Response Surface Methodology for the production of endopolygalacturonase by a novel Bacillus licheniformis

Background: Polygalacturonase is one of the most important commercial pectinase. The production cost and the mesophilic nature of the present polygalacturonase is a big problem in its application in the juice industry. A lot of work is going on for the isolation of thermophilic bacterial strains which can utilize pectin as the only carbon source.Methods: Bacterial strains were isolated from rotten fruits and vegetables and cultured at 50 – 70oC. The strains were than screened for endopolygalacturonase activity and identified on the basis of 16S rRNA sequence. Different growth parameters for the production of endopolygalacturonase by Bacillus licheniformis IEB-8 were optimized using Response Surface Methodology under Center Composite Design using JMP-12 software. Endopolygalacturonase was purified in two steps; ammonium sulfate precipitation and then by size exclusion column chromatography.Results: Only four strains, IEB-8, IEB-11, IEB-12 and IEB-13 showed growth above 60oC. Among these four, only IEB-8 was found to be endopolygalacturonase positive, which was identified as Bacillus licheniformis by 16S rRNA gene sequence. Purification fold of 2.57 and 7.48 in the specific activity were achieved using ammonium sulfate precipitation and gel filtration chromatography respectively. Molecular weight of the purified endopolygalacturonase was found to be 42 kDa. The purified endopolygalacturonase showed an optimum pH of 7 and optimum temperature of 55oC.Conclusion: Bacillus licheniformis IEB-8 is a novel bacteria which can efficiently be utilized in the industry for the production of endopolygalacturonase very cheaply. Furthermore, the high optimum working temperature of endopolygalacturonase, increases its significance for its industrial applications.Keywords: Endopolygalacturonase; Bacillus licheniformis; Thermophilic; Response Surface Methodology; Ammonium sulfate precipitatio

Graphene oxide incorporated polyether sulfone nanocomposite antifouling ultrafiltration membranes with enhanced hydrophilicity

In this study, the polyether sulfone (PES) based membranes containing various concentrations of graphene oxide (GO), polyvinylpyrrolidone (PVP), and polyethylene glycol (PEG) were synthesized via the phase immersion method. This study aims to evaluate the effect of GO addition on the structural properties and performance of the membranes. The membranes were analyzed by x-ray diffraction (XRD), scanning electron microscopy (SEM), and Fourier transforms infrared spectroscopy (FTIR). The FTIR-ATR spectra indicated the presence of hydroxyl and carboxylic acid groups on the surface of GO-incorporated membranes, which improved their dispersion in the polymeric matrix and hydrophilicity. The SEM analysis of the GO-containing PES membranes confirmed the formation of a well-defined finger-like porous structure presenting adequate water flux (95 l.m(-2).h(-1)) and salt rejection (72%) compared to the pristine PES membranes (46 l.m(-2).h(-1) and similar to 35%, respectively). In addition, the significantly large wettability and considerably improved antibacterial characteristic (against S. aureus and E. coli strains) of the GO-PES membranes are considered impressive features.National University of Sciences and Technology (NUST) Research Directorate; HEC; NRPU [6020]6020; Higher Education Commission, Pakistan, HEC; National University of Sciences and Technology, NUS

Antimicrobial resistance among migrants in Europe: a systematic review and meta-analysis

BACKGROUND: Rates of antimicrobial resistance (AMR) are rising globally and there is concern that increased migration is contributing to the burden of antibiotic resistance in Europe. However, the effect of migration on the burden of AMR in Europe has not yet been comprehensively examined. Therefore, we did a systematic review and meta-analysis to identify and synthesise data for AMR carriage or infection in migrants to Europe to examine differences in patterns of AMR across migrant groups and in different settings. METHODS: For this systematic review and meta-analysis, we searched MEDLINE, Embase, PubMed, and Scopus with no language restrictions from Jan 1, 2000, to Jan 18, 2017, for primary data from observational studies reporting antibacterial resistance in common bacterial pathogens among migrants to 21 European Union-15 and European Economic Area countries. To be eligible for inclusion, studies had to report data on carriage or infection with laboratory-confirmed antibiotic-resistant organisms in migrant populations. We extracted data from eligible studies and assessed quality using piloted, standardised forms. We did not examine drug resistance in tuberculosis and excluded articles solely reporting on this parameter. We also excluded articles in which migrant status was determined by ethnicity, country of birth of participants' parents, or was not defined, and articles in which data were not disaggregated by migrant status. Outcomes were carriage of or infection with antibiotic-resistant organisms. We used random-effects models to calculate the pooled prevalence of each outcome. The study protocol is registered with PROSPERO, number CRD42016043681. FINDINGS: We identified 2274 articles, of which 23 observational studies reporting on antibiotic resistance in 2319 migrants were included. The pooled prevalence of any AMR carriage or AMR infection in migrants was 25·4% (95% CI 19·1-31·8; I2 =98%), including meticillin-resistant Staphylococcus aureus (7·8%, 4·8-10·7; I2 =92%) and antibiotic-resistant Gram-negative bacteria (27·2%, 17·6-36·8; I2 =94%). The pooled prevalence of any AMR carriage or infection was higher in refugees and asylum seekers (33·0%, 18·3-47·6; I2 =98%) than in other migrant groups (6·6%, 1·8-11·3; I2 =92%). The pooled prevalence of antibiotic-resistant organisms was slightly higher in high-migrant community settings (33·1%, 11·1-55·1; I2 =96%) than in migrants in hospitals (24·3%, 16·1-32·6; I2 =98%). We did not find evidence of high rates of transmission of AMR from migrant to host populations. INTERPRETATION: Migrants are exposed to conditions favouring the emergence of drug resistance during transit and in host countries in Europe. Increased antibiotic resistance among refugees and asylum seekers and in high-migrant community settings (such as refugee camps and detention facilities) highlights the need for improved living conditions, access to health care, and initiatives to facilitate detection of and appropriate high-quality treatment for antibiotic-resistant infections during transit and in host countries. Protocols for the prevention and control of infection and for antibiotic surveillance need to be integrated in all aspects of health care, which should be accessible for all migrant groups, and should target determinants of AMR before, during, and after migration. FUNDING: UK National Institute for Health Research Imperial Biomedical Research Centre, Imperial College Healthcare Charity, the Wellcome Trust, and UK National Institute for Health Research Health Protection Research Unit in Healthcare-associated Infections and Antimictobial Resistance at Imperial College London

Surgical site infection after gastrointestinal surgery in high-income, middle-income, and low-income countries: a prospective, international, multicentre cohort study

Background: Surgical site infection (SSI) is one of the most common infections associated with health care, but its importance as a global health priority is not fully understood. We quantified the burden of SSI after gastrointestinal surgery in countries in all parts of the world. Methods: This international, prospective, multicentre cohort study included consecutive patients undergoing elective or emergency gastrointestinal resection within 2-week time periods at any health-care facility in any country. Countries with participating centres were stratified into high-income, middle-income, and low-income groups according to the UN's Human Development Index (HDI). Data variables from the GlobalSurg 1 study and other studies that have been found to affect the likelihood of SSI were entered into risk adjustment models. The primary outcome measure was the 30-day SSI incidence (defined by US Centers for Disease Control and Prevention criteria for superficial and deep incisional SSI). Relationships with explanatory variables were examined using Bayesian multilevel logistic regression models. This trial is registered with ClinicalTrials.gov, number NCT02662231. Findings: Between Jan 4, 2016, and July 31, 2016, 13 265 records were submitted for analysis. 12 539 patients from 343 hospitals in 66 countries were included. 7339 (58·5%) patient were from high-HDI countries (193 hospitals in 30 countries), 3918 (31·2%) patients were from middle-HDI countries (82 hospitals in 18 countries), and 1282 (10·2%) patients were from low-HDI countries (68 hospitals in 18 countries). In total, 1538 (12·3%) patients had SSI within 30 days of surgery. The incidence of SSI varied between countries with high (691 [9·4%] of 7339 patients), middle (549 [14·0%] of 3918 patients), and low (298 [23·2%] of 1282) HDI (p < 0·001). The highest SSI incidence in each HDI group was after dirty surgery (102 [17·8%] of 574 patients in high-HDI countries; 74 [31·4%] of 236 patients in middle-HDI countries; 72 [39·8%] of 181 patients in low-HDI countries). Following risk factor adjustment, patients in low-HDI countries were at greatest risk of SSI (adjusted odds ratio 1·60, 95% credible interval 1·05–2·37; p=0·030). 132 (21·6%) of 610 patients with an SSI and a microbiology culture result had an infection that was resistant to the prophylactic antibiotic used. Resistant infections were detected in 49 (16·6%) of 295 patients in high-HDI countries, in 37 (19·8%) of 187 patients in middle-HDI countries, and in 46 (35·9%) of 128 patients in low-HDI countries (p < 0·001). Interpretation: Countries with a low HDI carry a disproportionately greater burden of SSI than countries with a middle or high HDI and might have higher rates of antibiotic resistance. In view of WHO recommendations on SSI prevention that highlight the absence of high-quality interventional research, urgent, pragmatic, randomised trials based in LMICs are needed to assess measures aiming to reduce this preventable complication

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

KINETIC CHARACTERIZATION OF PURIFIED LACCASE PRODUCED FROM Trametes versicolor IBL-04 IN SOLID STATE BIO-PROCESSING OF CORNCOBS

A locally isolated white rot fungal strain Trametes versicolor IBL-04 produced high laccase activities in solid state bio-processing of corn cobs. Addition of glucose and yeast extract (C: N ratio; 25:1) enhanced laccase synthesis. Addition of Tween-80 and CuSO4 enhanced laccase production to 1012 U/mL under optimized process conditions. Laccase was further purified to 2.89-fold (specific activity of 840 U/mg) by ammonium sulfate fractional precipitation, dialysis, and Sephadex G-100 gel filtration chromatography. The purified laccase had a relative molecular mass of 63 kDa as detected by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). Best enzyme activity was at pH 5 and 40oC. Using 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as substrate, the enzyme showed maximum activity (Vmax) of 780 U/mL with a corresponding Michaelis constant (Km) value of 73µM. Among the different activators/inhibitors, Cu2+, Mn2+, and Fe2+ stimulated laccase activity, whereas EDTA and cystein inhibited the enzyme. The higher Vmax and lower Km for T. versicolor IBL-04 laccase as compared to most of the reported laccases suggests its potential for industrial applications