light attenuation and depth of periodontal pockets so that efficient illumination of the target tissue occurs. Periodontal pockets bigger than 6 mm should be irradiated with more than one point.

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.

Subinhibitory concentrations of silver nanoparticles and silver nitrate on the adaptative and cross-resistance to antibiotics on bovine mastitis pathogens / Concentrações subinibitórias de nanopartículas de prata e nitrato de prata na resistência adaptativa e cruzada com antibióticos em patógenos de mastite bovina

Biocides and/or antibiotics used in subinhibitory concentrations can promote the development of adaptive resistance or even cross-resistance in microorganisms. However, studies on these responses following silver treatments are scarce in the literature. Silver-based compounds, including silver nanoparticles (Ag-NPs), can be an alternative in the prevention and treatment of bovine mastitis. Thus, this research evaluated the effect of subinhibitory dosages of Ag-NPs and Ag+ ions from silver nitrate (AgNO3) on Staphylococcus aureus and Escherichia coli isolated from milk of cows with mastitis. Ag-NPs were synthesized by chemical reduction using AgNO3 and sodium citrate and the minimum inhibitory concentration (MIC) of Ag-NPs and Ag+ ions on the mastitis pathogens were determined. Isolates were exposed to subinhibitory concentrations of Ag-NPs or AgNO3 for 10 consecutive days to verify the development of adaptive resistance evaluated by changes in the MIC values. The development of cross-resistance with antibiotics was also studied, being verified by comparing the sensitivity profile of treated cells with non-treated cells. AgNO3 was more effective against all isolates. There was no change in the MIC values or in the antibiotic sensitivity profile for both bacteria following consecutive exposure to subinhibitory dosages of Ag-NPs or AgNO3, indicating that silver was not able to select adaptive resistance or cross resistance to the tested antibiotics. The potential of silver presented by these results is favorable to the continuity of studies aiming to elaborate silver-based therapies for the treatment of bovine mastitis.(AU)

Biocidas e/ou antibióticos em concentrações sub-inibitórias podem promover o desenvolvimento de resistência adaptativa ou mesmo resistência cruzada nos micro-organismos. Entretanto, estudos destas respostas após o tratamento com a prata são escassos na literatura. Compostos a base de prata, incluindo as nanopartículas de prata (Ag-NPs), podem ser uma alternativa na prevenção e/ou tratamento de mastite bovina. Assim, este trabalho objetivou determinar o efeito de doses sub-inibitórias de Ag-NPs e dos íons Ag+ provenientes do nitrato de prata (AgNO3) sobre isolados de Staphylococcus aureus e de Escherichia coli, provenientes de leite de vacas com mastite. As Ag-NPs foram sintetizadas por redução química utilizando AgNO3 e citrato de sódio e a Concentração Mínima Inibitória (CMI) das Ag-NPs e íons Ag+ nos patógenos da mastite foi determinada. Os isolados foram expostos a concentrações sub-inibitórias de Ag-NPs ou de AgNO3 por 10 dias consecutivos para verificar o desenvolvimento de resistência adaptativa à prata pela mudança no valor da CMI, e de resistência cruzada com antibióticos pela mudança no perfil de sensibilidade em relação ao controle. AgNO3 apresentou-se mais efetivo contra todos os isolados. Não foi verificada alteração no valor da CMI nem do perfil de sensibilidade aos antibióticos, indicando que não houve seleção de resistência adaptativa à prata e de resistência cruzada aos antibióticos pelos micro-organismos estudados. O uso potencial da prata apresentado nos resultados é favorável à continuidade dos estudos objetivando a elaboração de terapias à base de prata para o tratamento da mastite bovina.(AU)

Bactérias resistentes isoladas de superfícies inanimadas em um hospital público / Bacterias resistentes aisladas de superficies inanimadas en un hospital público / Isolated resistant bacteria from inanimate surfaces in a public hospital

RESUMO Objetivo: descrever o perfil de resistência a antimicrobianos de microrganismos presentes em superfícies inanimadas. Método: estudo descritivo, realizado entre fevereiro e junho de 2018. Coletou-se 40 amostras microbiológicas de superfícies da Clínica Médica e Unidade de Terapia Intensiva Adulto em um hospital de Mato Grosso, Brasil. A identificação e sensibilidade microbiana foram realizadas através do VITEK-2. A análise dos resultados de resistência foi avaliada conforme as diretrizes do Clinical Laboratory Standards Institute. Resultados: foram isolados 32 microrganismos das 22 amostras contaminadas, dentre eles 14 (43,8%) Staphylococcus coagulase negativa, sete (21,9%) Acinetobacter baumanni complex, três (9,4%) Enterobacter aerogenes. Dentre os Staphylococcus coagulase negativa, 11 (78,6%) apresentaram multirresistência a antimicrobianos e três (42,9%) dos isolados de Acinetobacter baumanni complex foram extremamente resistentes. Conclusão: evidenciou-se a necessidade de educação com ênfase na desinfecção correta e frequente de superfícies e na higienização das mãos após tocar o paciente e as superfícies próximas a ele.

RESUMEN Objetivo: describir el perfil de resistencia de microrganismos presentes en superficies inanimadas a agentes antimicrobianos. Método: estudio descriptivo realizado entre febrero y junio de 2018. Se recolectaron 40 muestras microbiológicas de superficies de la Clínica Médica y de la Unidad de Cuidados Intensivos para Adultos de un hospital de Mato Grosso, Brasil. Los procesos de identificación y sensibilidad microbiana se realizaron a través del dispositivo VITEK 2. El análisis de los resultados de resistencia se evaluó conforme a las directrices del Clinical Laboratory Standards Institute. Resultados: se aislaron 32 microrganismos de las 22 muestras contaminadas; entre ellos, hubo 14 (43,8%) Staphylococcus coagulasa negativa, siete (21,9%) Acinetobacter baumanni complex y tres (9,4%) Enterobacter aerogenes. Entre los Staphylococcus coagulasa negativa, 11 (78,6%) presentaron multi-resistencia a agentes antimicrobianos y tres (42,9%) de los aislados bacterianos de Acinetobacter baumanni complex fueron extremamente resistentes. Conclusión: se hizo evidente la necesidad de instrucción con énfasis en la correcta y frecuente desinfección de superficies y en el lavado de manos después de entrar en contacto con el paciente y con las superficies próximas al paciente.

ABSTRACT Objective: to describe the antimicrobial resistance profile of microorganisms present on inanimate surfaces. Method: a descriptive study, conducted between February and June 2018. Forty microbiological samples were collected from surfaces of the Medical Clinic and Intensive Care Unit for Adults in a hospital located in Mato Grosso, Brazil. Microbial identification and sensitivity were performed by means of VITEK 2. The analysis of the resistance results was assessed according to the Clinical Laboratory Standards Institute guidelines. Results: a total of 32 microorganisms were isolated from the 22 contaminated samples, the following among them: 14 (43.8%) Coagulase-Negative Staphylococcus, seven (21.9%) Acinetobacter baumanni complex, and three (9.4%) Enterobacter aerogenes. Of the Coagulase-Negative Staphylococcus, 11 (78.6%) presented multi-drug resistance to antimicrobial agents, and three (42.9%) of the Acinetobacter baumanni complex isolates were extremely resistant. Conclusion: this study evidenced the need for education with emphasis on proper and frequent disinfection of surfaces and on hand hygiene after touching patients and surfaces close to them.