OPTIMIZATION OF AMINO ACID PRODUCTION FROM CHICKEN FEATHER BY BACILLUS AMYLOLIQUEFACIENS USING RESPONSE SURFACE METHODOLOGY AS A NEW TOOL FOR POWDERY MILDEW DISEASE BIOCONTROL

The present work aimed to optimize amino acids production by Bacillus amyloliquefaciens 35s from chicken feather using response surface methodology (RSM) and applying the produced amino acids in biological control of powdery mildew disease. Variables affecting amino acids production from B. amyloliquefaciens 35s were screened using Plackett Burman design. Factors showed to be significant to produce amino acids were optimized using central composite design (CCD) of RSM at three coded levels (-1, 0, +1). All obtained data were analyzed by ANOVA with post hoc multiple comparison analysis performed using Tukey’s HSD. TGY (tryptone, glucose, yeast extract) was the basal medium. Impacts of medium components were studied using Plackett-Burman design. “Design Expert 12” StateEase was used to analyze the experimental Plackett- Burman design. Feather, yeast extract, CaCO3, KH2PO4 and agitation rate (using shake flask) were optimized statistically by the CCD design of the RSM. Validation of statistical model of the medium components was done by carrying out the experiment at optimum conditions of the process parameters as determined by the model. Amino acids types were identified by amino acid analyzer. Among the significant medium components, feather, yeast extract, CaCO3, KH2PO4 and agitation rate showed to have significant effect on amino acids production. Predicted maximum amino acids production was observed near the mid-point concentrations of the mentioned above factors and the experimental value (6.55 g/l) was very close to the predicted value of (6.60 g/l) predicted by the model. Amino acid analyzer analysis for the produced amino acids shows the release of asparagine, threonine, serine, glutamine, proline, glycine, alanine, cysteine, valine, methionine, isoleucine, leucine, tyrosine, phenyl alanine, histidine, lysine and arginine, respectively. Effect of chicken feather hydrolysate on cucumber powdery mildew disease was tested by applying foliar application with different concentrations (0, 5, 10, 15, and 20% v/v), then degree of infection and sporulation on cucumber plants were assessed. The lowest concentration of chicken feather hydrolysate showed the lowest effect on disease index. Conversely, cucumber resistance increased by the increasing of feather’s hydrolysate concentrations associated with a clear reduction in the disease index and sporulation. Aside from, extra foliar application showed a decrease in the disease index for all concentrations compared with untreated plants. Foliar application with 20% concentration of chicken feather hydrolysate had the highest stimulation of the studied oxidative enzymes i.e., peroxidase (POX), phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO) and superoxide dismutase (SOD)

ISOLATION AND IDENTIFICATION OF ANTIBIOTIC PRODUCING PSEUDOMONAS FLUORESCENS NBRC-14160 FROM DELTA SOIL IN EGYPT

Pseudomonas fluorescens is one of plant Growth Promoting Rhizobacteria (PGPR) members which has a major role in the biological control of bacterial and fungal pathogens. A research was conducted at the Laboratory of Microbiology, Faculty of Agriculture, Ain Shams University, Qalubyia governorate intended for isolating Pseudomonas fluorescens isolate, efficient in antibiotic production. For isolation, soil samples, collected from Faculty of agriculture farm soil at depth of 10 cm, were screened for Ps. fluorescens isolates by cultivating the samples on King’s medium. Out of 30 isolates obtained, one was selected based on its high lipid content, because of the lipid’s correlation to antibiotic production and inhibitory activity. The selected isolate was characterized by morphological, physiological, biochemical tests then confirmed its identity by 16S rRNA gene sequence analysis and named Pseudomonas fluorescens NBRC-14160 16srRNA. Morphological features showed that Ps. fluorescens NBRC-14160 has large colonies, with irregular surface, opaque, producing green fluorescent pigments. Cells are short rods, Gram negative. Physiological features indicated that Ps. fluorescens NBRC-14160 is capable of producing several exoenzymes including lipase, phospholipase, protease and chitinase and incapable of producing amylase and cellulase. Carbohydrate fermentation tests were positive for fructose, glucose, D-glucose, and galactose, forming acids after 24 hrs. of incubation at 30˚C. However, it loses the ability to ferment inositol, mannose, xylose, mannitol, raffinose and rhamnose sugars. It’s lipid content was 607 mg/g. Inhibitory activity was studied by the method of disc diffusion test against nine pathogenic bacterial and fungal strains. Staphylococcus aureus was the most sensitive bacterial pathogen towards P. fluorescens NBRC-14160 with a 4.5 cm zone of inhibition, while Serratia marscens had 0.95 cm zone. Aspergillus niger and Alternaria solani were the most sensitive fungal pathogens towards P. fluorescens NBRC-14160 with 2.55 and 2.5 cm of inhibition zone, respectively. However, the most resistant fungus was Fusarium oxysporum with 1.3 cm inhibition zone

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

Impact of opioid-free analgesia on pain severity and patient satisfaction after discharge from surgery: multispecialty, prospective cohort study in 25 countries

Background: Balancing opioid stewardship and the need for adequate analgesia following discharge after surgery is challenging. This study aimed to compare the outcomes for patients discharged with opioid versus opioid-free analgesia after common surgical procedures.Methods: This international, multicentre, prospective cohort study collected data from patients undergoing common acute and elective general surgical, urological, gynaecological, and orthopaedic procedures. The primary outcomes were patient-reported time in severe pain measured on a numerical analogue scale from 0 to 100% and patient-reported satisfaction with pain relief during the first week following discharge. Data were collected by in-hospital chart review and patient telephone interview 1 week after discharge.Results: The study recruited 4273 patients from 144 centres in 25 countries; 1311 patients (30.7%) were prescribed opioid analgesia at discharge. Patients reported being in severe pain for 10 (i.q.r. 1-30)% of the first week after discharge and rated satisfaction with analgesia as 90 (i.q.r. 80-100) of 100. After adjustment for confounders, opioid analgesia on discharge was independently associated with increased pain severity (risk ratio 1.52, 95% c.i. 1.31 to 1.76; P < 0.001) and re-presentation to healthcare providers owing to side-effects of medication (OR 2.38, 95% c.i. 1.36 to 4.17; P = 0.004), but not with satisfaction with analgesia (beta coefficient 0.92, 95% c.i. -1.52 to 3.36; P = 0.468) compared with opioid-free analgesia. Although opioid prescribing varied greatly between high-income and low- and middle-income countries, patient-reported outcomes did not.Conclusion: Opioid analgesia prescription on surgical discharge is associated with a higher risk of re-presentation owing to side-effects of medication and increased patient-reported pain, but not with changes in patient-reported satisfaction. Opioid-free discharge analgesia should be adopted routinely

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

Optimization of Polyunsaturated Fatty Acids (PUFAs) Production from Olive Cake by Pseudomonas fluorescens NBRC14160 using Response Surface Methodology and their Application in Kareish Cheese Manufacture

The aim of this study was to optimize the production of polyunsaturated fatty acids (PUFAs), using the response surface methodology (RSM), to use the produced PUFAs in kareish cheese processing. PlackettBurman design (PBD) was employed to screen media components that affect PUFAs development (glucose, olive cake, yeast extract, tryptone, MgSO4, KH2PO4, NH4Cl, agitation speed, incubation time, and pH), and results showed that olive cake and yeast extract, with confidence level > 98%, had a positive effect on PUFAs production. The central composite design (CCD) of the response surface methodology was used to optimize the selected parameters levels where maximum PUFAs production (1790 mg/l) was observed near the mid-point (0) values (concentrations) of olive cake (15 g/l), tryptone (7.5 g/L) and KH2PO4 (1.25 g/l). Polyunsaturated fatty acids account for 47.83 % of the total fatty acid profile, according to gas chromatography analysis of the collected PUFAs. The produced PUFAs was encapsulated using whey protein concentrate and maltodextrin, freeze dried, grinded and incorporated in Kareish cheese manufacture. The average particle size of a 0.005% suspension of oil microcapsules was 671.4 nm with a poly dispersity index of 0.611 indicating a moderate stability of the emulsion. The negative zeta potential of the microcapsules particles was -37.6 mv, which is identical to the -42 mv value recorded in the literature for oil emulsions stabilized by whey protein, maltodextrin, and K-carrageenan. The addition of 0.5 % PUFAs-containing microcapsules to Kareish cheese increased antioxidative activity to 38.13 % compared to 30.14 % for the control, as well as Texture profile analysis (TPA) parameters including hardness, cohesiveness, gumminess, and chewiness. The elasticity of the Kareish cheese sample increased slightly by the addition of 0.5% microcapsules, but higher concentration tended to change the elasticity to a brittleness of the cheese structure

Promoting of Abiotic Stress–Induced Resistance Using Poly-β- Hydroxybutyrate (PHB) By Rhizobium phaseoli In Common Bean Plants

In total, 50 Rhizobium isolates were isolated from the mature root nodules of common beans plants (Phaseolus vulgaris) grown in different nine governorates of Egypt. PHB was optimized by the identified strain using response surface methodology. A total of 11 parameters (pH, incubation period, inoculum size, temperature, agitation speed, mannitol, sucrose, yeast extract, glycine, K2HPO4, and MgSO4) were analyzed for their significant effects on PHB production by the Plackett–Burman design (PBD). Sucrose, yeast extract, glycine, and MgSO4 were the main significant factors affecting PHB accumulation. Central composite design (CCD) of the response surface methodology was used to determine the optimum levels of the selected factors. Rhizobium phaseoli reached the maximum production (4.997 g/L) at run 36 in the presence of 25 g/L of sucrose, 0.0 g/L of yeast extract, 0.87 g/L of glycine, 0.3 g/L of MgSO4, and 5% of inoculation size. In vitro experiments were carried out to test the effect of different stress conditions (pH: 6–11, temperature: 5°C–50°C, salinity: 0.01%–7%, and drought: 0%–5% w/v) on the growth of Rhizobium phaseoli. The results showed that Rhizobium phaseoli can withstand 3% –5% NaCl, high temperature of 30°C– 45°C, alkalinity at pH value of 8 – 10, and drought stress at 3% – 5% w/v polyethylene glycol with growth loss of 50% when grown on modified medium and 75% when grown on the basal one. In vivo experiments were done to study the effect of drought stress levels on the growth parameters of common bean plants. In general, all the treatments with Rhizobium phaseoli grown on the modified medium were superior to Rhizobium phaseoli grown on the basal medium. Also, they showed high tolerance of drought conditions

Enhancing durability and sustainable preservation of Egyptian stone monuments using metabolites produced by Streptomyces exfoliatus

Abstract Despite their threatens for Egyptian stone monuments, A few studies focused on using biocontrol agents against deteriorative fungi and bacteria instead of using chemical assays that leave residuals leading to human toxicity and environmental pollution. This work aims to isolate and identify fungal and bacterial isolates that showed deteriorative activities from stone monuments in Temple of Hathor, Luxor, Egypt, as well as determine the inhibitory activity of metabolites produced by Streptomyces exfoliatus SAMAH 2021 against the identified deteriorative fungal and bacterial strains. Moreover, studying the spectral analysis, toxicological assessment of metabolites produced by S. exfoliatus SAMAH 2021 against health human cell fibroblast, and colorimetric measurements on the selected stone monuments. Ten samples were collected from Temple of Hathor, Luxor, Egypt. Three fungal isolates and one bacterial isolate were obtained and identified as A. niger isolate Hathor 2, C. fioriniae strain Hathor 3, P. chrysogenum strain HATHOR 1, and L. sphaericus strain Hathor 4, respectively. Inhibitory potential of the metabolites in all concentrations used (100–25%) against the recommended antibiotics (Tetracycline 10 µg/ml and Doxycycline (30 µg/ml) showed an inhibitory effect toward all tested deteriorative pathogens with a minimum inhibition concentration (MIC) of 25%. Cytotoxicity test confirmed that microbial filtrate as the antimicrobial agent was safe for healthy human skin fibroblast with IC50 of < 100% and cell viability of 97%. Gas chromatography analysis recorded the existence of thirteen antimicrobial agents, Cis-vaccenic acid; 1,2-Benzenedicarboxylic acid; ç-Butyl-ç-butyrolactone and other compounds. Colorimetric measurements confirmed no color or surface change for the limestone-treated pieces. The use of the metabolite of microbial species antimicrobial as a biocontrol agent raises contemporary issues concerning the bio-protection of the Egyptian monuments to reduce chemical formulas that are toxic to humans and pollute the environment. Such serious problems need further investigation for all kinds of monuments

Utilization of biosynthesized silver nanoparticles from Agaricus bisporus extract for food safety application: synthesis, characterization, antimicrobial efficacy, and toxicological assessment

Abstract The emergence of antimicrobial resistance in foodborne bacterial pathogens has raised significant concerns in the food industry. This study explores the antimicrobial potential of biosynthesized silver nanoparticles (AgNPs) derived from Agaricus bisporus (Mushroom) against foodborne bacterial pathogens. The biosynthesized AgNPs were characterized using various techniques, including UV–visible spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, high-resolution scanning electron microscopy with energy dispersive X-ray spectroscopy, dynamic light scattering, and zeta potential analysis. The antibacterial activity of the AgNPs was tested against a panel of foodborne bacterial strains, and their cytotoxicity was evaluated on normal human skin fibroblasts. Among the tested strains, Pseudomonas aeruginosa ATCC 27853 showed the highest sensitivity with an inhibition zone diameter (IZD) of 48 mm, while Klebsiella quasipneumoniae ATTC 700603 and Bacillus cereus ATCC 11778 displayed the highest resistance with IZDs of 20 mm. The silver cations released by AgNPs demonstrated strong bactericidal effects against both Gram-positive (G + ve) and Gram-negative (G − ve) bacteria, as evidenced by the minimum inhibitory concentration/minimum bactericidal concentration (MBC/MIC) ratio. Moreover, cytotoxicity testing on normal human skin fibroblasts (HSF) indicated that AgNPs derived from the mushroom extract were safe, with a cell viability of 98.2%. Therefore, AgNPs hold promise as an alternative means to inhibit biofilm formation in the food industry sector

Surgical site infection after gastrointestinal surgery in children : an international, multicentre, prospective cohort study

Introduction Surgical site infection (SSI) is one of the most common healthcare-associated infections (HAIs). However, there is a lack of data available about SSI in children worldwide, especially from low-income and middle-income countries. This study aimed to estimate the incidence of SSI in children and associations between SSI and morbidity across human development settings. Methods A multicentre, international, prospective, validated cohort study of children aged under 16 years undergoing clean-contaminated, contaminated or dirty gastrointestinal surgery. Any hospital in the world providing paediatric surgery was eligible to contribute data between January and July 2016. The primary outcome was the incidence of SSI by 30 days. Relationships between explanatory variables and SSI were examined using multilevel logistic regression. Countries were stratified into high development, middle development and low development groups using the United Nations Human Development Index (HDI). Results Of 1159 children across 181 hospitals in 51 countries, 523 (45 center dot 1%) children were from high HDI, 397 (34 center dot 2%) from middle HDI and 239 (20 center dot 6%) from low HDI countries. The 30-day SSI rate was 6.3% (33/523) in high HDI, 12 center dot 8% (51/397) in middle HDI and 24 center dot 7% (59/239) in low HDI countries. SSI was associated with higher incidence of 30-day mortality, intervention, organ-space infection and other HAIs, with the highest rates seen in low HDI countries. Median length of stay in patients who had an SSI was longer (7.0 days), compared with 3.0 days in patients who did not have an SSI. Use of laparoscopy was associated with significantly lower SSI rates, even after accounting for HDI. Conclusion The odds of SSI in children is nearly four times greater in low HDI compared with high HDI countries. Policies to reduce SSI should be prioritised as part of the wider global agenda.Peer reviewe