Eco-friendly remediation of tetracycline antibiotic from polluted water using waste-derived surface re-engineered silica sand.

A new green reactive adsorbent (calcium ferric oxide silica sand (CFO-SS)) made from wastepaper sludge ash and ferric ions was synthesised and shown to remove tetracycline antibiotics (TC) from contaminated water effectively. The synthesised sand was dried at 95 °C, and a series of batch and fixed bed experiments were performed to determine the optimum operating conditions. Results showed that the adsorption capacity of the CFO-SS increases with the concentration gradient between the solid and liquid phases. 0.3 g of the new adsorbent was proven sufficient to remove more than 90% of the TC at a pollutant dose of 50 mg/L in 50 mL of simulated groundwater with an agitation speed of 200 rpm for 3 h. The adsorption isotherm followed the Langmuir isotherm model, with a loading capacity of 21.96 mg/g at pH 7, while the Pseudo second-order model best described the absorption kinetics. The adsorption mechanisms proposed included electrostatic interaction, intraparticle diffusion, hydrogen bonding, and cation-π interactions. Characterisation investigations revealed that the newly precipitated oxides on silica sand play an essential role in TC adsorption support. In fixed-bed experiments, it was discovered that reducing the flow rate and inflow concentration of TC and increasing the sorbent mass significantly extended the lifetime of the produced sorbent in the packed column. The measured breakthrough curves were best fit with the Adams-Bohart and the Clark models, as they provided the highest square root number (R2) values. Finally, considering the efficacy of CFO-SS in TC adsorption performance, it can be noted that the novel synthesised reactive material is an efficient and environmentally friendly material for TC removal, and it presents a potential solution to resolving the challenge of TC-rich groundwater.

Cladophialophora bantiana metabolites are efficient in the larvicidal and ovicidal control of Aedes aegypti, and Culex quinquefasciatus and have low toxicity in zebrafish embryo.

Mosquitoes' current insecticide resistance status in available public health insecticides is a serious threat to mosquito control initiatives. Microbe-based control agents provide an alternative to conventional pesticides and insecticides, as they can be more targeted than synthetic insecticides. The present study was focused on identifying and investigating the mosquitocidal potential of Cladophialophora bantiana, an endophytic fungus isolated from Opuntia ficus-indica. The Cladophialophora species was identified through phylogenetic analysis of the rDNA sequence. The isolated fungus was first evaluated for its potential to produce metabolites against Aedes aegpti and Culex quinquefasciatus larvae in the 1-4th instar. The secondary metabolites of mycelium extract were assessed at various test doses (100, 200, 300, 400, and 500 µg/mL) in independent bioassays for each instar of selected mosquito larvae. After 48 h of exposure, A. aegypti expressed LC50 values of 13.069, 18.085, 9.554, and 11.717 µg/mL and LC90 = 25.702, 30.860, 17.275, and 19.601 µg/mL; followed by C. quinquefasciatus LC50 = 14.467, 11.766, 5.934, and 7.589 µg/mL, and LC90 = 29.529, 20.767, 11.192, and 13.296 µg/mL. The mean % of ovicidal bioassay was recorded 120 h after exposure. The hatchability (%) was proportional to mycelia metabolite concentration. The enzymatic level of acetylcholinesterase in fungal mycelial metabolite treated 4th instar larvae indicated a dose-dependent pattern. The GC-MS profile of C. bantiana extracts identified five of the most abundant compounds, namely cyclobutane, trans-3-undecene-1,5-diyne, 1-bromo-2-chloro, propane, 1,2,3-trichloro-2-methyl-, 5,5,10,10-tetrachlorotricyclo, and phenol, which had the killing effect in mosquitoes. Furthermore, the C. bantiana fungus ethyl acetate extracts had a strong larvicidal action on A. aegypti and C. quinquefasciatus. Finally, the toxicity test on zebrafish embryos revealed the induction of malformations only at concentrations above 1 mg/mL. Therefore, our study pioneered evidence that C. bantiana fungal metabolites effectively control A. aegypti and C. qunquefasciastus and show less lethality in zebrafish embryos at concentrations up to 500 µg/mL.

Microtiter plate with built-in oxygen sensors: a novel approach to investigate the dynamics of Pseudomonas aeruginosa growth suppression in the presence of divalent cations and antibiotics.

The depletion of dissolved oxygen in a defined synthetic medium can be measured in real time, using a micro-well plate format, associated with a fluorescent plate reader. This technology is appropriate for investigating the effect of antibiotics on cell kinetics because there is a direct correlation between the latter and the amount of dissolved oxygen in the medium of an assay. In this study, the metabolic activity of the opportunistic human pathogen Pseudomonas aeruginosa PA01 was investigated using the OxoPlate OP96U optical sensor technology. The response of P. aeruginosa to aminoglycoside antibiotics when Ca2+and Mg2+ ions are present in the Evans defined synthetic medium was measured. The results revealed that the effect of antibiotics on P. aeruginosa is influenced by the concentration of divalent cations present in the test medium, although the efficiency of Ca2+ in supressing antibiotic activity was found to be greater than that of Mg2+. By comparison to tobramycin, the effect of amikacin is largely inhibited by the Ca2+and Mg2+concentrations. The study results underscore that the reliability of the observation of growth inhibitors is enhanced by the oxygen consumption measurements. Thus, the OxoPlate OP96U system is proven to be an accurate method to test the effectiveness of antibiotic treatments against P. aeruginosa.

Biomineralisation to Increase Earth Infrastructure Resilience.

The vulnerability of buildings and structures to rain and flooding due to a lack of adaptive capacity is an issue all over the world. Exploring the bio-resources availability and engineering performance is crucial to increase infrastructure's resilience. The current study analyses earth-based mortars using mineral precipitation as a biostabiliser (bio) and compares their performance with cement-based mortars. Cultures of S. oneidensis with a concentration of 2.3 × 108 cfu/mL were used to prepare earth-based and cement-based mortars with a ratio of 6% of binder. Microstructure analyses through SEM/EDS, water absorption, moisture buffering, mechanical strength, and porosity are discussed. The biostabiliser decreases water absorption in tidal-splash and saturated environments for earth and cement mortars due to calcium carbonate precipitation. The biostabiliser can prevent water migration more effectively for the cement-based (60% reduction) than for the earth-based mortars (up to 10% reduction) in the first 1 h of contact with water. In an adsorption/desorption environment, the conditions favour desorption in cem+bio, and it seems that the biostabiliser precipitation facilitates the release of the chemicals into the mobile phase. The precipitation in the earth+bio mortar porous media conditions favours the adsorption of water molecules, making the molecule adhere to the stationary phase and be separated from the other sample chemicals. The SEM/EDS performed for the mortars confirms the calcium carbonate precipitation and shows that there is a decrease in the quantity of Si and K if the biostabiliser is used in cement and earth-mortars. This decrease, associated with the ability of S. oneidensis to leach silica, is more impressive for earth+bio, which might be associated with a dissolution of silicate structures due to the presence of more water. For the tested earth-based mortars, there was an increase of 10% for compressive and flexural strength if the biostabiliser was added. For the cement-based mortars, the strength increase was almost double that of the plain one due to the clay surface negative charge in the earth-based compositions.

Persister cells: formation, resuscitation and combative therapies.

Persister cells, or superfits, have been strongly implicated in the recalcitrance and recurrence of chronic bacterial infection through the dormant (metabolically reduced) phenotype they display and the tolerance to antimicrobial agents this dormancy grants them. The complex biochemical events that lead to the formation of persister cells are not completely understood, though much research has linked the degradation of type II toxin/antitoxin systems and reduced cellular ATP levels to the rise in stress response molecules (where (p)ppGpp is of particular interest), which induce this dormant state. The equally complex mechanism of resuscitation is initiated by the cells' ability to sense nutrient availability via chemotaxis systems. Levels of secondary messenger proteins (i.e., cAMP) within the cell are reduced to allow the resuscitation of ribosomes, by ribosomal resuscitation factor HflX, to reinstate protein synthesis and, therefore, growth to re-populate. Techniques of superfit eradication utilise one, or more, of three approaches (i) direct killing, (ii) re-sensitising persister cells to conventional antimicrobials, or (iii) prevention of persister formation though few laboratory findings have been translated to clinical practice. This work will outline current findings in the field with a critical approach, where possible.

A Comprehensive Review for Groundwater Contamination and Remediation: Occurrence, Migration and Adsorption Modelling.

The provision of safe water for people is a human right; historically, a major number of people depend on groundwater as a source of water for their needs, such as agricultural, industrial or human activities. Water resources have recently been affected by organic and/or inorganic contaminants as a result of population growth and increased anthropogenic activity, soil leaching and pollution. Water resource remediation has become a serious environmental concern, since it has a direct impact on many aspects of people's lives. For decades, the pump-and-treat method has been considered the predominant treatment process for the remediation of contaminated groundwater with organic and inorganic contaminants. On the other side, this technique missed sustainability and the new concept of using renewable energy. Permeable reactive barriers (PRBs) have been implemented as an alternative to conventional pump-and-treat systems for remediating polluted groundwater because of their effectiveness and ease of implementation. In this paper, a review of the importance of groundwater, contamination and biological, physical as well as chemical remediation techniques have been discussed. In this review, the principles of the permeable reactive barrier's use as a remediation technique have been introduced along with commonly used reactive materials and the recent applications of the permeable reactive barrier in the remediation of different contaminants, such as heavy metals, chlorinated solvents and pesticides. This paper also discusses the characteristics of reactive media and contaminants' uptake mechanisms. Finally, remediation isotherms, the breakthrough curves and kinetic sorption models are also being presented. It has been found that groundwater could be contaminated by different pollutants and must be remediated to fit human, agricultural and industrial needs. The PRB technique is an efficient treatment process that is an inexpensive alternative for the pump-and-treat procedure and represents a promising technique to treat groundwater pollution.

Design Service Life of RC Structures with Self-Healing Behaviour to Increase Infrastructure Carbon Savings.

Corrosion of reinforced concrete (RC) structures costs the UK GBP 23b annually and is one of the main durability problems contributing to the development of rust, spalling, cracking, delamination, and structural deterioration. This paper intends to demonstrate the benefit of using tailored self-healing bacteria-based concrete for RC corrosion minimisation and service life increase. The purpose was to evaluate the enhancement in the lifespan of the structure exposed to a harsh marine microenvironment by utilising a probabilistic performance-based method. Comparison is made with the performance of a commercially available solution and in terms of embodied carbon impact. Three different concretes, using CEM I 52.5N, CEM II/A-D, and CEM III/A, were tested with and without an iron-respiring bioproduct (BIO) and an added admixture corrosion inhibitor (AACI). Results show that bioproduct significantly contributes to service life increase of RC structures with CEMIII/A. The repair solution with self-healing behaviour not only increases RC service life, but also enables us to decrease the required cover thickness from 60 mm to 50 mm in an XS2 chloride environment. In both XS2 and XS3 environments, a comparison of CEMIII/A+BIO and CEMII/A-D+AACI concrete shows the benefit of using bioproduct in corrosion inhibition context, besides contributing to an embodied carbon reduction of more than 20%.

Nitro-Carba test, a novel and simple chromogenic phenotypic method for rapid screening of carbapenemase-producing Enterobacteriaceae.

OBJECTIVES: In this study, a rapid and simple chromogenic method for screening of carbapenemase-producing Enterobacteriaceae (CPE), namely the Nitro-Carba test (NCT), was developed. METHODS: The NCT was validated using a total of 31 carbapenemase-producing isolates [9 Klebsiella pneumoniae carbapenemase (KPC), 11 metallo-ß-lactamase (MBL) and 11 OXA-48] and 56 non-carbapenemase-producing isolates. The assay relies on the hydrolysis of nitrocefin by carbapenemases in the presence of carbapenem antibiotics. Carbapenemases were extracted with lysis buffer prior to addition to wells with and without imipenem (IPM), meropenem (MEM) and ertapenem (ETP). Following addition of nitrocefin, a change in colour from yellow to red, indicating carbapenemase production, was observed within 20min. The susceptibility profiles of each bacterial isolate were also investigated. RESULTS: The NCT detected all 31 CPE within a timeframe of only 10s to 12min. All carbapenemase-producers hydrolysed nitrocefin in all wells. No colour change in wells with carbapenems was observed in non-carbapenemase-producers. The sensitivity for all three carbapenems was 100%, whilst the specificity of IPM, MEM and ETP was 64.3%, 91.1% and 100%, respectively. IPM, MEM and ETP had minimum inhibitory concentrations (MICs) against all carbapenemase-producing strains ranging from 0.5µg/mL to ≥256µg/mL, 0.25µg/mL to ≥256µg/mL and 1µg/mL to ≥256µg/mL, respectively. OXA-48-producing isolates showed lower MICs compared with MBL- and KPC-producing isolates. CONCLUSION: This assay is a promising method for detecting CPE rapidly. The NCT is a simple and reliable method capable of detecting CPE even in carbapenem-susceptible strains.

A nitrocefin disc supplemented with ertapenem for rapid screening of carbapenemase-producing Enterobacteriaceae.

Reliable, simple and rapid methods for laboratory detection of carbapenemases are important for an appropriate antibiotic administration. A nitrocefin disc containing ertapenem for rapid screening of carbapenemase production among Enterobacteriaceae is developed in the present study. A total of 87 molecularly-confirmed Enterobacteriaceae including 31 carbapenemase producers and 56 non-carbapenemase producers, were tested with nitrocefin discs supplemented with and without ertapenem (20 µg/disc). Nitrocefin discs with ertapenem successfully discriminated all 31 carbapenemase and all non-carbapenemase producers within 30 minutes. The sensitivity and specificity of the method were 100%. The minimum inhibitory concentrations (MICs) of ertapenem against all carbapenemase-producing isolates ranged from 1 to ≥256 µg/mL. This simple test could help to minimize the treatment failure and control the dissemination of infections caused by carbapenemase-associated resistant bacteria. It is a promising approach that could be performed routinely in any laboratory.

The performance of a resazurin chromogenic agar plate with a combined disc method for rapid screening of extended-spectrum-ß-lactamases, AmpC ß-lactamases and co-ß-lactamases in Enterobacteriaceae.

A promising means of rapid screening of extended-spectrum-ß-lactamase (ESBL), AmpC ß-lactamase, and co-production of ESBL and AmpC that combines resazurin chromogenic agar (RCA) with a combined disc method is here reported. Cefpodoxime (CPD) discs with and without clavulanic acid (CA), cloxacillin (CX) and CA+CX were evaluated against 86 molecularly confirmed ß-lactamase-producing Enterobacteriaceae, including 15 ESBLs, 32 AmpCs, nine co-producers of ESBL and AmpC and 30 carbapenemase producers. The CA and CX synergy test successfully detected all ESBL producers (100% sensitivity and 98.6% specificity) and all AmpC producers (100% sensitivity and 96.36% specificity). This assay also performed well in screening for co-existence of ESBL and AmpC (88.89% sensitivity and 100% specificity). The RCA assay is simple and inexpensive and provides results within 7 hr. It can be performed in any microbiological laboratory, in particular, in geographic regions in which ESBL, AmpC or co-ß-lactamase-producing Enterobacteriaceae are endemic.

A demonstration of athermal effects of continuous microwave irradiation on the growth and antibiotic sensitivity of Pseudomonas aeruginosa PAO1.

Stress, caused by exposure to microwaves (2.45 GHz) at constant temperature (37 ± 0.5°C), alters the growth profile of Pseudomonas aeruginosa PAO1. In the absence of microwave treatment a simple, highly reproducible growth curve was observed over 24 h or more. Microwave treatment caused no reduction in growth during the first 6 h, but at a later stage (>12 h) the growth was markedly different to the controls. Secondary growth, typical of the presence of persisters clearly became apparent, as judged by both the dissolved oxygen and the cell density profiles. These treated cells showed distinct morphological changes, but on regrowth these cells reverted to normal. The microwave induced persisters were subject to antibiotic challenge (tobramycin) and showed increased sensitivity when compared to the unstressed planktonic cells. This is in marked contrast to antibiotic induced persisters which show increased resistance. This provides evidence for both a nonthermal effect of microwaves and a previously undescribed route to a novel form of antibiotic susceptible persister cells. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:37-44, 2017.

Phenotypic detection of AmpC ß-lactamases, extended-spectrum ß-lactamases and metallo-ß-lactamases in Enterobacteriaceae using a resazurin microtitre assay with inhibitor-based methods.

Dissemination of antibiotic resistance in Enterobacteriaceae mediated by AmpC ß-lactamase, extended-spectrum ß-lactamase (ESBL) and metallo-ß-lactamase (MBL) is clinically significant. A simple and relatively quick method for the detection of these resistance phenotypes would greatly improve chemotherapeutic recommendation. This technology would provide valuable input in our surveillance of resistance on a global stage, particularly if the methodology could be applicable to resource-poor settings. A resazurin microtitre plate (RMP) assay incorporating cloxacillin, clavulanic acid and EDTA for the rapid phenotypic identification of AmpC, ESBL and MBL and the co-existence of ß-lactamases has been developed. A total of 47 molecularly characterized Enterobacteriaceae clinical isolates producing AmpCs, ESBLs, co-producers of ESBL and AmpC, MBLs and co-producers of ESBL and MBL were phenotypically examined using the RMP assay. The ceftazidime- and cefotaxime-based RMP assays successfully detected all 16 AmpC, 14 ESBL and 9 MBL producers, 6 ESBL-AmpC co-producers and 2 ESBL-MBL co-producers without false-positive results. The ceftazidime-based assay was more reliable in detecting AmpC alone, while the cefotaxime-based assay performed better in identifying co-producers of ESBL and AmpC. There was no difference in the detection of ESBL and MBL producers. The findings of the present study suggest that use of the RMP assay with particular ß-lactamase inhibitors explicitly detects three different ß-lactamases, as well as co-existence of ß-lactamases, within 6 h of initial isolation of the pathogen. This assay is applicable to carry out in any laboratory, is cost-effective and is easy to interpret. It could be implemented in screening patients and controlling infection and for surveillance purposes.

Incorporation of L-lysine and 2,2'-dipyridyl in the growth media promotes desferrioxamine E production by an actinobacterium.

The study describes the use of the chelating agent 2,2'-dipyridyl in conjunction with lysine to increase the production of the siderophore desferrioxamine E by a previously described actinobacterium 23F. Desferrioxamine E is a type of siderophore known to be produced by Streptomycete species. Lysine is a precursor of the siderophore and its presence in the culture medium is known to promote desferrioxamine E synthesis. The further addition of 2,2'-dipyridyl was found to enhance production of the siderophore in the presence of lysine (5 g l(-1)) nearly twofold when incorporated at a concentration of 200 µM. Increasing the concentration of the chelating agent above 200 µM resulted in a decrease in siderophore production. The role of the chelating agent was thought to be in creating iron-limiting conditions in the culture medium and so promoting the induction of the desferrioxamine E biosynthetic pathway. This medium is likely to be a useful tool in the screening for producers of desferrioxamine E.

Real-Time Monitoring of Pseudomonas Aeruginosa Concentration Using a Novel Electromagnetic Sensors Microfluidic Cell Structure.

This study demonstrates an electromagnetic wave-based sensor embedded within a fluidic cell for the purposes of quantifying Pseudomonas aeruginosa in real time, which implies it could be applied for provision of point-of-care diagnostics. The sensors operates through the interaction of the electromagnetic field with the analyte flowing through the fluidic system, and via the sensor head which has a specifically designed planar pattern to maximize the sensor sensitivity for the given bacteria type. The sensor is demonstrated to respond linearly (R(2) = 0.9942) to OD(550) 25 × 10(-3) - 1.0 bacteria concentration through changing resonant frequency and peak quality factor. This innovative approach is expected to contribute to better provision of healthcare services, minimizing the need for hospital visits through real-time point-of-care diagnostics as opposed to lengthy laboratory assays.

A new approach to studying ion uptake by actinomycetes.

A streptomycete that had the ability to avidly sequester iron via siderophores was previously isolated from environmental soil samples. The chelating agent expressed by this organism is confirmed by HPLC as desferrioxamine E. Although the traditional chromo azuerol sulphate (CAS) assay for detection of siderophores is based upon the chelation of iron we were interested to examine the relationship of these iron-capturing molecules with other ions. Consequently, a new approach was employed that enabled the assessment of the affinity of the siderophore moieties for other ions by adapting the CAS assay. The present study reveals that the isolate produced a siderophore that was capable of sequestering a range of ions including Mn, Co, Cd, Ni, Al, Li, Cu, Zn and Mg. On the basis of the assay described it would appear that the organism sequesters copper more readily than iron. This raises an age-old debate surrounding the replacement of copper as a fundamentally essential element with iron as a consequence of the evolution of the di-oxygen environment.