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Nowadays whole world blessed with medicinal plants for using as a major bio-source of modern synthetic drugs which are used widely in medicinal field. In this study Qualitative experiments of phytochemical and antibacterial tests were performed of ethanolic extracts of leaf and root parts ofHeliotropium supinum. The Qualitative experiments revealed that the presence phytochemicals such as alkaloids, phenolic compounds, tannins, flavonoids, saponins, terpenoids, steroids, glycosides, and carbohydrates. In this experiment alkaloids, phenolic compounds, tannins, flavonoids, saponins, terpenoids, Carbohydrates and glycosides were observed in ethanolic extract leaves while alkaloids, flavonoids terpenoids were found in roots extract. For being presence of these essential propertiesH. supinumshowed antibacterial activity. Maximum zone of inhibition was recorded in 500 µg concentration of leaves and root extracts where leaves extract showed highest inhibition zone (21 mm) againstS. aureuswhile root extracts showed against E. coli (17 mm). Present study was designed to identify the sources and accurate information regarding the active components of this medicinal plant and to enhance awareness to public and private sector.

The antibiotic norfloxacin was adsorbed on clay and carbon-clay nanocomposites (MD- 210 and MDac3-210-500, synthesized by hydrothermal carbonization of dextrose) but maintained its antimicrobial activity once adsorbed. To degrade it, direct (DP) and indirect (IP, with sulfate radicals) photolysis were performed. Antimicrobial activity tests were performed to evaluate the degradation generated. There were no significant differences between DP and IP for unabsorbed norfloxacin solution. The IP treatment caused a loss of antibiotic activity in all cases. For DP, the carbon samples showed better results than those of clay, evidencing the importance of oxygenated carbonaceous groups in the photolysis of norfloxacin., Centro de Tecnología de Recursos Minerales y Cerámica, Centro de Investigación y Desarrollo en Fermentaciones Industriales, Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas, Centro de Investigaciones Ópticas

Armansyah T, Siregar TN, Suhartono, Sutriana A. 2022. Phytochemicals, characterization and antimicrobial tests of red betel leaves on three solvent fractions as candidates for endometritis phytotherapy in Aceh cattle, Indonesia. Biodiversitas 23: 2111-2117. The use of antibiotics to treat endometritis in cattle carries with it some weaknesses in the form of toxicity risks and unexpected side effects. The use of medicinal plants as antimicrobials and immunomodulators has the potential to be an alternative therapy. Red betel leaves could be considered viable candidates for phytotherapy in cattle with endometritis. This study aimed to determine the differences in biological activity and antibacterial ability of red betel leaf through the use of three different solvents, namely n-hexane, ethyl acetate, and ethanol. The highest yield of betel leaf extract was obtained from the ethyl acetate solvent fraction consisting of flavonoids, terpenoids, steroids, tannins, phenolics, and saponins. The highest biological compounds in red betel leaf identified by GC-MS in n-hexane solvent, ethyl acetate, and ethanol were 5-isobutylidene-N, N-dimethylbarbituric acid, benzenamine, 4,4' - (1,2-ethenediyl) bis-, and 9-octadecadienoic acid, respectively. Among the three solvent fractions of red betel leaf extract, it was observed that ethyl acetate exhibited higher antibacterial activity than hexane and ethanol. Hence, it can be concluded that ethyl acetate extract of red betel leaves solvent has secondary metabolites and better antibacterial potential than ethanol and n-hexane solvents.

The applications of organosulfur compounds, especially thioketones are varied which concerns both synthetic and biological chemistry. In this context, from the essential oil of Artemisia judaica L., extracted by hydrodistillation, and consisting mainly of piperitone (91.77 %), thione, and thiol derivatives were hemi-synthesized via a thionylation reaction byphosphorus pentasulphide (P2S5). The possible thione-thiol tautomerism in solution followed spectrophotometrically revealed the predominance of thiol form in solution. The antimicrobial activity of essential oils showed that the thionylation of essential oil caused a considerable increase in antimicrobial activity in particular against Escherichia coli. This work is licensed under a Creative Commons Attribution 4.0 International License., Iz eteričnog ulja biljke Artemisia judaica L., koje se uglavnom sastoji od piperitona (91,77 %), a dobiveno je hidrodestilacijom, reakcijom tionilacije pomoću fosforova pentasulfida (P2S5) hemisintetizirani su tion i tiol derivati. Moguća tion-tiolna tautomerija praćena je spektrofotometrijski. Analiza je pokazala da u otopini dominira tiolni oblika. Također, pokazalo se da tionilacija eteričnog ulja uzrokuje znatno povećanje antimikrobne aktivnosti, osobito kad je u pitanju bakterija Escherichia coli. Ovo djelo je dano na korištenje pod licencom Creative Commons Imenovanje 4.0 međunarodna.

Benzothiazolyl hydrazones are synthesized and tested as hypoglycemic and antimicrobial agents. Condensation of 2-acyl-(1H)-indene-1,3(2H)-diones with 2-hydrazinylbenzo[d]thiazole/2-hydrazinyl-6-substitutedbenzo[d]thiazoles gives the corresponding hydrazones in high yields. Preliminary biological assay of the products reveals significant antidiabetic and antimicrobial activities. The α-amylase and α-glucosidase inhibition assay is used for determination of Type II diabetes inhibitory activity.

Due to the globally increasing need for utilizing antimicrobial materials, it is necessary to improve and develop newer methods and techniques for determining the antimicrobial properties of these materials qualitatively and quantitatively, especially in the medical field. Recently there has been a debate among scientists between the difference between antimicrobial test and sterility test. For antimicrobial and microbial resistance, the test is limited to the standard methods to determine the effect of the sample as an antimicrobial without paying attention to the fact that the sample is sterile or not, unless the sample was taken into sterile atmosphere and isolated with a protective suitable sterile package cover after treatment with the antimicrobial agent because the external atmosphere contains a lot of scattered types of bacterial and fungal microorganisms. In case of sterility test, the sample to be examined should be prepared, isolated and coated from the outside atmosphere, where the presence of microbes on the surface of the sample by standard methods is detected and in some sterilization tests it is prohibited to include an antimicrobial substance to the sample to be tested to avoid the interference with the test. In both cases the tests should be implemented in a sterilized room and conditions according to the recognized scientific principles. In conclusions, the antimicrobial test is used to make sure that the specimen is attained antimicrobial properties or not and the sterilization test is done by ensuring that the sample is free of contaminated microorganisms. This review poses on some factor and conditions affecting antimicrobial action and some standard test methods for determination of antimicrobial and sterility potential of materials.

The application of titanium oxide nanotubes for the removal of contaminants from freshwater is a rapidly growing scientific interest, especially when it comes to water conservation strategies. In this study we employed four different titanium oxide nanotube surfaces, prepared by a two-electrode anodic oxidation. Two of the surfaces were synthesised in aqueous media, while the other two surfaces were synthesised in ethylene glycol. One of the arrays synthesised in the organic medium was impregnated with silver nanoparticles, while the remaining surfaces were not. The chemical reactivity of the various surfaces was assessed using 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,20 -azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as free electron sensitive probe molecules, in parallel with tannic acid degradation and copper ion reducing capacity. The potential antimicrobial activity of the surfaces was assessed against a panel of microorganisms composed of yeast, fungi, Gram-positive and Gram-negative bacteria. Field emission scanning electron microscopy revealed that surfaces produced in the aqueous medium had a smaller tube length and a smaller tube diameter. It was noted that one of the materials using sodium sulfate as the supporting electrolyte had the most irregular nanostructure morphology with tubes growing to the side rather than vertically. The structural variation of the surfaces directly reflected both the chemical and biological activity, with the nanotubes formed in ethylene glycol showing the fastest rates in the stabilization of DPPH and ABTS radicals, the fastest tannic acid decomposition under various pH conditions and the fastest metal reducing activity. Furthermore, the surface containing silver and its bare counterpart showed the most effective, peer-reviewed

Objectives To evaluate the performance of two rapid antimicrobial susceptibility testing (RAST) methods to determine ceftazidime/avibactam susceptibility directly from blood cultures (BCs). Methods A total of 246 Escherichia coli or Klebsiella pneumoniae isolates were tested for ceftazidime/avibactam susceptibility directly from BC bottles using EUCAST RAST and Etest® RAST. Results obtained after 4, 6 and 8 h of incubation were compared with those obtained by reference broth microdilution on pure overnight subcultures. Results In total, the proportion of readable zones after 4 h of incubation was 96.7% and reached 100% after 6 and 8 h of incubation. EUCAST RAST yielded >98% of categorical agreement (CA) with all reading times. Major error (ME) and very major error (VME) rates were inferior to 3%, for each of the reading times. The proportion of results in the area of technical uncertainty (ATU) was almost similar (3.8%–4.1%) at the different reading times. DET-RAST yielded 97.5%, 98% and 99.6% of CA with readings at 4, 6 and 8 h, respectively. One (0.6%) ME was observed at each reading time, whereas five (5.9%) and four (4.5%) VMEs were observed analysing readings at 4 and 6 h, respectively. No VME was observed with readings at 8 h. Conclusions EUCAST RAST was accurate to determine ceftazidime/avibactam susceptibility of carbapenemase-producing K. pneumoniae and E. coli directly from BC bottles. DET-RAST has the advantage of determining MIC values and avoiding ATU results but showed to be an accurate method only with reading at 8 h.

Most bacteria in nature exist in multispecies communities known as biofilms. In the natural habitat where resources (nutrient, space, etc.) are usually limited, individual species must compete or collaborate with other neighboring species in order to perpetuate in the multispecies community. The human oral cavity is colonized by700 microbial species known as the indigenous microbiota. This indigenous flora normally maintains an ecological balance through antagonistic as well as mutualistic interspecies interactions. However, environmental perturbation may disrupt this balance, leading to overgrowth of pathogenic species which could in turn initiate diseases such as dental caries (tooth decay) and periodontitis (gum disease). Understanding the mechanisms of diversity maintenance may help developing novel approaches to manage these "polymicrobial diseases". In this chapter, we will focus on a well-characterized form of biochemical warfare: bacteriocins produced by Streptococcus mutans, a primary dental caries pathogen, and hydrogen peroxide (H

Globally, piperacillin-tazobactam resistance among Escherichia coli and Klebsiella pneumoniae is driven by OXA-1 beta-lactamases. Expression of bla(OXA-1) yields piperacillin-tazobactam MICs of 8 to 16 μg/mL, which straddle the susceptible/susceptible-dose dependent breakpoint set by the Clinical and Laboratory Standards Institute in 2022. Variability of the reference broth microdilution method (BMD) was evaluated by manufacturing BMD panels using 2 brands of piperacillin, 2 brands of tazobactam and 2 brands of cation-adjusted Mueller-Hinton broth. In addition, ETEST, which harbors an intermediate dilution of 12 μg/mL was evaluated for the ability to differentiate isolates with and without bla(OXA-1). A collection of 200 E. coli and K. pneumoniae, of which 82 harbored a bla(OXA-1) gene, were tested. BMD variability was on average 1.3-fold, within the accepted 2-fold variability of MICs. However, categorical agreement (CA) between BMD reads was 74.0% for all isolates and 63.4% for those with a bla(OXA-1) gene and 81.3% for those without bla(OXA-1) detected (P = 0.004, Pearson’s Chi Square). ETEST overall CA with the BMD mode was 68.0% and essential agreement (EA) was 80.5%. For isolates with bla(OXA-1), CA was 50.0% and EA was 69.5%, versus 80.5% and 88.1%, respectively, for isolates without bla(OXA-1) (P

Objective Fibrin sealants have been used for hemostasis, sealant for cerebrospinal fluid leakage, and adhesive barrier in neurosurgery. Further, as its clinical use and role of an effective drug delivery vehicle have been proposed. This study was performed to measure antibacterial activity and continuous local antibiotic release from different concentrations of vancomycin-impregnated fibrin sealant in vitro. Methods Antibacterial activity was investigated by disk diffusion test by measuring the diameter of the growth inhibition zone of bacteria (methicillin-resistant Staphylococcus aureus, ATCC29213) from vancomycin-embedded fibrin sealant disc diluted at five different concentrations (C1-C5; 8.33, 4.167, 0.83, 0.083, and 0.0083 mg/disc, respectively). Continuous and conditioned release of vancomycin concentration (for 2 weeks and for 5 days, respectively) were also measured using high-performance liquid chromatography (HPLC) method. To mimic the physiologic wound conditions with in vitro, conditioned vancomycin release in phosphate buffer solution (PBS) was measured and replaced PBS for five consecutive days, half a day or completely daily. Results In the disk diffusion test, the mean diameters of bacterial inhibition zone were 2.54±0.07 cm, 2.61±0.12 cm, and 2.13±0.15 cm (C1, C2, and C3 respectively) but 1.67±0.06 cm and 1.23±0.15 cm in C4 and C5, respectively. Continuous elution test elicited the peak release of vancomycin from the fibrin sealant at 48 hours, with continued release until 2 weeks. However, conditioned vancomycin release decreased to half or more on day 2, however, the sustainable release was measured over the therapeutic dose (10-20 μg/mL) for 5 days and 4 days in assays of half and total exchange of PBS. Conclusion This study suggests that fibrin sealant can provide an efficient vehicle for antibiotic drug release in a wide range of neurosurgical procedures and the safe and effective therapeutic dose will be at the concentration embedded of 4.167 mg/disc or more of vancomycin.

Antibiotic resistance is a serious threat that occurs globally in the health sector due to increased consumption of inappropriate antibiotics. Guidelines for prescribing antibiotics for ARTIs have been issued in general practice to promote rational antibiotic prescribing. This study was conducted to compare the effectiveness of cefixime and tetracycline as a solution to improve monitoring of appropriate antibiotic use in the treatment of ARTIs. All stock isolates were rejuvenated first, and cultured on standard media and Kirby–Bauer disc diffusion method was used for susceptibility testing in accordance with the Clinical and Laboratory Standard Institute’s (CLSI) recommendations. Identification of bacteria from a single isolate was carried out to determine which bacteria were resistant to cefixime and tetracycline. A total of 466 single isolates of bacteria were analyzed, which showed a percentage of resistance to cefixime 38.0%, and tetracycline 92.86%. Bacterial isolates were resistant to cefixime and tetracycilne was a genus of Haemophilus, Streptococcus, Corynebacterium, Staphylococcus, and bordetella. Cefixime compared to tetracycline was proven to be superior in terms of the effectiveness of ARIs treatment.

The influence of inoculum preparation in EUCAST broth dilution and Etest to detect the coexistence of resistant and susceptible Candida subpopulations (defined as polyresistance [PR]) was evaluated. Cocultures of two echinocandin-resistant and susceptible clinical C. glabrata strains were used to simulate the occurrence of mixed populations in clinical samples, and antifungal susceptibility testing was performed with standard and modified approaches of inoculum preparation.

Fosfomycin is a phosphonic acid derivative active against a wide spectrum of Gram-positive and Gram-negative pathogens. It is used for the treatment of uncomplicated urinary tract infections (uUTI) or severe infections by oral or intravenous (i.v.) administration. In order to improve its performance and robustness, the fosfomycin strip, an antibiotic gradient diffusion strip, was redeveloped and evaluated in the multicenter study summarized in this paper. ETEST fosfomycin (ETEST FO) clinical performance was evaluated by three study sites on 152 Enterococcus faecalis, 100 Staphylococcus spp. and 330 Enterobacterales in comparison with the CLSI and EUCAST agar dilution reference method. Referring to FDA performance criteria, the ETEST FO achieved 91.0% of essential (EA) and 99.0% of categorical agreement (CA) for Escherichia coli. In addition, 98.0% EA and 93.4% CA were achieved for E. faecalis, with no very major errors (VME) or major errors (ME). According to EUCAST breakpoints for intravenous fosfomycin use, Enterobacterales and Staphylococcus spp. also met ISO acceptance criteria for EA and CA (EA 91.5%, 94.0%, respectively, and CA 98.0% for both). A VME rate of 8.8% was observed for Enterobacterales but the MICs were within EA. A trend to predict lower MICs for Citrobacter spp., E. coli and Salmonella enterica and to predict higher MICs for Klebsiella pneumoniae MICs was observed, while ETEST FO should not be used for Enterobacter cloacae, because of low EA and a high VME rate. The study results support the efficiency of the novel ETEST FO, making it an easy-to-handle tool as a substitute to the classical agar dilution method.

Background: Considering the interesting role in the peptidoglycan biosynthesis pathway, the enzyme UDP-N-acetylglucosamine enolpyruvyl transferase is an attractive target to develop new antibacterial agents. It catalyzes the first key step of this pathway and its inhibition leads to bacterial cell death. Fosfomycin is known as the natural inhibitor of MurA. Objective: The study aimed to introduce new inhibitors of MurA by virtual screening of different chemical compounds libraries, and test the best scored “virtual hits” against three pathogenic bacteria: Escherichia coli, Bacillus subtilis and Staphylococcus aureus. Methods: A virtual screening of the structural analogues of fosfomycin downloaded from the Pub- Chem database was performed. Moreover, French National Chemical Library and ZINC database were also utilized to identify new structures different from fosfomycin. FlexX was the software used for this study. The antibacterial testing was divided into two methods: disk diffusion and broth dilution. Results: A set of virtual hits was found to have better energy score than that of fosfomycin, seven of them were tested in vitro. In addition, the disk diffusion method explored four compounds that exhibited antibacterial activity: CID-21680357 (fosfomycin analogue), AB-00005001, ZINC04658565, and ZINC901335. The testing was continued by broth dilution method for both compounds CID-21680357 and ZINC901335 to determine their minimum inhibitory concentrations, and ZINC901335 had the best value with 457μg/ml against Staphylococcus aureus. Conclusion: Four compounds were found and proven in silico and in vitro to have antibacterial activity: CID-21680357, AB-00005001, ZINC04658565, and ZINC901335.

Background/Purpose The aim of this study was to characterize the S. aureus strains isolated from periodontal lesions of patients, to determine the expression of genes involved in cell adhesion upon their infection of human epithelial cells using an in vitro model, its biofilm formation, and its resistance to antibiotics. Methods S. aureus was analysed by PCR, Kirby-Bauer, and pulsed-field gel electrophoresis (PFGE), measuring gene expression by real-time PCR after infection of human cells in vitro . Results S. aureus was identified in 18.6% (50/268) of the samples. All strains (n = 50) possessed the virulence genes spa ( Staphylococcal protein A) , coa (coagulase), and icaAB (intercellular adhesin); 96% (n = 48) possessed clfB (clumping factor B), and 88% (n = 44) possessed ebps (elastin-binding protein) and sdrD (serine aspartate repeat protein D). All strains were resistant to methicillin, ampicillin, dicloxacillin, cefotaxime, and penicillin, and were multidrug resistant to 6–12 antibiotics. PFGE analysis showed 37 different pulsed-field types and most strains (60.4%) had a unique pulsed-field type. Twenty-four distinct combinations of virulence genes and antibiotic-resistant phenotypes were identified. Conclusion Although S. aureus has been considered a transient member of the oral microbiota, our results indicate a high-level expression of virulence genes and multidrug resistance in the strains isolated from periodontal lesions. These strains might complicate the successful treatment of the disease.

Disk diffusion is a slow but reliable standard method for measuring the antimicrobial susceptibility of microorganisms. Our objective was to improve the turnaround time for this method by reducing the time that cultures are incubated before setting up disk diffusion testing. For initial method development, clinical isolates (n = 13) and quality control strains (n = 8) of bacteria were inoculated on blood agar and were incubated at 35°C for either 6, 10, or 24 h before performing disk diffusion testing, in triplicate, using a panel of clinically appropriate antimicrobial agents. Disk diffusion zone sizes were interpreted using Clinical and Laboratory Standards Institute (CLSI) guidelines. Compared to standard 24 h of incubation, early 6-h growth had 1.3% major errors (MEs) and 1.9% very major errors (VMEs), whereas 10-h growth yielded 0.7% MEs and no VMEs. Categorical agreement with standard incubation was similar for both 6 h (96.7%) and 10 h (96.7%) growth. Inhibitory zone size from 6 h (r(2) = 0.98) and 10 h (r(2) = 0.99) growth correlated well with results from standard conditions. Based on these results, we performed disk diffusion under optimized conditions (6 h growth), using 100 additional clinical isolates, demonstrating a high level of categorical agreement (917 of 950 measurements [96.5%]; 95% confidence interval [CI], 95.2 to 97.5%), as well as no VMEs or MEs. Using early growth for disk diffusion testing is a simple and accurate method for susceptibility testing that can reduce time to results by as much as 18 h, compared to standard incubation, with no additional supply costs or equipment/instrumentation.

Helicobacter pylori is an important human pathogen associated with peptic ulcer disease, dyspepsia, and gastric malignancy. Antimicrobial susceptibility testing (AST) is often requested for patients who fail eradication therapy. The Clinical and Laboratory Standards Institute (CLSI) reference method, agar dilution (AD), is not performed in most laboratories and maintaining organism viability during transit to a reference laboratory is difficult. We assessed the performance of the Etest (bioMérieux) as a method for H. pylori AST in comparison to AD. Etest MICs were determined for 83 H. pylori isolates at ARUP and Cleveland Clinic (CC). Categorical agreement (CA), very major, major, and minor errors (VME, ME, and mE) were determined for Etest using AD performed at Mayo Clinic Laboratories as the reference method. Testing on isolates with errors was repeated to determine final results summarized below. For clarithromycin, 66.3% of isolates were resistant (R) by AD; Etest results at each laboratory showed 1mE (1.2%) and 1 ME (3.8%). For tetracycline, only 2 isolates were R by AD; a single VME occurred at both sites (98.8% CA, 50% VME) with the same isolate. Applying EUCAST levofloxacin breakpoints to interpret ciprofloxacin results, 60.2% of isolates were R by AD; ARUP CA was 97.6% (1 ME (3%), 1 VME (2%)) and CC CA was 96.3% (1 ME (3%), 2 VMEs (4%)). Despite high error rates, the categorical agreement was acceptable (90%) for all three antibiotics between AD and Etest. In-house susceptibility testing by gradient diffusion can allow for testing of fastidious organisms that may not survive transport to specialized laboratories; however, the method is not without technical challenges. Characterization of resistance mechanisms, increased AD dilutions, and testing from the same inoculum may determine if the observed errors reflect technical issues or breakpoints that need optimization.

Introduction: The dramatic increase in the prevalence and clinical impact of infections caused by Carbapenemase-Producing Bacteria in the nosocomial setting in Latin America represents an emerging challenge to public health. The present study detected carbapenemase-producing Gram-negative bacteria in patients from a Hospital from Venezuela, by phenotypic and genotypic methods. Methodology: The bacterial identification was carried out using conventional methods. The resistance to carbapenems was performed by Kirby-Baüer disk diffusion method, according to CLSI recommendations. The modified Hodge Test, double-disk with phenylboronic acid, double-disk with EDTA and Blue Carba Test were performed to detect phenotypic carbapenemase producers. The carbapenemase-encoding genes blaKPC, blaVIM, blaIMP, blaOXA-2, blaOXA-3, blaOXA-15 and blaOXA-21 were determined. Results: The bacterial species identified were Klebsiella pneumoniae complex (181), Pseudomonas aeruginosa (51), and Acinetobacter baumannii-calcoaceticus complex (119). KPC-type was detected in 40.17% of isolates and VIM-type in 14.53%. KPC-type gene was only identified in K. pneumoniae isolates (77.9%). VIM-type gene was identified in P. aeruginosa (86.27%) and K. pneumoniae isolates (3.87%). There was not detection of IMP-type and OXA-type genes. Conclusions: We found a predominance of K. pneumoniae KPC producers and a high rate of VIM-producing P. aeruginosa. The epidemiology of CPB in Venezuela is rapidly evolving, and enhanced surveillance and reporting are needed across the healthcare continuum.

BACKGROUND AND OBJECTIVE Jatropha variegata is traditionally used in Yemeni folk medicine for antiseptic and hemostatic purposes. In this study, the methanolic extract of the plant leaves was evaluated for its antioxidant, antibacterial and wound healing activity. MATERIALS AND METHODS The antioxidant activity was assessed using 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay. The antibacterial activity against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa was tested using disc diffusion and broth microdilution assays. In vivo, the ability of the extract to accelerate wound healing in rats was evaluated using both wound area measurements and histological analyses. RESULTS The leaves extract exhibited strong antioxidant activity with an IC50 value of 16.7 μg mL-1. The extract exhibited moderate antibacterial activity against S. aureus with inhibition zones of 10.6 mm, and the Minimum Inhibitory Concentration (MIC) value was 5 mg mL-1. The extract significantly accelerated the rate of wound healing closure compared to those treated with the vehicle. In addition, histopathological analyses of wound granulation tissues showed significantly better healing signs after 14 days in the extract-treated groups, with denser collagen deposition at the injury site. CONCLUSION The leaves of J. variegata appear to contain bioactive compounds that may be utilized clinically in combating oxidative stress and in wound management.

BACKGROUND AND OBJECTIVE Commiphora gileadensis is a plant in the Burseraceae family that grows in the western area of Saudi Arabia. Traditionally, it is used in the treatment of some superficial infections. MATERIALS AND METHODS The methanolic extract of Commiphora gileadensis isolated from its leaves and branches. The in vitro study was conducted to determine the effect of this extract on Methicillin-Resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa using an agar diffusion and Minimum inhibitory concentration (MIC) methods. The in vivo study was conducted through two different methods. The first method, 20 male Balb c-1 mice were used for the determination of Commiphora gileadensis methanolic extract toxicity (LD50). In the second method, 40 male mice were used and were put into four groups. The first and second groups were injected subcutaneously with 108 CFU of MRSA 1 mL-1, while the third and fourth groups were injected with 108 CFU of Pseudomonas aeruginosa 1 mL-1. The comparison between groups was done by using a t-test (p

Contamination of ready-to-eat (RTE) foods by pathogenic bacteria may predispose consumers to foodborne diseases. This study investigated the presence of bacterial contaminants and their antibiotic susceptibility patterns in three locally processed RTE foods (eko, fufu and zobo) vended in urban markets in Ogun state, Nigeria. Bacteria isolated from a total of 120 RTE food samples were identified by 16S rRNA gene phylogeny while susceptibility patterns to eight classes of antibiotics were determined by the disc diffusion method. Species belonging to the genera Acinetobacter and Enterobacter were recovered from all RTE food types investigated, Klebsiella and Staphylococcus were recovered from eko and fufu samples, while those of Shigella were recovered from eko samples. Enterobacter hormaechei was the most prevalent species in all three RTE food types. Precisely 99% of 149 isolates were multidrug-resistant, suggesting a high risk for RTE food handlers and consumers. Co-resistance to ampicillin and cephalothin was the most frequently observed resistance phenotype. Results demonstrate that improved hygiene practices by food processors and vendors are urgently required during RTE processing and retail. Also, adequate food safety guidelines, regulation and enforcement by relevant government agencies are needed to improve the safety of RTE foods and ensure the protection of consumer health.

ObjectivesThe emergence of multidrug-resistantNeisseria gonorrhoeae(NG) is a major global health threat necessitating response and control measures. NG antimicrobial resistance (AMR) surveillance data from sub-Saharan countries is exceedingly limited. This paper aims to describe the establishment, design and implementation of a standardised and quality-assured gonococcal surveillance programme and to describe the susceptibility patterns of the cultured gonococcal isolates in Kampala, Uganda.MethodsFrom March 2018 to September 2019, using the WHO Enhanced Gonococcal Antimicrobial Surveillance Programme (EGASP) protocol, consecutive males with urethral discharge syndrome were recruited from 10 surveillance sites in Kampala City, Uganda, in collaboration with the Ministry of Health. Males completed a questionnaire and provided a urethral swab specimen. Culture, identification and antimicrobial susceptibility testing (Etest) were performed.ResultsOf the 1013 males recruited, 73.1% (740/1013) had a positive Gram stain and 51.1% (n=518) were culture-positive for NG. Using Etest (458 isolates), the resistance to ciprofloxacin was 99.6%. Most isolates were susceptible to azithromycin, cefoxitin and gentamicin, that is, 99.8%, 98.5% and 92.4%, respectively, and all isolates were susceptible to ceftriaxone and cefixime.ConclusionsWe established a standardised, quality-assured WHO EGASP. Using Etest, 458 isolates were characterised, with associated epidemiological surveillance data, in 1.5 years, which by far exceed the minimum 100 isolates per year and country requested in the WHO Global GASP, to detect AMR levels with confidence. These isolates with the epidemiological data can be used to develop population level interventions.

Objectives The aim of this study was to evaluate antibiotic susceptibility patterns in commercially available dietary and probiotic supplements. Methods Probiotic strains were isolated from the dietary supplements (designated as sample B, D and V), and multidrug resistance profiles were tested using the Kirby-Bauer test. Minimum inhibitory concentrations and double-disk synergy tests were performed to detect the mechanism of action of the resistance, and the presence of extended spectrum β-lactamase activity (ESBL) was confirmed. Results The isolates Streptococcus faecalis and Bacillus mesentericus (both from sample B) were found to be resistant to penicillin G, Lactobacillus acidophilus (sample D) was resistant to ampicillin, and all the isolates from samples B, D and V were resistant to ceftazidime. The isolates Lactobacillus sporogenes, S. faecalis, B. mesentericus from sample B, Lactobacillus. rhamnosus, Saccharomyces boulardii from sample D, and L. sporogenes (sample V) were resistant to erythromycin. Conclusions The findings showed the presence of antibiotic resistance in probiotic bacteria isolated from commercially available dietary supplements. Because multidrug resistance is a serious emerging issue, and the risk of drug-resistant gene transfer to commensals or pathogens of the gut is inevitable, the safety of probiotics has become a major criterion of interest. The findings of this study would serve as a platform for further screening and characterization of the determinants of antibiotic resistance and the genetic mechanisms of resistance.

Bacterial soft rot caused by the bacteriaDickeyaandPectobacteriumis a destructive disease of vegetables, as well as ornamental plants. Several management options exist to help control these pathogens. Because of the limited success of these approaches, there is a need for the development of alternative methods to reduce losses. In this study, we evaluated the effect of potassium tetraborate tetrahydrate (PTB) on the growth of sixDickeyaandPectobacteriumspp. Disc diffusion assays showed thatDickeyaspp. andPectobacteriumspp. differ in their sensitivity to PTB. Spontaneous PTB-resistant mutants ofPectobacteriumwere identified and further investigation of the mechanism of PTB resistance was conducted by full genome sequencing. Point mutations in genescpdBandsupKwere found in a singlePectobacterium atrosepticumPTB-resistant mutant. Additionally, point mutations in genesprfB(synonymsupK) andprmCwere found in two independentPectobacterium brasiliensePTB-resistant mutants.prfBandprmCencode peptide chain release factor 2 and its methyltransferase, respectively. We propose the disruption of translation activity due to PTB leads toPectobacteriumgrowth inhibition. TheP. atrosepticumPTB-resistant mutant showed altered swimming motility. Disease severity was reduced forP. atrosepticum-inoculated potato stems sprayed with PTB. We discuss the potential risk of selecting for bacterial resistance to this chemical.

Aim: A sodium alginate-based biohydrogel was prepared integrating choline oleate deep euthetic solvent as facilitator of transdermal delivery and a cocktail of lytic bacteriophages for Acinetobacter baumannii, aiming at treating soft-tissue infections by the aforementioned pathogen. Materials & methods: Two bacteriophages were isolated from a hospital sewage and a wastewater treatment plant sewage in Sorocaba (Brazil), and characterized via SDS-PAGE electrophoresis, transmission electron microscope and evaluation of lytic spectra of the bacteriophage cocktail. The biohydrogel was prepared and characterized by DSC, FTIR, XRD, DESEM, XRT and transdermal permeation of the bacteriophage cocktail. Results & conclusion: The physico-chemical characterization of the biohydrogel produce indicated adequate structural characteristics and ability to promote/facilitate transdermal delivery of bacteriophage particles, thus showing potential for biopharmaceutical applications.

Background Colistin has become a last-resort antibiotic for the management of multidrug-resistant gram-negative bacteria. The disk diffusion test is cheap and easy to perform but may be unreliable for colistin susceptibility testing due to poor diffusion of the large colistin molecule. An improved agar diffusion test would increase the reliability of colistin susceptibility testing. This study aimed to modify Muller-Hinton agar (MHA) to improve colistin diffusion in agar. Methods MHA was modified by reducing the agar concentration from 100% to 30% and supplementing with protamine. We tested 60 gram-negative clinical isolates of Pseudomonas aeruginosa (N=27) and Acinetobacter calcoaceticus-baumannii complex (N=33). Disk diffusion test results were interpreted based on minimum inhibitory concentrations determined by broth microdilution. Results The modified MHA yielded the best performance metrics, including 94.7% sensitivity, 100% specificity, and an area under the curve of 0.995 (95% confidence interval, 0.982–1.000), P

Methicillin-resistant Staphylococcus aureus (MRSA), an important widespread cause of severe infection in both humans and animals, is a significant pathogen of public health concern. This study examined the presence of MRSA in 400 samples comprising 200 raw milks (145 from goat and 55 from sheep) and 200 nasal swabs (145 from goats and 55 from sheep) collected from ten different locations in Abeokuta, Nigeria. Samples were examined using standard bacteriological methods for the isolation and identification of Staphylococcus aureus and culture on oxacillin (6 μg/ml) and cefoxitin (2 μg/ml) selective media for the detection of MRSA. Suspected MRSA isolates were confirmed by latex agglutination test for the detection of penicillin-binding protein 2a (PBP2a). Antibiotic susceptibility testing was determined by Kirby Bauer disc diffusion method. Staphylococcus aureus was detected in 72 (18%) of 400 samples of which 52 (13%) were confirmed as MRSA. Methicillin-resistant S. aureus was detected in raw milk (37 of 200; 18.5%) and nasal swab (15 of 200; 7.5%). There was no significance difference (p > 0.05) in the prevalence of MRSA in sheep (37.7%) and goat (23.4%). The MRSA isolates showed resistance to ampicillin (100%), cloxacillin (100%), sulphamethoxazole-trimethoprim (100%), amoxicillin-clavulanate (84.6%), ceftriaxone (75%), cefuroxime (69.2%), erythromycin (65.4%), streptomycin (38.5%), ciprofloxacin (23.1%), pefloxacin (21.2%) and gentamicin (17.3%). The presence of multidrug-resistant MRSA in small ruminants reared in Abeokuta metropolis may be due to regular use of antibiotics and unhygienic practices by farmers. This in turn constitutes a potential public health risk to the owners, consumers of small ruminant products and the general populace.

Resumen Introducción. La aparición de enterobacterias multirresistentes y productoras de betalactamasas de espectro extendido en pacientes ambulatorios con infecciones urinarias representa un problema de salud pública en Perú. Objetivo. Comparar los perfiles de resistencia de Escherichia coli uropatógenas e identificar los fenotipos de cepas productoras de betalactamasas de espectro extendido en tres establecimientos privados de salud localizados en las regiones de la costa, la sierra y la selva de Perú. Materiales y métodos. Se llevó a cabo durante el 2016 un estudio descriptivo de 98 muestras de orina de pacientes con infección urinaria, 35 procedentes de Lima (costa), 38 de Juliaca (sierra) y 25 de Iquitos (selva), en el que se determinó la sensibilidad antimicrobiana utilizando ocho discos antibióticos. Asimismo, se evaluó la producción de betalactamasas de espectro extendido con discos de cefotaxima, de ceftazidima o de su combinación, con ácido clavulánico en agar Mueller-Hinton. Resultados. Se identificaron 18 perfiles de resistencia que incluían desde los sensibles a todos los antibióticos hasta los resistentes simultáneamente a siete antibióticos, con el 18,4 % de aislamientos resistentes a un antibiótico y el 54,0 % de multirresistentes. Se detectó producción de betalactamasas en el 28,6 % de las cepas procedentes de la región de Puno. También, se observó un mayor número de casos en el rango de edad de 31 a 45 años con resistencia a ceftazidima, ceftriaxona, gentamicina y trimetoprim-sulfametoxazol en el establecimiento de salud de Puno. Conclusión. Los perfiles de resistencia variaron según la localización geográfica del establecimiento de salud, observándose mayor resistencia a los antibióticos en la región de la sierra de Perú, con el 28,6 % de cepas productoras de betalactamasas de espectro extendido. Abstract Introduction: The appearance of multidrug-resistant and beta-lactamase producing enterobacteria in outpatient care facilities represent a public health problem in Perú. Objective: To compare the resistance profiles of uropathogenic Escherichia coli and to identify extended-spectrum beta-lactamase-producing phenotypes in three private health facilities located in the Peruvian coast, Andean and jungle regions. Materials and methods: We conducted a descriptive study on 98 urine samples from Lima (coast), Juliaca (Andean region) and Iquitos (jungle region) during 2016. We determined the antimicrobial susceptibility in 35 samples from Lima, 38 from Juliaca and 25 from Iquitos using eight antibiotic disks in samples from patients diagnosed with urinary infection. We also evaluated the production of extended-spectrum beta-lactamases with cefotaxime and ceftazidime disks and a combination of both with clavulanic acid on Mueller-Hinton agar. Results: We identified 18 resistance profiles ranging from those sensitive to others simultaneously resistant to seven antibiotics: 18.4% resistant to one and 54.0% to multiple antibiotics. We detected beta-lactamase production in 28.6% of the strains from the Puno region. Likewise, we observed a greater number of cases with resistance to ceftazidime, ceftriaxone, gentamicin, and trimethoprim-sulfamethoxazole in Puno’s health facility in patients within the 31 to 45 year age range. Conclusion: Resistance profiles varied according to the geographical location of the health facilities under study. Resistance to antibiotics was higher in the Andean region with 28.6% of strains producing extended-spectrum beta-lactamases.

Background: Empirical antibiotherapy (EA) should target all bacteria in post-operative peritonitis (PP). Nevertheless, recent studies failed to prove a link between adequacy of EA and prognosis of PP. We sought to confirm this loss of association between adequate EA and prognosis and to analyze the evolution of patients' characteristics and antimicrobial strategies. Methods: This is was retrospective study. Patients with a positive fungal culture were excluded. The cohort was divided into two time periods. Data of survivors and non-survivors were compared within each time period. Differences between the two periods were assessed. A multivariable analysis searched for parameters associated with a higher hospital mortality rate. Results: Two hundred fifty-one patients were included, with 92 patients in the first period (P1) and 152 patients in the second period (P2). Inadequate EA was associated with a worse outcome only in P1. The multivariable analysis in the whole cohort showed that inadequate EA was associated with a higher mortality rate. When the differences noticed between the two periods were entered in the model (presence of resistant gram-positive cocci and EA comprising glycopeptides), inadequate EA was no longer associated with worse outcome. In P1, the most severe patients had more resistant bacteria, hence, had a higher rate of inadequate EA. This artifact disappeared in P2, during which broader antibiotherapies with triple EA were more often prescribed for the most severe patients. Conclusion: This study showed that the link between inadequate EA and outcome of patients with PP was at least partly artifactual in older studies.

Objectives Enterococci have gained attention during the past decade as important nosocomial pathogens. Their increasing prevalence has been paralleled by the occurrence of multidrug-resistant and high-level aminoglycoside-resistant strains. This study isolated Enterococcus spp. from hospital samples and determined their antibiotic resistance profile, focusing on aminoglycosides, and associated resistance mechanisms. Methods A total of 195 enterococci from hospital samples in Tehran were studied. Isolates were identified by biochemical reactions. Antimicrobial resistance was determined by disk diffusion. The vancomycin MIC for vancomycin-resistant isolates was determined by agar dilution. Detection of aminoglycoside resistance genes and intI1 and intI2 gene was performed by PCR. Results The majority of isolates were Enterococcus faecalis (65.1%), followed by Enterococcus faecium (31.8%), Enterococcus gallinarum (2.6%) and Enterococcus solitarius (0.5%). According to antibiogram results, 42.1% of isolates were high-level gentamicin-resistant (HLGR) and 40.5% were high-level streptomycin-resistant (HLSR). There was a high prevalence of aac(6')-Ie–aph(2")-Ia (96.3%) among HLGR isolates. ant(6)-Ia and aadA were identified in 93.7% and 64.6% of HLSR isolates, respectively. aph(2ʹʹ)-Ic was detected in 7 isolates (3.6%) and aph(2ʹʹ)-Ib in only 4 isolates (2.1%); no isolates harboured aph(2ʹʹ)-Id, intI1 or intI2. Conclusion Multidrug resistance was higher among HLGR and HLSR isolates compared with non-HLGR and non-HLSR isolates, which may result in limited treatment options. More than 50% of isolates were susceptible to aminoglycosides, thus correct identification in clinical laboratories and administration of these antibiotics can result in decreased used of antibiotics such as vancomycin and linezolid and help to reduce the emergence of resistance to these drugs.

Objectives: Due to the rapid emergence of multidrug-resistant isolates, Campylobacter jejuni (C. jejuni) has been classified as a member of the priority pathogens group. This study aimed to determine the prevalence, antibiotic resistance patterns and source tracking of clinical C. jejuni isolates from paediatric diarrhoeal patients in Pakistan. Methods: A total of 150 stool samples from children were processed for the presence of C. jejuni using culture, biochemical tests and species-specific PCR. Antibiotic susceptibility of the isolates was determined by the disc diffusion method, and metallo-β-lactamase (MBL) producers were detected using gene-specific PCR. Source tracking was performed using source-predictive PCR. Results: C. jejuni was present in 54.6% of the processed samples. More than 80% of the isolated strains were resistant to seven of 12 tested antibiotics. High levels of susceptibility were observed against imipenem (12.2%) and TGC (9.7%). Six isolates (7.3%) were MBL producers and positive for at least one of the five MBL genes. Source tracking showed that 57.3% of the isolates belonged to livestock-associated clusters (C1–C6) and 42.8% were assigned to non-livestock/environmental clusters (C7–C9). Isolates belonging to livestock clusters had a high Multiple Antibiotic Resistance (MAR) index (P < 0.001) as compared with non-livestock. Conclusion: A high prevalence of multidrug-resistant C. jejuni among paediatric diarrhoeal patients was observed. Moreover, the association of these isolates to livestock clades suggests transmission to human populations via the food chain. The presence of imipenem-resistant MBL-producing C. jejuni can lead to serious public health concerns.

International audience; Objectives: The aim of this study was to describe antibiotic prescribing patterns and antimicrobial resistance rates in hospitalised children with febrile and afebrile urinary tract infections (UTIs).Methods: Antibiotic prescriptions and antibiograms for neonates, infants and older children with UTI admitted to a general district hospital in Central Greece were evaluated. Data covering a 5-year period were collected retrospectively from the Paediatric Department's Electronic Clinical Archive. Patients were included based on clinical and microbiological criteria. Antimicrobial susceptibility was determined by the Kirby-Bauer disk diffusion method.Results: A total of 230 patients were included in the study. Among 459 prescriptions identified, amikacin (31.2%) was the most common antibiotic prescribed in this population, followed by amoxicillin/clavulanic acid (17.4%) and ampicillin (13.5%). Children received prolonged intravenous (i.v.) treatments for febrile (mean ± S.D., 5.4 ± 1.45 days) and afebrile UTIs (mean ± S.D., 4.4 ± 1.64 days). A total of 236 pathogens were isolated. The main causative organism was Escherichia coli (79.2%) with high reported resistance rates to ampicillin (42.0%), trimethoprim/sulfamethoxazole (26.5%) and amoxicillin/clavulanic acid (12.2%); lower resistance rates were identified for third-generation cephalosporins (1.7%), nitrofurantoin (2.3%), ciprofloxacin (1.4%) and amikacin (0.9%). Klebsiella spp. isolates were highly resistant to cefaclor (27.3%).Conclusion: High prescribing rates for amikacin and penicillins (± β-lactamase inhibitors) and prolonged i.v. treatments were observed. Escherichia coli was highly resistant to ampicillin, whilst third-generation cephalosporins exhibited greater in vitro efficacy. Establishment of antimicrobial stewardship programmes and regular monitoring of antimicrobial resistance could help to minimise inappropriate prescribing for UTIs.

Objectives This study evaluated disc diffusion tests and agar screening for detecting mecA-mediated oxacillin resistance in Staphylococcus lugdunensis (S. lugdunensis). Methods Staphylococcus lugdunensis isolates (n = 179) from diverse sources in Hong Kong during 1998–2018 were investigated by disc diffusion tests (cefoxitin and oxacillin) and inoculation onto oxacillin (1 μg/mL and 2 μg/mL) and chromID methicillin-resistant Staphylococcus aureus (MRSA) agars. The results were compared with mecA PCR as the reference. Isolates with discordant results were further tested by MIC and penicillin-binding protein 2a (PBP2a) assays. Results Cefoxitin and oxacillin zone diameters were not distributed in ways that allowed reliable division of the mecA-positive (n = 52) and mecA-negative (n = 127) isolates. On applying the 2019 Clinical Laboratory Standards Institute (CLSI) M100 breakpoints for cefoxitin disc results, there was 88% categorical agreement (CA) and 40% very major error (VME). Screening using 2 μg/mL oxacillin agar reliably differentiated mecA-positive and mecA-negative isolates (100% CA) without any major error (ME) or VME results. The performance of screening using 1 μg/mL oxacillin agar or ChromID MRSA agar was variable (74–89% CA, 0–38% ME and 0–37% VME). The mecA-positive isolates (n = 21) that could not be detected by the cefoxitin disc test were further characterised. The cefoxitin MIC for all 21 isolates was ≤4 μg/mL. Twenty isolates had an oxacillin MIC of 1–2 μg/mL and one had an oxacillin MIC of 4 μg/mL. All had positive PBP2a results and were typed as clonal cluster 27/SCCmec V. Conclusions These findings highlight the need to evaluate phenotypic methods using mecA-positive S. lugdunensis with different oxacillin resistance phenotypes.

Introduction: The use of antimicrobial discs remains one of the main methods for assessing antibiotic activity. Most discs are made to one of three main standards (FDA, WHO, DIN); these all describe an assay method for assessing the quality of discs using a linear method. Theory predicts a curved relationship, and this is backed up in many cases in practice. In such cases, the assays are potentially invalid. Other sources of uncertainty arising from the manufacturing processes employed are also discussed.Areas covered: This includes error arising from applying FDA, WHO, and DIN standards, the manufacturing techniques employed, and variation in the materials used in production. The need for a specification that relates directly to the use of the discs is also discussed.Expert opinion: Manufactured discs, some of which may be out of specification due to curvature and other sources of error, have been used to establish quality zone sizes. Quality zone sizes have then been used to measure the quality of discs. This circular quality system where there is no quantitative check is potentially unsafe. In the many decades of their use, there has been no comprehensive check on the quality of manufactured discs using quantitative, validated assays.

Objectives: Levonadifloxacin is a novel benzoquinolizine subclass of quinolone with broad-spectrum activities against problematic pathogens such as methicillin-resistant Staphylococcus aureus, quinolone-resistant S. aureus, vancomycin intermediate S. aureus, and vancomycin-resistant S. aureus. Levonadifloxacin and its oral prodrug, alalevonadifloxacin, have been recently approved in India for the treatment of acute bacterial skin and skin structure infections, including concurrent bacteraemia and diabetic foot infections. The aim of the study is to assess the activity of levonadifloxacin against Gram-positive clinical isolates collected from various Indian hospitals using the disc-diffusion method. Materials and Methods: Nonduplicate isolates of S. aureus and other Gram-positive isolates collected from June 2019 to March 2020 were subjected to levonadifloxacin susceptibility testing (disk diffusion method) as per the Clinical and Laboratory Standards Institute guidelines (Year 2019). Levonadifloxacin 10 μg impregnated disks were used during the testing. Results: A total of 664 diverse Gram-positive clinical isolates collected from six different hospitals in India were analyzed. Majority (65.5%) of the isolates were S. aureus. All the S. aureus and other Gram-positive isolates were found to be susceptible to levonadifloxacin as per the prespecified interpretive criteria identified based on population pharmacokinetic model and Monte Carlo simulation enabled probability of pharmacodynamic target attainment analysis. Conclusions: The present study showed that levonadifloxacin was highly active against contemporary Gram-positive pathogens and furthermore demonstrated that levonadifloxacin susceptibilities can be reliably determined using the disc-diffusion method.

We analyzed the potential antibacterial effects of two different PdB against methicillin-resistant S. aureus and P. aeruginosa. The third-degree burn wound healing effects of PdB was also studied. Blood samples were obtained from 10 healthy volunteers and biological assays of the PdB were performed and the antimicrobial activity against MRSA and P. aeruginosa was determined using disk diffusion (DD), broth microdilution (BMD), and time-kill assay methods. 48 Wistar albino rats were burned and infected with MRSA. Two groups were injected PdB, the control groups were treated with plasma and received no treatment respectively. In the next step, the rats were euthanized and skin biopsies were collected and histopathologic changes were examined. The results of DD and BMD showed that both PdB performed very well on MRSA, whereas P. aeruginosa was only inhibited by F-PdB and was less susceptible than MRSA to PdBs. The time-kill assay also showed that F-PdB has an antibacterial effect at 4 hours for two strains. Histopathological studies showed that the treated groups had less inflammatory cells and necrotic tissues. Our data suggest that PdB may possess a clinical utility as a novel topical antimicrobial and wound healing agent for infected burn wounds.

Background The existing modified carbapenem inactivation methods (mCIMs) recommended by the CLSI for detecting carbapenemase production have not been applicable for Acinetobacter baumannii. We evaluated the influence of matrices used in mCIMs and CIMTris on the stability of the disks for detecting carbapenemase producers and suggested optimal mCIM conditions for detecting carbapenemase-producing A. baumannii. Methods Seventy-three A. baumannii isolates characterized for antimicrobial susceptibility and carbapenemase encoding genes were tested for carbapenemase production using mCIM and CIMTris. The influence of the matrices (Tryptic soy broth [TSB] and Tris-HCl) used in these methods on the stability of the meropenem (MEM) disk was also evaluated. The mCIM conditions were adjusted to enhance screening sensitivity and specificity for detecting carbapenemase-producing A. baumannii. Results The matrices had an impact on the stability of the MEM disk after the incubation period (two or four hrs). TSB nutrient broth is an appropriate matrix for mCIM compared with Tris-HCl pH 7.6, which leads to the loss of MEM activity in CIMTris. The sensitivity and the specificity of the optimal mCIM were both 100%. Conclusions We established optimal mCIM conditions for simple, accurate, and reproducible detection of carbapenemase-producing A. baumannii.

Purpose: Emergence of multidrug-resistant and methicillin-resistant Staphylococcus aureus (MRSA) infections in HIV patients limit the treatment options and challenge the clinical management of infections. The periodic monitoring of S. aureus infections and its drug resistance profile in HIV patients are of paramount importance in clinical management. Materials and methods: A total of 7204 clinical specimens from HIV patients from 2012 to 2017 were processed for the isolation of S. aureus strains using conventional culture techniques and cultures were identified using standard biochemical test. Antibiotic susceptibility of S. aureus strains was tested by Kirby-Bauer disk diffusion method. Results: A total of 380 (5.3%) S. aureus strains were isolated from HIV patients in the study period. High percentage of S. aureus strains were isolates from urine (69.5%) specimen and 58.4% of S. aureus infections were noted among hospitalized patients. Antibiotic susceptibility profile reveals S. aureus was highly resistant to penicillin (95.2%) followed by cephalexin (84.6%). Methicillin resistance was highly observed in the year 2017 (86%) and the rate of MRSA steadily increasing from 51.8% in 2012 to 86% in 2017. Significant increase of S. aureus infections (35%; p

The aim of this laboratory-based study was to compare carbapenem MICs yielded by Sensititre, Vitek 2, MicroScan WalkAway plus and Etest for Oxacillin (OXA)-48-like Klebsiella pneumoniae isolates.Analysis was performed for categorical agreement for ertapenem, meropenem, and imipenem, and the proportion of isolates with MICs ≤8μg/mL and the MIC50/MIC90 for meropenem and imipenem, from a convenience sample of 82 deduplicated blood culture OXA-48-like K. pneumoniae isolates.The proportion of isolates testing susceptible to ertapenem by Etest (19/82, 23.1%) differed from Sensititre/Vitek (0/82) and MicroScan (2/82, 2.4%) (p 0.001 for all). For meropenem, the proportion of isolates susceptible by Etest (31/82, 37.8%) differed from Sensititre/Vitek (16/82, 19.5%) (p = 0.015). There was variation in the proportion of isolates that tested imipenem susceptible when comparing Sensititre (9/82, 11%) and Vitek (8/82, 9.8%) to MicroScan (27/82, 32.9%), p = 0.001 and p 0.001, respectively, Sensititre and Vitek to Etest (45/82, 54.9%), p 0.001 for both, and MicroScan to Etest, p = 0.007. The proportion of isolates with meropenem MICs ≤8μg/mL with Sensititre and Vitek differed significantly from Etest, 58.5% and 85.4%, respectively, p 0.001. A 2-fold difference between the Sensititre and Vitek meropenem and imipenem MIC at which ≥50% of isolates were inhibited compared to the MicroScan, and a 4-fold difference compared to Etest, was present.Substantial variability in carbapenem MICs for OXA-48-like carbapenemase-producing Enterobacterales isolates by the four methods was demonstrated. Performance characteristics verification of MIC methods in use for the predominant carbapenemase-producing Enterobacterales type is required by laboratories to optimize the accuracy of carbapenem reporting.

Plazomicin (PLZ), brand name ZEMDRI (Cipla Therapeutics), is a novel aminoglycoside antibiotic approved by the US Food and Drug Administration (FDA) for treatment of complicated urinary tract infections including pyelonephritis. ETEST® is a gradient diffusion method that represents an alternative to the more laborious broth micro-dilution (BMD) method for performing antimicrobial susceptibility testing (AST). A multi-center evaluation of the performance of the new ETEST® PLZ (bioMerieux) was conducted in comparison with BMD following FDA and International Standards Organization (ISO) recommendations using FDA-defined breakpoints. Clinical isolates of Enterobacterales (n=598) were included. Fifty-three isolates were resistant to PLZ according to BMD. Overall, the ETEST® PLZ demonstrated 99.0% Essential Agreement (EA), 92.8% Category Agreement (CA), 1.9% Very Major Errors (VME), 0% Major Errors (ME) and 7.0% minor Errors (mE) with both clinical and challenge isolates of Enterobacterales. The VME was found for a single Serratia marcescens strain. Individual species demonstrated EA rates ≥ 90%. In conclusion, we report that ETEST® PLZ represents an accurate tool for performing PLZ AST of Enterobacterales.

In this cross-sectional study, the isolation and identification of Methicillin Resistant Staphylococcus aureus (MRSA) and Methicillin Resistant S. epidermidis (MRSE) was described from skin infections (n=100). Initial isolation was done by conventional procedures followed by amplification/ sequence analysis of 16S rRNA. Methicillin resistance was determined using cefoxitin discs and resistant isolates were screened for mec-A gene followed by Minimum Inhibitory Concentrations (MIC) determination of vancomycin. In second phase, we investigated extract of Azadirachta indica leaves using Fourier Transformed Infrared Spectroscopy (FTIR-Spectroscopy) and investigated in vitro activity. Initially, total of 28 Staphylococci were identified. 16S rRNA gene sequence confirmed S. aureus (22), S. epidermidis (3) and S. saprophyticus (3) isolates. Cefoxitin discs showed (7/22) MRSA, (3/3) (MRSE) and none of the methicillin resistant S. saprophyticus. MRSA and MRSE isolates showed presence of mec-A gene. However, all isolates were sensitive to vancomycin MIC (0.5-2μg/mL) and sensitive to Linezolid. FTIR-Spectroscopy of A. indica indicated the presence of azadirachtin and nimbolinin. The mean zone of inhibition was measured 14.23±1.37 and 13.66±0.70 against MRSA and MRSE isolates, respectively. Altogether, MRSA and MRSE is significant public health concern. However, vancomycin and linezolid were found effective and extract of A. indica showed in vitro effects.

This study aimed to develop a plasticized starch (PS) based film loaded with chitosan nanoparticles (CNPs, 1, 2, 3, and 4%) as a reinforcing and antibacterial agent. We examined the morphology, biodegradability, mechanical, thermo-mechanical, and barrier properties of the PS/CNPs films. The antimicrobial activity against both Gram-positive (S. aureus) and Gram-negative (E. coli) bacteria was investigated by colony forming unit (CFU) and disc diffusion methods. A dense structure was obtained for all PS/CNPs films and, thus, their complete biodegradation occurred in more days than neat PS. The increase in the CNPs percentage led to improved mechanical behaviour and barrier properties. PS-CNPs composite films revealed inhibition zones against both E. coli and S. aureus, with the 100% reduction in CFU against S. aureus. The current study exhibited that PS-CNPs films were more effective in inhibiting bacteria growth than neat PS film, confirming the composite films potential application as antimicrobial food packaging.

The objective of this research was to evaluate the correlation between inhibitory zones and MIC when testing ceftazidime-avibactam using disk diffusion, Etest, and broth microdilution method established by the Clinical and Laboratory Standards Institute (CLSI). Four-hundred and 58 isolates of

Antimicrobial resistance mediated by extended-spectrum beta-lactamases (ESBL), AmpC beta-lactamase and metallo-beta-lactamase (MBL) producing Acinetobacter species is an emerging problem worldwide. In this cross-sectional study total 341 specimens were collected over a period of one year from January 2017 to January 2018. Specimens were collected from ICU and Surgery unit of Mymensingh Medical College Hospital, Mymensingh, Bangladesh. Specimens were collected from ICU and Surgery Unit of Mymensingh Medical College Hospital, Mymensingh, Bangladesh. Samples were processed for culture by standard conventional methods and susceptibility testing and determined by Kirby Bauer disc diffusion method. Antibiotic discs and their strength were according to the CLSI 2017 guideline. Molecular study was done to detect the species by OXA-51 gene and drug resistance genes (IMP, VIM, NDM, TEM, SHV, CTX, SPM, SIM and GIM). Species identification was done by OXA-51 gene which is intrinsic to Acinetobacter baumannii. Among the 46 isolates, 36(78.26%) were positive for Oxa-51 gene, 16(34.8%) for TEM gene, 9(19.6%) for VIM gene, 3(6.5%) for NDM gene and 1(2.2%) for IMP gene. This study gives an alarming sign towards high prevalence of cephalosporin and carbapenem resistance due to production of extended spectrum beta-lactamases and metallo-betalactamases, respectively. Early detection, proper antibiotic policies, and compliance towards infection control practices are the best defenses against these organisms.

Increase in bacterial resistance to commonly used antibiotics is a major public health concern generating interest in novel antibacterial treatments. Aim of this scientific endeavor was to find an alternative efficient antibacterial agent from non-conventional plant source for human health applications. We used an eco-friendly approach for phyto-fabrication of silver nanoparticles (AgNPs) by utilizing logging residue from timber trees Gmelina arborea (GA). GC–MS analysis of leaves, barks, flowers, fruits, and roots was conducted to determine the bioactive compounds. Biosynthesis, morphological and structural characterization of GA-AgNPs were undertaken by UV–Vis spectroscopy, scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDX), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR) and X-ray diffractometer (XRD). GA-AgNPs were evaluated for antibacterial, antibiofilm, antioxidant, wound healing properties and their toxicity studies were carried out. Results identified the presence of terpenoids, sterols, aliphatic alcohols, aldehydes, and flavonoids in leaves, making leaf extract the ideal choice for phyto-fabrication of silver nanoparticles. The synthesis of GA-AgNPs was confirmed by dark brown colored colloidal solution and spectral absorption peak at 420 nm. Spherical, uniformly dispersed, crystalline GA-AgNPs were 34–40 nm in diameter and stable in solutions at room temperature. Functional groups attributed to the presence of flavonoids, terpenoids, and phenols that acted as reducing and capping agents. Antibacterial potency was confirmed against pathogenic bacteria Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus by disc diffusion assay, MIC and MBC assay, biofilm inhibition assay, electron-microscopy, cell staining and colony counting techniques. The results from zone of inhibition, number of ruptured cells and dead-cell-count analysis confirmed that GA-AgNPs were more effective than GA-extract and their bacteria inhibition activity level increased further when loaded on hydrogel as GA-AgNPs-PF127, making it a novel distinguishing feature. Antioxidant activity was confirmed by the free radical scavenging assays (DPPH and ABTS). Wound healing potential was confirmed by cell scratch assay in human dermal fibroblast cell lines. Cell-proliferation study in human chang liver cell lines and optical microscopic observations confirmed non-toxicity of GA-AgNPs at low doses. Our study concluded that biosynthesized GA-AgNPs had enhanced antibacterial, antibiofilm, antioxidant, and wound healing properties.

© 2021 Elsevier B.V.This work addresses the mathematical model building to detect the diameter of the inhibition zone of gilaburu (Viburnum opulus L.) extract against eight different Fusarium strains isolated from diseased potato tubers. Gilaburu extracts were obtained with acetone, ethanol or methanol. The isolated Fusarium strains were: F. solani, F. oxysporum, F. sambucinum, F. graminearum, F. coeruleum, F. sulphureum, F. auneaceum and F. culmorum. In general, it was observed that ethanolic extracts showed highest antifungal activity. The antifungal activity of extracts was evaluated with machine learning (ML) methods. Several ML methods (classification and regression trees (CART), support vector machines (SVM), k-Nearest Neighbors (k−NN), artificial neural network (ANN), ensemble algorithms (EA), AdaBoost (AB) algorithm, gradient boosting (GBM) algorithm, random forests (RF) bagging algorithm and extra trees (ET)) were applied and compared for modeling fungal growth. From this research, it is clear that ML methods have the lowest error level. As a result, ML methods are reliable, fast, and cheap tools for predicting the antifungal activity of gilaburu extracts. These encouraging results will attract more research efforts to implement ML into the field of food microbiology instead of traditional methods.

Trianthema triquetra Rottl. ex Willed is being used as herbal remedy for chronic ulcer, wound healing, diabetes, skin and inflammatory diseases in India and Pakistan. Still, no scientific data is available about the therapeutic potential and phytochemistry of the plant. The aim of the current investigation is to perform GC-MS analysis, antioxidant (total phenolic and flavonoid content, DPPH assay), antimicrobial (disc diffusion assay) and cytotoxic (XTT and RBC's cellular membrane protection assay) studies. Whole plant material was dried and extracted with methanol to get crude methanolic extract and then it was fractionated with n-hexane, ethyl-acetate, chloroform, n-butanol and water. Results showed that n-butanol fraction exhibited a significant (p0.05) antioxidant potential measured by DPPH assay (IC50=63.35±0.13μg/mL) and also possess highest phenolic content (177±4.36mg/g GAE). Whereas, n-hexane fraction showed highest flavonoid content (14.67±1.53mg/g QE). Two major components (2, 4-Ditert-butyl-6-nitrophenol (26.79%) and Squalene (25.64%) were detected in GC-MS analysis of chloroform fraction, eluted from column chromatography. Moreover, chloroform fraction also exhibited antibacterial activity towards all the tested strains of bacteria and fungi. Significant (p0.05) dose dependent inhibition response on cell growth against CCRF-CEM cell lines was exhibited by methanolic extract. Furthermore, hemolytic potential of methanolic extract was found to be in safe range (2.23%-6.37%). So, it can be inferred that Trianthema triquetra can be exploited as an alternative remedy for cancer, oxidative stress related disorders and in various skin diseases.