"Antibacterial effect and possible mechanism of action of 1,3,4-oxadiazole in Staphylococcus aureus".

Staphylococcus aureus is one of the main etiological agents causing foodborne diseases, and the development of new antibacterial agents is urgent. This study evaluated the antibacterial activity and the possible mechanism of action of the 1,3,4-oxadiazole LMM6 against S. aureus. The minimum inhibitory concentration (MIC) of LMM6 ranged from 1.95 to 7.81 µg ml-1. The time-kill assay showed that 48-h treatment at 1× to 8× MIC reduced S. aureus by 4 log colony forming unit (CFU), indicating a bacteriostatic effect. Regarding the possible mechanism of action of LMM6, there was accumulation of reactive oxygen species (ROS) and an increase in the absorption of crystal violet (∼50%) by the cells treated with LMM6 at 1× and 2× MIC for 6-12 h. In addition, there was increased propidium iodide uptake (∼84%) after exposure to LMM6 for 12 h at 2× MIC. After 48 h of treatment, 100% of bacteria had been injured. Scanning electron microscopy observations demonstrated that LMM6-treated cells were smaller compared with the untreated group. LMM6 exhibited bacteriostatic activity and its mechanism of action involves increase of intracellular ROS and disturbance of the cell membrane, which can be considered a key target for controlling the growth of S. aureus.

Comportamento da população em relação aos cuidados com os alimentos durante a pandemia da COVID-19 / Population behavior in relation to food care during the COVID-19 pandemic

Este estudo analisou o conhecimento e comportamento em relação aos cuidados e higiene alimentar antes e durante a pandemia da COVID-19 no Brasil. Os participantes foram recrutados pelas redes sociais para responder um questionário sobre aspectos sociodemográficos, conhecimento sobre coronavírus, isolamento social e recebimento de informações sobre higienização de alimentos e suas embalagens. Participaram da pesquisa 1.061 indivíduos, sendo a maioria do sexo feminino (87%), com até 35 anos (69,9%); 82,8% tinham ou estavam a concluir o ensino superior; e a renda mensal de 63,2% era de até 6 salários mínimos. Sobre a higiene de frutas e hortaliças, 56,59% dos participantes passaram a usar água e sabão durante a pandemia. Quanto a limpeza das embalagens dos alimentos recebidos por delivery, 71,85% dos participantes passaram a limpar as embalagens durante a pandemia. De forma geral, pode-se observar modificações significativas nos cuidados com os alimentos durante a pandemia da COVID-19.

This study during knowledge and behavior in food care and hygiene before and that of COVID-19 in Brazil. Participants were recruited through social networks for respondents on the sociodemographic aspects of the population's knowledge about coronavirus, social isolation and receiving information on hygiene of food and its packaging. A total of 1,061 participated in the survey, the majority being female (87%), aged up to 35 years (69.9%); 8.8% had or 2.8% had higher education; and 63.6% monthly income was up to 6% monthly. Regarding the hygiene of fruits and vegetables, 56.59% of the participants chose soap and water during the. As for cleaning packages received by delivery, 71% of patients choose to clean packages during the pandemic. In general, the pandemic can be considered in the care with food of COVID-19.

Synergistic activity of Stryphnodendron adstringens and potassium sorbate against foodborne bacteria.

Stryphnodendron adstringens is a medicinal plant that has a broad spectrum of action, including antibacterial activity. The aim of the present study was to evaluate the effect of S. adstringens alone and in combination with potassium sorbate (PS) against foodborne bacteria. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined and, for most of the bacteria tested, the crude extract (CE), aqueous fraction (AQF), and ethyl-acetate fraction (EAF) of S. adstringens had a MIC and MBC ranging from 500 to ≥ 1000 µg/mL. The AQF and EAF showed greater activity against S. aureus strains (MIC = 125 to 250 µg/mL; MBC = 500 to 1000 µg/m). Quantitative cell viability was determined and was observed reductions ranging from 3.0 to 5.8 log10 CFU/ml.The combination of S. adstringens and PS against seven S. aureus isolates was determined by the checkerboard method at neutral and acid pH. In a neutral medium, the AQF + PS combination presented synergistic or additive interactions against six S. aureus strains. The combination of EAF + PS resulted in additive interactions against four bacterial isolates. In an acidic medium, the AQF + PS combination was synergistic or additive against all S. aureus, while EAF + PS presented the same effect against six S. aureus strains S. adstringens showed important antibacterial effects against foodborne S. aureus strains. Moreover, the combination of S. adstringens fractions and PS improved the antibacterial activity compared to the compounds utilized individually. The combined use of these compounds may be an alternative to reduce bacterial food contamination and improve food safety.

Systematic review of disulfiram as an antibacterial agent: what is the evidence?

The objective of this systematic review was to retrieve and examine published studies related to in vitro and in vivo evaluation of disulfiram for the treatment of bacterial infections. Five scientific databases (PubMed, Embase, Scopus, Web of Science, and Latin American and Caribbean Health Sciences Literature) were searched to retrieve the maximum literature regarding the study's aim. The search strategy retrieved a total of 870 studies, of which 31 were included and 19 approached disulfiram as the primary aim and 12 included it as a secondary finding from other investigational objectives. The evidence pointed out five main aspects of pre-clinical testing regarding disulfiram antibacterial activity, namely spectrum of antimicrobial action, drug combinations, intracellular studies, animal studies and bacterial targets. Findings to emerge from this study are the observed potential of disulfiram as a non-antibiotic drug being proposed as a potential drug to contribute to the treatment of bacterial diseases usually with few treatment alternatives in the context of drug resistance. We evaluated the potency and selectivity of disulfiram, which indeed until now shows potential to be explored for use as an adjunctive chemical to antimicrobial ones. Even with the level of evidence being reserved, the potential of combining disulfiram with other drugs, already used or new to be used for the treatment of mycobacterial diseases, as well as its likely immunomodulatory effect, deserve to be further investigated. Furthermore, the copper-dependent mode of action in Gram-positive bacteria is an alternative to be explored in drug design or repurposing of chemicals.

The ancient Tuberculosis in the novel COVID-19 scenario

Abstract Recently, the world has coped with the challenge of the novel SARS-CoV-2 rapid spreading, causing COVID-19. This scenario has overburdened health systems, forced social isolation, and interrupted some services, changing the way how health assistance is provided. The management of chronic infectious diseases such as tuberculosis is a sensitive matter in times when the control strategies are at risk. In this sense, how could a high burden disease such as tuberculosis affect or be affected when combined with the COVID-19 pandemic? Patients with tuberculosis have a social background and lung impairment that represent risks in the pandemic scenario of another widely transmitted respiratory disease. Thus, even with several questions remaining unanswered, research and public policies should be addressed to control the effects of the current highly contagious COVID-19 without forgetting how it will affect the natural progression of patients suffering from tuberculosis.

PanB over-representation as part of pyrazinamide action: a proteomic insight.

Background: Pyrazinamide (PZA) represents a milestone as a first-line antituberculosis drug due to its sterilizing activity against Mycobacterium tuberculosis. Materials & Methods: The protein changes induced by subinhibitory PZA exposure of M. tuberculosis in acidic pH were evaluated by a proteomic approach. Results: Among the 1059 M. tuberculosis proteins identified, the specific acidification in the culture medium induced the over-representation of MurF (Rv2157c), and its under-representation was induced by 12 h of PZA exposure. PanB (Rv2225) was over-represented at 24 h of PZA exposure. Conclusion: The authors highlight the over-representation of PanB in M. tuberculosis correlates of PZA action in acidic pH, reinforcing the role of the pantothenate pathway as a bacillus drug target to be explored.

Activity of (-)-Camphene Derivatives Against Mycobacterium tuberculosis in Acidic pH.

BACKGROUND: For more than 60 years, the lack of new anti-tuberculosis drugs and the increase of resistant Mycobacterium tuberculosis lineages exhibit a therapeutic challenge, demanding new options for the treatment of resistant tuberculosis. OBJECTIVE: Herein, we determined the (i) activities of (-)-camphene and its derivatives and (ii) combinatory effect with pyrazinamide (PZA) against Mycobacterium tuberculosis in acidic pH and (iii) cytotoxicity on VERO cells. METHODS: The activity of (-)-camphene and its 15 derivatives was determined in M. tuberculosis H37Rv in culture medium at pH 6.0 by Resazurin Microtiter Assay Plate (REMA). The activity and combinatory study of three (-)-camphene derivatives with PZA was carried out on seven multidrugresistant (MDR) clinical isolates by REMA and Checkerboard, respectively. The assay of 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) bromide in VERO cells was used to determine the derivatives' cytotoxicity. RESULTS: Four (-)-camphene derivatives, (4), (5a) (5d) and (5h), showed a reduction in the MIC value at pH 6.0 compared to the MIC detected at pH 6.8 in M. tuberculosis H37Rv and multidrug resistant clinical isolates. Three (-)-camphene derivatives, (4), (5d) and (5h), showed synergistic effect (FICI ≤ 0.5) combined with PZA and were more selective for M. tuberculosis than VERO cell (selective index from 7.7 to 84.2). CONCLUSION: Three (-)-camphene derivatives have shown to be promising anti-TB molecule scaffolds due to their low MIC values in acidic pH against MDR M. tuberculosis clinical isolates, synergism with PZA and low cytotoxicity.

pH interferes in photoinhibitory activity of curcumin nanoencapsulated with pluronic® P123 against Staphylococcus aureus.

Microbial contamination control is a public health concern and challenge for the food industry. Antimicrobial technologies employing natural agents may be useful in the food industry for these purposes. This work aimed to investigate the effect of photodynamic inactivation using curcumin in Pluronic® P123 nanoparticles (Cur/P123) at different pH and blue LED light against Staphylococcus aureus. Bacterial photoinactivation was conducted using different photosensitizer concentrations and exposure times at pH 5.0, 7.2 and 9.0. A mixture design was applied to evaluate the effects of exposure time (dark and light incubation) on the photoinhibitory effect. S. aureus was completely inactivated at pH 5.0 by combining low concentrations of Cur/P123 (7.80-30.25 µmol/L) and light doses (6.50-37.74 J/cm2). According to the mathematical model, dark incubation had low significance in bacterial inactivation at pH 5.0 and 9.0. No effect in bacterial inactivation was observed at pH 7.2. Cur/P123 with blue LED was effective in inactivating S. aureus. The antimicrobial effect of photodynamic inactivation was also pH-dependent.

Hypericin-mediated photoinactivation of polymeric nanoparticles against Staphylococcus aureus.

Photoinactivation is a promising technique for Staphylococcus aureus control. This microorganism causes foodborne diseases (DTAs) and forms biofilms that are highly resistant and difficult to eradicate. Thus, the aim of this study was to evaluate the photodynamic activity of hypericin (HYP) in polymeric nanoparticles (Pluronic® P123) against S. aureus planktonic and biofilm cells. Planktonic cells and biofilms of S. aureus (ATCC 25923) were subjected to photoinactivation using low-power orange LED (0.3 mW/cm²) with different HYP formulation concentrations in Pluronic® P123. The P123 molar ratios were 2.5 (HYP/P123-2.5) and 10 (HYP/P123-10), respectively. The treatment times for planktonic cells were proposed by a mixture design, and bacterial photoinactivation was observed in concentrations of 12.5 to 3.12 µmol/L for HYP/P123-2.5 and reductions of ∼ 4.0 log CFU/mL in 12.5 to 0.78 µmol/L for HYP/P123-10. For biofilms, 30 min of darkness and 30 min of illumination were used. Maximum reductions were similar for both formulations and corresponded to approximately 0.9 log CFU/cm². It was concluded that photoinactivation with longer lighting times was effective against planktonic cells and could be potentially applied to control S. aureus.

Minimum Bactericidal Concentration Techniques in Mycobacterium tuberculosis: A Systematic Review.

Minimum bactericidal concentration (MBC) assay is an accepted parameter for evaluating new antimicrobial agents, and it is frequently used as a research tool to provide a prediction of bacterial eradication. To the best of our knowledge, there is no standardization among researchers regarding the technique used to detect a drug's MBC in Mycobacterium tuberculosis. Thus, the aim of this systematic review is to discuss the available literature in determining a drug's MBC in M. tuberculosis, to find the most commonly used technique and standardize the process. A broad and rigorous literature search of three electronic databases (PubMed, Web of Knowledge, and LILACS) was performed according to the PRISMA statement. We considered studies that were published from January 1, 1990 to February 19, 2019. Google Scholar was also searched to increase the number of publications. We searched for articles using the MeSH terms "microbiological techniques," "Mycobacterium," "antibacterial agents." In addition, free terms were used in the search. The search yielded 6,674 publications. After filter application, 5,348 publications remained. Of these, we evaluated the full text of 187 publications. By applying the inclusion criteria, 69 studies were included in the present systematic review. In the literature analyzed, a great variety in the techniques used to determine a drug's MBC in M. tuberculosis was observed. The most common variability is related to the culture media used, culture incubation time, and the percentage of bacterial death for the drug to be considered as bactericidal. The most commonly used technique for drug's MBC determination was carried out using the drug's minimum inhibitory concentration (MIC) assay. Aliquots from prior MIC values were subcultured in Middlebrook agar and incubated for 4 weeks at 35°C for determining the colony forming unit (CFU) with relevance to detect 99.9% bacilli killed or reduction in 3 log10 viable bacilli.

Serial Systemic Candida albicans Infection Highlighted by Proteomics.

Candida albicans is the major pathogen isolated from nosocomial bloodstream infections, leading to higher mortality rates. Thus, due to its clinical relevance, studies aiming to understand host-pathogen interactions in C. albicans infection are necessary. Therefore, we performed proteomic analysis using a murine model of serial systemic infection by C. albicans to evaluate possible changes in the protein profile of the pathogen over time. Firstly, we observed a reduction in the median survival time of infected animals with increasing passage number, suggesting a higher pathogenicity acquired during repeated infections. By LC-MS/MS, it was possible to obtain protein profiles from the wild-type strain (WT) and compare them to proteins extracted from Candida cells recovered from infected tissues during passages one, three, and four (P1, P3, and P4). We obtained 56, 29, and 97 proteins in P1, P3, P4, respectively, all varying in abundance. Regarding biological processes, the majority of proteins were related to carbohydrate metabolism, stress responses and amino acid metabolism. The proteins were also categorized according to their potential role in virulence traits, such as biofilm production, yeast-to-hyphae transition, phenotypic switching, proteins related to stress responses, and uncharacterized proteins. Therefore, serial infection in combination with proteomic approach enabled us to deepen the existing knowledge about host-pathogen interactions.

Promising Antituberculosis Activity of Piperine Combined with Antimicrobials: A Systematic Review.

Piperine, a bioactive compound from Piper nigrum and Piper longum, has shown promising activity as efflux pump (EP) inhibitor and as adjunct in treatment of tuberculosis (TB). The present systematic review investigated scientific studies of the activity of piperine against mycobacteria, with a focus on its mechanism of action, drug interactions, and antimycobacterial activity. A broad and rigorous literature search of three electronic databases (PubMed, Web of Knowledge, and LILACS) was performed according to the PRISMA statement. We considered studies that were published up to December 1, 2017. Google Scholar was also searched to increase the number of publications. We searched for articles using the search terms "piperine" and "Mycobacterium spp." The search yielded a total of 225 articles. After removing duplicate publications, 208 publications remained. Of these, we evaluated the full text of 13 articles. After applying the inclusion criteria, eight studies were included in the present systematic review. The results of the systematic review showed that piperine has promising anti-TB activity, mainly when combined with antimicrobials, and plays an important role as an EP inhibitor.

Combinatory activity of linezolid and levofloxacin with antituberculosis drugs in Mycobacterium tuberculosis.

SETTING: The increase of multidrug and extensively drug resistant Mycobacterium tuberculosis strains turns the search for new tuberculosis (TB) treatment options of paramount importance. OBJECTIVE: In this sense, the present study evaluates the in vitro activity of isoniazid (INH)/rifampicin (RIF)/levofloxacin (LVX) and INH/RIF/linezolid (LNZ) combinations in resistant M. tuberculosis. DESIGN: The activities of the combinations were evaluated with M. tuberculosis H37Rv, susceptible and 10 resistant clinical isolates by three-dimensional checkerboard. LVX and LNZ were used as the third drug at fixed ½ and » minimum inhibitory concentration (MIC). INH and RIF were tested at concentrations ranging from 0.0009 µg/mL to 50 µg/mL and 0.0009 µg/mL to 800 µg/mL, respectively. The combinatorial effects were determined by the Fractional Inhibitory Concentration Index (FICI). FICI values ≤ 0.75, 0.75-4 and ≥4 were considered as synergism, indifferent and antagonism, respectively. RESULTS: MIC ranged from 0.03 - 6.25 µg/mL for INH, 0.008-100 µg/mL for RIF, 0.12-0.25 µg/mL for LVX and 0.25-0.5 µg/mL for LNZ in the H37Rv and all clinical isolates. INH/RIF/LVX and INH/RIF/LNZ synergisms were observed in 40 and 50% of the resistant M. tuberculosis clinical isolates and better observed for INH and RIF combined to LVX or LNZ at » MIC. CONCLUSION: The present study calls attention for the potential use of INH/RIF/LVX and INH/RIF/LNZ combinations in the treatment of resistant TB.

RDRioMycobacterium tuberculosis lineage in the Brazil/Paraguay/Argentina triple border.

The high tuberculosis (TB) incidence rates, the closeness of the cities and the high migration flux on the Brazil/Paraguay/Argentina border deserves an in-depth study, using Mycobacterial Interspersed Repetitive Unit (MIRU) and Spoligotyping genetic markers to explore the impact of the Mycobacterium tuberculosis RDRio lineage on disease transmission and resistance to anti-TB drugs in this setting. Although without the totality of M. tuberculosis isolates causing TB in this studied setting, a number of 97 isolates obtained from sputa samples culture of patients with confirmed TB, from 2013 to 2015, were submitted to 24 loci MIRU, Spoligotyping, detection of RDRio lineage and detection of mutation related to isoniazid and rifampicin resistance by MTBDRplus/DNA STRIP. In this sample, it was observed high clonal variability of circulating M. tuberculosis isolates causing TB in Brazilian cities bordering Paraguay and Argentina. The percentage of RDRio lineage causing TB in this setting was 15.46%, and lower than the detected in different areas of Brazil. According to 24 loci MIRU, the major MIRU International Type (MIT) related with RDRio lineage were MIT 26, MIT 738, MIT 601 with four, two and one isolates, respectively. Eight isolates with RDRio marker were classified as orphans. The mainly Spoligofamily related with RDRio lineage was LAM1 and LAM9 and no relationship between RDRio lineage and resistance in M. tuberculosis isolates circulating in this setting could be established. This work is pioneer in studying the dynamics of RDRio lineage transmission on the Brazil/Paraguay/Argentina border and deserves further studies to analyze the real contribution of the RDRio lineage in outbreaks and the risk of significant development of MDR-TB in the setting studied.

Antibacterial and antibiofilm activity of carvacrol against Salmonella enterica serotype Typhimurium

ABSTRACT The present study evaluated the antibacterial and antibiofilm activity of carvacrol against Salmonella Typhimurium. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined and the time-kill curve and scanning electron microscopy (SEM) were performed to evaluate antibacterial activity. Antibiofilm activity was evaluated by quantifying total biomass using crystal violet assay, and metabolic activity was determined using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The action of carvacrol against preformed biofilm on polypropylene and stainless steel was also evaluated by colony counting and SEM. The MIC and MBC was 312 µg mL-1. Carvacrol at MIC and 2 x MIC eliminated cells after 6 and 1 h of treatment, respectively, as exhibited in the time-kill curve. The greatest reduction in biofilm biomass and metabolic activity was 1,719 OD550 and 0,089 OD550 respectively, both at 4 x MIC of carvacrol. In carvacrol treated biofilms of S. Typhimurium on polypropylene, a reduction of 5.12 log was observed with 4 x MIC, while on stainless steel, carvacrol at 4 x MIC reduced bacterial counts by 5 log. The results showed that carvacrol exhibits antibacterial activity and can be used as an alternative for the control of S. Typhimurium biofilms.

Systematic review on the proteomic profile of Mycobacterium tuberculosis exposed to drugs.

The authors present an overview about proteomics studies in Mycobacterium tuberculosis exposed to some anti-tuberculosis drugs and new candidates, using two-dimensional gel electrophoresis and mass spectrometry. To date, that the authors have knowledge, this is the first studies that was performed specifically in M. tuberculosis using systematic review on electronic literature conducted in three databases using the following search terms: tuberculosis OR mycobacterium tuberculosis, proteome OR proteomics, and mass spectrometry electrospray ionization OR matrix-assisted laser desorption ionization OR two-dimensional gel electrophoresis. By electronic search, 622 abstracts of the original articles published from November 2003 to March 2016 were selected. After the selection, four articles fulfill proposed criteria and were included in this study. The studies reported changes in the protein profile of M. tuberculosis after exposure to isoniazid, ethambutol, streptomycin, ofloxacin, moxifloxacin and two new drugs candidates, SQ109 and ATB107. In conclusion, the proteins changes were related to the synthesis of mycolic acids, cellular metabolism pathways, bacterial stress and starvation.

Mutations in catalase-peroxidase KatG from isoniazid resistant Mycobacterium tuberculosis clinical isolates: insights from molecular dynamics simulations.

The current multidrug therapy for tuberculosis (TB) is based on the use of isoniazid (INH) in combination with other antibiotics such as rifampin, ethambutol and pyrazinamide. Literature reports have shown that Mycobacterium tuberculosis, the causative agent of TB, has become resistant to this treatment by means of point mutations in the target enzymes of these drugs, such as catalase-peroxidase (KatG). By means of equilibrium molecular dynamics in the presence of the ligand, this work evaluated ten point mutations described in the enzyme KatG that are related to resistance to INH . The results showed that the resistance mechanism is related to stereochemical modifications at the N-terminal domain of the protein, which restrict INH access to its catalytic site, not involving mechanisms of electrostatic nature. These results show insights that can be useful for the identification of new anti-TB drugs which may be able to circumvent this mechanism of resistance.

Prevalence and predictors associated with intestinal infections by protozoa and helminths in southern Brazil.

Approximately 2 billion people are infected with soil-transmitted helminths worldwide, mainly in tropical and subtropical areas. This research aimed to investigate the prevalence and predictors associated with parasitic infections in primary health care. A cross-sectional study was performed with a large random sample to identify the prevalence and predictors associated with parasitic infections in primary health care in Marialva, southern Brazil, from April 2011 to September 2013. Stool samples from 775 individuals were analyzed for the presence of protozoan cysts, helminth eggs, and larvae. The overall prevalence of intestinal parasites was 13.94 %, and the prevalence of protozoa and helminths was 15.1 and 2.9 %, respectively. The predictor variables that were associated with intestinal parasites were male gender odds ratio (OR) 1.60, 95 % confidence interval (CI 1.10-2.40) and the absence of a kitchen garden (OR 2.28, 95 % CI, 1.08-4.85). Positive associations were found between Giardia duodenalis and individuals aged ≤18 with high risk (OR 19.0, 95 % CI 2.16-167.52), between Endolimax nana and the absence of a kitchen garden (p < 0.01), and between Trichuris trichiura and the presence of a kitchen garden (p = 0.014). Polyparasitism was present in 27.27 % of infected individuals. Our findings confirmed a relatively low prevalence in primary care, compared to international standards, despite the rare publications in the area. As variables, male gender and the absence of a kitchen garden stood out as important predictors. It is highly relevant that the health conditions of the population comply with consistent standards.