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A Sustainable Future
To highlight the vital role microbiology plays in delivering on the UN Sustainable Development Goals (SDGs), we have created a collection of must-read research on three critical aspects of the SDGs: antimicrobial resistance, soil health, and the circular economy.
Collection Contents
1 - 20 of 86 results
Shotgun proteomic analysis of nanoparticle-synthesizing Desulfovibrio alaskensis in response to platinum and palladiumMore Less
Platinum and palladium are much sought-after metals of critical global importance in terms of abundance and availability. At the nano-scale these metals are of even higher value due to their catalytic abilities for industrial applications. Desulfovibrio alaskensis is able to capture ionic forms of both of these metals, reduce them and synthesize elemental nanoparticles. Despite this ability, very little is known about the biological pathways involved in the formation of these nanoparticles. Proteomic analysis of D. alaskensis in response to platinum and palladium has highlighted those proteins involved in both the reductive pathways and the wider stress-response system. A core set of 13 proteins was found in both treatments and consisted of proteins involved in metal transport and reduction. There were also seven proteins that were specific to either platinum or palladium. Overexpression of one of these platinum-specific genes, a NiFe hydrogenase small subunit (Dde_2137), resulted in the formation of larger nanoparticles. This study improves our understanding of the pathways involved in the metal resistance mechanism of Desulfovibrio and is informative regarding how we can tailor the bacterium for nanoparticle production, enhancing its application as a bioremediation tool and as a way to capture contaminant metals from the environment.
Outer membrane protein I is associated with poly-β-hydroxybutyrate granules and is necessary for optimal polymer accumulation in Azotobacter vinelandii on solid medium
Azotobacter vinelandii is a soil bacterium that is able to synthesize poly-β-hydroxybutyrate (PHB), a polymer used to produce biodegradable plastic. PHB is stored in the cytoplasm as granules surrounded by several proteins such as the major phasin PhbP, PHB synthase and PHB depolymerase, among others. Many studies have reported the presence of membrane proteins on PHB granules due to contamination during the polymer extraction procedures. Previously, the outer membrane protein I (OprI) was detected on the polymer granules in A. vinelandii . In this study, by using random transposon mutagenesis, we identified that a mutation in the oprI gene diminished PHB accumulation in A. vinelandii on solid medium. Electron microscopy confirmed the low polymer production by the oprI mutant. Analysis of PHB granules by Tricine-SDS-PAGE revealed that the absence of OprI affected the protein profile of the granules, suggesting that OprI could have a structural role in A. vinelandii . Thus, some membrane proteins on PHB granules may not be artefacts as previously described.
Restoring the activity of the antibiotic aztreonam using the polyphenol epigallocatechin gallate (EGCG) against multidrug-resistant clinical isolates of Pseudomonas aeruginosa
Introduction. Pseudomonas aeruginosa is an important Gram-negative pathogen that is intrinsically multidrug-resistant (MDR) and frequently associated with healthcare-associated outbreaks. With increasing resistance to antibiotics and with very few novel drugs under development, clinicians often use combinations to treat critically ill patients.
Aim. The aim of this study was to evaluate the ability of epigallocatechin (EGCG) to restore the activity of aztreonam against clinical MDR strains of P. aeruginosa .
Methodology. Checkerboard and time–kill kinetic assays were performed to assess synergy in vitro and the Galleria mellonella model of infection was used to test the efficacy of the combination in vivo. Accumulation assays were performed to gain insight into the mechanism of action.
Results. The results demonstrate that synergy between aztreonam and EGCG exists [fractional inhibitory concentration indices (FICIs) 0.02-0.5], with the combination affording significantly (P=<0.05) enhanced bacterial killing, with a >3 log10 reduction in colony-forming units ml−1 at 24 h. EGCG was able to restore susceptibility to aztreonam to a level equal to or below the breakpoint set by the European Committee for Antimicrobial Susceptibility Testing. In G. mellonella, the combination was superior to monotherapy, with increased larval survival observed (94 % vs ≤63 %). We also demonstrated the relatively low toxicity of EGCG to human keratinocytes and G. mellonella larvae. Accumulation assay data suggest that the mechanism of synergy may be due to EGCG increasing the uptake of aztreonam.
Conclusion. EGCG was able to restore the activity of aztreonam against MDR P. aeruginosa . The data presented support further evaluation of the aztreonam–EGCG combination and highlight its potential for use in clinical medicine.
Isolation and characterization of novel soil- and plant-associated bacteria with multiple phytohormone-degrading activities using a targeted methodologyMore Less
Ethylene (ET), salicylic acid (SA) and indole-3-acetic acid (IAA) are important phytohormones regulating plant growth and development, as well as plant-microbe interactions. Plant growth-promoting bacteria (PGPB) naturally associate with plants and facilitate plant growth through a variety of mechanisms, including the ability to modulate the concentrations of these phytohormones in planta. Importantly, the wide presence of phytohormone degradation mechanisms amongst symbiotic and other soil- and plant-associated bacteria indicates that the ability to modulate phytohormone concentrations plays an important role in bacterial colonization and plant-growth promotion abilities. Obtaining phytohormone-degrading bacteria is therefore key for the development of novel solutions aiming to increase plant growth and protection. In this paper, we report an optimized targeted methodology and the consequent isolation of novel soil- and plant-associated bacteria, including rhizospheric, endophytic and phyllospheric strains, with the ability to degrade the phytohormones, SA and IAA, as well as the ET precursor, 1-aminocyclopropane-1-carboxylic acid (ACC). By using an optimized targeted methodology, we rapidly isolated diverse soil- and plant-associated bacteria presenting phytohormone-degrading abilities from several plants, plant tissues and environments, without the need for prior extensive and laborious isolation and maintenance of large numbers of isolates. The developed methodology facilitates PGPB research, especially in developing countries. Here, we also report, for the first time, the isolation of bacterial strains able to concomitantly catabolize three phytohormones (SA, IAA and ACC). Ultimately, the described targeted methodology and the novel phytohormone-degrading bacteria obtained in this work may be useful tools for future plant-microbe interaction studies, and in the development of new inoculant formulations for agriculture and biotechnology.
Real-time PCR detection of a 16S rRNA single mutation of Helicobacter pylori isolates associated with reduced susceptibility and resistance to tetracycline in the gastroesophageal mucosa of individual hosts
The molecular mechanism of Helicobacter pylori resistance to tetracycline involves mutations in the primary binding site of the ribosome. A resistance or reduced susceptibility to tetracycline could be the result of single, double or triple mutations in the 16S rRNA gene of H. pylori . We investigated if the genotype was correlated to tetracycline resistance as determined phenotypically in vitro for 96 H . pylori isolates in the gastroesophageal mucosa of Venezuelan individual hosts. E-test for antimicrobial susceptibility test and real-time PCR for the detection of 16S rRNA gene mutations were performed in 96 H . pylori isolates (48 obtained from antrum, and 48 from oesophagus) from eight dyspeptic patients. In the gastric mucosa, 38 isolates were identified sensitive and 10 resistant to tetracycline by E-test, whereas 44 sensitive and 4 resistant isolates were found in the oesophagus. Real-time PCR detection of the 16S rRNA gene exhibited mutants with a single base-pair substitution (AGA926 GGA) in six antrum isolates and seven oesophagus isolates, whereas only three harboured a low level of tetracycline resistance in vitro. Our results indicate that real-time PCR detection of 16S rRNA is a reliable method to classify among tetracycline-resistant genotypes and useful in patients who have experienced a first-line treatment failure with triple therapy.
In vitro activities of six antifungal agents and their combinations against Chaetomium spp.Lingyue Sun, Zhe Wan, Ruoyu Li and Jin Yu
Purpose. To assess in vitro activities of six antifungal agents (amphotericin B, itraconazole, voriconazole, posaconazole, caspofungin and terbinafine) and the combined effects of eight pairs of them (caspofungin or terbinafine with amphotericin B, itraconazole, voriconazole or posaconazole) against 22 isolates of Chaetomium spp.
Methodology. The broth microdilution method drafted by the Clinical and Laboratory Standards Institute and the checkerboard method were used in this study to evaluate in vitro activities of antifungal drugs both alone and in combination against Chaetomium spp.
Results. Amphotericin B and triazoles exhibited lower geometric mean, MIC50 and MIC90 than caspofungin and terbinafine. Besides, all the paired drugs displayed varying degrees of synergism, with the interactions between caspofungin and itraconazole ranking first (86.36 %).
Conclusion. Our study illustrated varying degrees of synergism between caspofungin or terbinafine and itraconazole, voriconazole, posaconazole or amphotericin B towards Chaetomium spp., which could be a reference for the clinical treatment of Chaetomium spp. infections.
In vitro activity of mecillinam and nitroxoline against Neisseria gonorrhoeae – re-purposing old antibiotics in the multi-drug resistance era
In 2018, the European Centre for Disease Prevention and Control reported the first cases of extensively drug-resistant Neisseria gonorrhoeae infections in Europe. Seeking new options for antimicrobial therapy we investigated the susceptibility of N. gonorrhoeae to nitroxoline (NIT) and mecillinam (MCM), both of which are currently only indicated to treat uncomplicated urinary tract infections. Clinical N. gonorrhoeae isolates with non-susceptibility to penicillin from two German medical centres were included (n =27). Most isolates were also non-susceptible to a range of other anti-gonococcal antimicrobials (cefotaxime, ciprofloxacin, azithromycin, tetracycline). All isolates were further characterized by multi-locus sequence typing. MICs of penicillin and cefotaxime were determined by agar gradient diffusion. Production of penicillinase was tested by cefinase disk test. Susceptibility of MCM was investigated by agar dilution, NIT by agar dilution and disk diffusion. Penicillin MICs ranged from 0.125 to 64 mg l−1 and MICs of cefotaxime ranged from < 0.016 to 1 mg l−1 . Five isolates were penicillinase-producers. MICs of MCM ranged from 16 to > 128 mg l−1 whereas MICs of NIT ranged from 0.125 to 2 mg l−1 . NIT disk diffusion (median zone diameter 32 mm) correlated well with results from agar dilution. We demonstrated excellent in vitro activity of NIT against clinical N. gonorrhoeae isolates with non-susceptibility to standard anti-gonococcal antibiotics. MCM activity was unsatisfactory. Correlation of agar dilution and disk diffusion in NIT susceptibility testing is an important aspect with potential clinical implications.
In vitro efficacy of sodium selenite in reducing toxin production, spore outgrowth and antibiotic resistance in hypervirulent Clostridium difficile
Purpose. This study investigated the efficacy of the essential mineral, selenium (sodium selenite), in reducing the toxin production, spore outgrowth and antibiotic resistance of Clostridium difficile in vitro.
Methodology. Two hypervirulent C. difficile isolates were cultured in brain heart infusion broth with and without a sub-minimum inhibitory concentration (sub-MIC) of sodium selenite, and the supernatant and bacterial pellet were harvested for total toxin quantitation and RT-qPCR analysis of toxin-encoding genes, respectively. Additionally, C. difficile isolates were cultured in brain heart infusion broth containing 0.5 or 1× the minimum inhibitory concentration (MIC) of either ciprofloxacin or vancomycin with or without sub-MICs of sodium selenite. Further, the effect of sodium selenite on C. difficile germination and spore outgrowth was also determined by exposing C. difficile spores to a sub-MIC of sodium selenite in a germination medium and measuring the germination and outgrowth by measuring the optical density at 600 nm.
Results. Sodium selenite significantly reduced C. difficile toxin synthesis, cytotoxicity and spore outgrowth. Further, the expression of the toxin production genes, tcdA and tcdB, was downregulated in the presence of sodium selenite, while sodium selenite significantly increased the sensitivity of C. difficile to ciprofloxacin , but not vancomycin, as revealed by decreased bacterial growth in samples containing ciprofloxacin+selenium compared to the antibiotic control. Although the sub-MIC of sodium selenite did not inhibit spore germination, it was capable of completely inhibiting spore outgrowth.
Conclusion. Our results suggest that sodium selenite could potentially be used to control C. difficile and indicate that future in vivo studies are warranted.
Microbiome digital signature of MCR genes – an in silico approach to study the diversity of methanogenic population in laboratory-developed and pilot-scale anaerobic digesters
The production of biogas by anaerobic digestion (AD) of organic/biological wastes has a firm place in sustainable energy production. A simple and cost-effective anaerobic jar at a laboratory scale is a prerequisite to study the microbial community involved in biomass conversion and releasing of methane gas. In this study, a simulation was carried out using a laboratory-modified anaerobic-jar-converted digester (AD1) with that of a commercial/pilot-scale anaerobic digester (AD2). Taxonomic profiling of biogas-producing communities by means of high-throughput methyl coenzyme-M reductase α-subunit (mcrA) gene amplicon sequencing provided high-resolution insights into bacterial and archaeal structures of AD assemblages and their linkages to fed substrates and process parameters. Commonly, the bacterial phyla Euryarchaeota , Chordata, Firmicutes and Proteobacteria appeared to dominate biogas communities in varying abundances depending on the apparent process conditions. Key micro-organisms identified from AD were Methanocorpusculum labreanum and Methanobacterium formicicum . Specific biogas production was found to be significantly correlating to Methanosarcinaceae . It can be implied from this study that the metagenomic sequencing data was able to dissect the microbial community structure in the digesters. The data gathered indicates that the anaerobic-jar system could throw light on the population dynamics of the methanogens at laboratory scale and its effectiveness at large-scale production of bio-methane. The genome sequence information of non-cultivable biogas community members, metagenome sequencing including assembly and binning strategies will be highly valuable in determining the efficacy of an anaerobic digester.
Rapid antimicrobial susceptibility tests for sepsis; the road aheadMore Less
Current methods for antimicrobial susceptibility testing (AST) are too slow to affect initial treatment decisions in the early stages of sepsis, when the prescriber is most concerned to select effective therapy immediately, rather than finding out what will not work 1 or 2 days later. There is a clear need for much faster differentiation between viral and bacterial infection, and AST, linked to earlier aetiological diagnosis, without sacrificing either the accuracy of quantitative AST or the low cost of qualitative AST. Truly rapid AST methods are eagerly awaited, and there are several candidate technologies that aim to improve the targeting of our limited stock of effective antimicrobial agents. However, none of these technologies are approaching the point of care and nor can they be described as truly culture-independent diagnostic tests. Rapid chemical and genomic methods of resistance detection are not yet reliable predictors of antimicrobial susceptibility and often rely on prior bacterial isolation. In order to resolve the trade-off between diagnostic confidence and therapeutic efficacy in increasingly antimicrobial-resistant sepsis, we propose a series of three linked decision milestones: initial clinical assessment (e.g. qSOFA score) within 10 min, initial laboratory tests and presumptive antimicrobial therapy within 1 h, and definitive AST with corresponding antimicrobial amendment within an 8 h window (i.e. the same working day). Truly rapid AST methods therefore must be integrated into the clinical laboratory workflow to ensure maximum impact on clinical outcomes of sepsis, and diagnostic and antimicrobial stewardship. The requisite series of development stages come with a substantial regulatory burden that hinders the translation of innovation into practice. The regulatory hurdles for the adoption of rapid AST technology emphasize technical accuracy, but progress will also rely on the effect rapid AST has on prescribing behaviour by physicians managing the care of patients with sepsis. Early adopters in well-equipped teaching centres in close proximity to large clinical laboratories are likely to be early beneficiaries of rapid AST, while simplified and lower-cost technology is needed to support poorly resourced hospitals in developing countries, with their higher burden of AMR. If we really want the clinical laboratory to deliver a specific, same-day diagnosis underpinned by definitive AST results, we are going to have to advocate more effectively for the clinical benefits of bacterial detection and susceptibility testing at critical decision points in the sepsis management pathway.
Semisynthetic eugenol derivatives as antifungal agents against dermatophytes of the genus Trichophyton
Purpose. Eugenol, the main component of clove bud essential oil (Eugenia caryophyllus), has been linked to antimicrobial, anti-inflammatory, insecticidal and immunomodulatory properties. The purpose of this study was to evaluate the antifungal and cytotoxic activity of eugenol, the essential oil of Eugenia caryophyllus, and some semisynthetic derivatives of eugenol against dermatophytes of the genus Trichophyton.
Methodology. We evaluated the antifungal effect of the compounds, determining the minimum inhibitory concentrations (MICs) by the microdilution method and the minimum fungicidal concentrations by cultures from the inhibitions. Additionally, the inhibition of the radial growth of the mycelium of the dermatophyte fungi was tested by poisoned substrate. Cytotoxicity was measured by the colorimetric method on Vero cells.
Results. All of the eugenol compounds tested exhibited antifungal properties, showing MICs of 62.5–500 µg ml−1 , determined within three dermatophyte species: Trichophyton rubrum, Trichophyton mentagrophytes and Trichophyton tonsurans. Among these derivatives, methyl isoeugenol, at concentrations of 300 and 100 µg ml−1, was found to completely inhibit (100 %) radial growth of the mycelium of all three species after 20 days of treatment. Additionally, phenotypic variations related to the decrease in pigment production of T. rubrum were observed after treatment with O-ethyl and O-butyl isoeugenol derivatives. Meanwhile, all of the tested (iso)eugenol molecules exhibited moderate toxicity in Vero cells [50 % cytotoxic concentration (the concentration required for a 50 % reduction in cell viability; CC50): 54.06–265.18 µg ml−1 ).
Conclusion. The results suggest that the semisynthetic eugenol derivatives (SEDs) show promising antifungal activity and selectivity against dermatophyte fungi.
Casing microbiome dynamics during button mushroom cultivation: implications for dry and wet bubble diseases
The casing material required in mushroom cultivation presents a very rich ecological niche, which is inhabited by a diverse population of bacteria and fungi. In this work three different casing materials, blonde peat, black peat and a 50 : 50 mixture of both, were compared for their capacity to show a natural suppressive response against dry bubble, Lecanicillium fungicola (Preuss) Zare and Gams, and wet bubble, Mycogone perniciosa (Magnus) Delacroix. The highest mushroom production was collected from crops cultivated using the mixed casing and black peat, which were not significantly different in yield. However, artificial infection with mycoparasites resulted in similar yield losses irrespective of the material used, indicating that the casing materials do not confer advantages in disease suppression. The composition of the microbiome of the 50 : 50 casing mixture along the crop cycle and the compost and basidiomes was evaluated through next-generation sequencing (NGS) of the V3–V4 region of the bacterial 16S rRNA gene and the fungal ITS2 region. Once colonized by Agaricus bisporus, the bacterial diversity of the casing microbiome increased and the fungal diversity drastically decreased. From then on, the composition of the casing microbiome remained relatively stable. Analysis of the composition of the bacterial microbiome in basidiomes indicated that it is highly influenced by the casing microbiota. Notably, L. fungicola was consistently detected in uninoculated control samples of compost and casing using NGS, even in asymptomatic crops. This suggests that the naturally established casing microbiota was able to help to suppress disease development when inoculum levels were low, but was not effective in suppressing high pressure from artificially introduced fungal inoculum. Determination of the composition of the casing microbiome paves the way for the development of synthetic casing communities that can be used to investigate the role of specific components of the casing microbiota in mushroom production and disease control.
Alginate genes are required for optimal soil colonization and persistence by Pseudomonas fluorescens Pf0-1More Less
Pseudomonas fluorescens strains are important candidates for use as biological control agents to reduce fungal diseases on crop plants. To understand the ecological success of these bacteria and for successful and stable biological control, determination of how these bacteria colonize and persist in soil environments is critical. Here we show that P. fluorescens Pf0-1 is negatively impacted by reduced water availability in soil, but adapts and persists. A pilot transcriptomic study of Pf0-1 colonizing moist and dehydrated soil was used to identify candidate genetic loci, which could play a role in the adaptation to dehydration. Genes predicted to specify alginate production were identified and chosen for functional evaluation. Using deletion mutants, predicted alginate biosynthesis genes were shown to be important for optimal colonization of moist soil, and necessary for adaptation to reduced water availability in dried soil. Our findings extend in vitro studies of water stress into a more natural system and suggest alginate may be an essential extracellular product for the lifestyle of P. fluorescens when growing in soil.
Caribbean multi-centre study of Klebsiella pneumoniae: whole-genome sequencing, antimicrobial resistance and virulence factorsMore Less
The surveillance of antimicrobial-resistant isolates has proven to be one of the most valuable tools to understand the global rise of multidrug-resistant bacterial pathogens. We report the first insights into the current situation in the Caribbean, where a pilot project to monitor antimicrobial resistance (AMR) through phenotypic resistance measurements combined with whole-genome sequencing was set up in collaboration with the Caribbean Public Health Agency (CARPHA). Our first study focused on Klebsiella pneumoniae , a highly relevant organism amongst the Gram-negative opportunistic pathogens worldwide causing hospital- and community-acquired infections. Our results show that not only carbapenem resistance, but also hypervirulent strains, are circulating in patients in the Caribbean. Our current data does not allow us to infer their prevalence in the population. We argue for the urgent need to further support AMR surveillance and stewardship in this almost uncharted territory, which can make a significant impact on the reduction of antimicrobial usage. This article contains data hosted by Microreact (https://microreact.org).
Genomic surveillance of Escherichia coli in municipal wastewater treatment plants as an indicator of clinically relevant pathogens and their resistance genes
We examined whether genomic surveillance of Escherichia coli in wastewater could capture the dominant E. coli lineages associated with bloodstream infection and livestock in the East of England, together with the antibiotic-resistance genes circulating in the wider E. coli population. Treated and untreated wastewater was taken from 20 municipal treatment plants in the East of England, half in direct receipt of acute hospital waste. All samples were culture positive for E. coli , and all but one were positive for extended-spectrum β-lactamase (ESBL)-producing E. coli . The most stringent wastewater treatment (tertiary including UV light) did not eradicate ESBL- E. coli in 2/3 cases. We sequenced 388 E. coli (192 ESBL, 196 non-ESBL). Multilocus sequence type (ST) diversity was similar between plants in direct receipt of hospital waste versus the remainder (93 vs 95 STs, respectively). We compared the genomes of wastewater E. coli with isolates from bloodstream infection (n=437), and livestock farms and retail meat (n=431) in the East of England. A total of 19/20 wastewater plants contained one or more of the three most common STs associated with bloodstream infection (ST131, ST73, ST95), and 14/20 contained the most common livestock ST (ST10). In an analysis of 1254 genomes (2 cryptic E. coli were excluded), wastewater isolates were distributed across the phylogeny and intermixed with isolates from humans and livestock. Ten bla CTX-M elements were identified in E. coli isolated from wastewater, together with a further 47 genes encoding resistance to the major antibiotic drug groups. Genes encoding resistance to colistin and the carbapenems were not detected. Genomic surveillance of E. coli in wastewater could be used to monitor new and circulating lineages and resistance determinants of public-health importance.
Molecular characterization and antifungal susceptibility testing of Candida nivariensis from blood samples – an Iranian multicentre study and a review of the literature
Purpose. Identification of the emerging yeast species Candida nivariensis among presumptively identified Iranian Candida glabrata isolates.
Methodology. Clinical C. glabrata species complex isolates from blood (n=100; 46.9%), vaginal swabs (n=20; 9.4%), bronchoalveolar lavage (n=10; 4.7%) and sputum (n=12; 5.6%) from 68 patients from Iran were investigated. Isolates were characterized by CHROMagar, multiplex PCRs, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), amplified fragment length polymorphism (AFLP) fingerprinting, internal transcribed spacer (ITS)/large subunit (LSU) rDNA and FKS1/FKS2 sequencing, and the European Committee on Antimicrobial Susceptibility Testing broth microdilution method. A comprehensive literature review was conducted and all the relevant clinical and microbiological data were collected.
Results. Four C. nivariensis isolates were recovered from blood samples of three subjects and were all consistently identified by nine-plex PCR, Bruker MALDI-TOF MS, and LSU and ITS rDNA sequencing. AFLP genotyping clustered the isolates into two groups. Sequencing of the FKS1 and FKS2 hotspots showed no accountable amino acid substitutions. All isolates were susceptible to amphotericin B, fluconazole, itraconazole, posaconazole, voriconazole, anidulafungin and micafungin.
Conclusion. In total, 4 out of 213 clinical C. glabrata species complex candidemia isolates were C. nivariensis. Improvement of the BioMerieux Vitek MS database is required to accurately identify C. nivariensis and it is advised to alternatively use CHROMagar and/or PCR-based techniques. As other species within the Nakaseomyces clade may cause infection and showed high MIC values for antifungals, inclusion of their spectra into the MALDI-TOF MS database seems relevant. Due to developing resistance to fluconazole and insufficient efficacy of caspofungin, the combination of catheter removal plus treatment with caspofungin, or voriconazole, or micafungin might be effective for patients.
Streptomyces fodineus sp. nov., an actinobacterium with antifungal activity isolated from mine area soilMore Less
A novel actinobacterial strain producing an antifungal substance was isolated from a sample of acidic mine area soil, and its taxonomic position was evaluated. The novel strain, designated TW1S1T, formed white-grey aerial mycelium and yellow substrate mycelium on oatmeal agar. Growth occurred at 10–45 °C (optimum, 30 °C), pH 4–9 (pH 6–7) and in the presence of up to 8 % (w/v) NaCl. Melanin was produced on peptone–yeast extract–iron agar. Phylogenetic analysis based on its 16S rRNA gene sequence indicated that the novel strain should be assigned to the genus Streptomyces , and the closest species was Streptomyces puniciscabiei S77T with 99.1 % sequence similarity, which was followed by Streptomyces durhamensis NRRL B-3309T (99.0 %), Streptomyces filipinensis NBRC 12860T (98.9 %) and Streptomyces yaanensis Z4T (98.7 %). The chemotaxonomic properties were consistent with those of Streptomyces . ll-Diaminopimelic acid was the diagnostic diamino acid, and alanine, glutamic acid and glycine were present in the peptidoglycan. The cell-wall hydrolysate also contained galactose, glucose, mannose and ribose. The predominant isoprenoid quinones were MK-9(H4) and MK-9(H6), the major polar lipids were phosphatidylglycerol and an unidentified phospholipid, and the main fatty acids were iso-C16 : 0 and anteiso-C15 : 0. However, strain TW1S1T could be distinguished from its neighbouring species by its phenotypic properties. In addition, the genome-based comparison with the closest species indicated that strain TW1S1T should be recognized as a separate species. The phylogenetic, phenotypic and chemotaxonomic as well as genomic evidence supported that TW1S1T represents a novel species of Streptomyces , for which the name Streptomyces fodineus sp. nov. is proposed (type strain, TW1S1T = KCTC 49013T = JCM 32404T).
Use of whole genome sequencing in surveillance for antimicrobial-resistant Shigella sonnei infections acquired from domestic and international sourcesMore Less
Shigella species are a major cause of gastroenteritis worldwide, and Shigella sonnei is the most common species isolated within the United States. Previous surveillance work in Pennsylvania documented increased antimicrobial resistance (AMR) in S. sonnei associated with reported illnesses. The present study examined a subset of these isolates by whole genome sequencing (WGS) to determine the relationship between domestic and international isolates, to identify genes that may be useful for identifying specific Global Lineages of S. sonnei and to test the accuracy of WGS for predicting AMR phenotype. A collection of 22 antimicrobial-resistant isolates from patients infected within the United States or while travelling internationally between 2009 and 2014 was chosen for WGS. Phylogenetic analysis revealed both international and domestic isolates were one of two previously defined Global Lineages of S. sonnei , designated Lineage II and Lineage III. Twelve of 17 alleles tested distinguish these two lineages. Lastly, genome analysis was used to identify AMR determinants. Genotypic analysis was concordant with phenotypic resistance for six of eight antibiotic classes. For aminoglycosides and trimethoprim, resistance genes were identified in two and three phenotypically sensitive isolates, respectively. This article contains data hosted by Microreact.
Antifungal susceptibilities, biofilms, phospholipase and proteinase activities in the Candida rugosa complex and Candida pararugosa isolated from tertiary teaching hospitals
Purpose. Non-albicans Candida species have emerged as fungal pathogens that cause invasive infections, with many of these species displaying resistance to commonly used antifungal agents. This study was confined to studying the characteristics of clinical isolates of the C. rugosa complex and C. pararugosa species.
Methodology. Seven isolates of the C. rugosa complex and one isolate of C. pararugosa were obtained from two tertiary referral hospitals in Malaysia. Their antifungal susceptibilities, biofilm, proteinase, phospholipase, esterase and haemolysin activities were characterized. Biofilms were quantified using crystal violet (CV) and tetrazolium (XTT) reduction assays at 1.5, 6, 18, 24, 48 and 72 h.
Results/Key findings. The E-test antifungal tests showed that both species have elevated MICs compared to C. albicans and C. tropicalis. The highest biomass was observed in one of the C. rugosa isolates (0.237), followed by C. pararugosa (0.206) at 18 h of incubation. However, the highest bioactivity was observed in the C. rugosa ATCC 10571 strain at 24 h (0.075), followed by C. pararugosa at 48 h (0.048) and the same C. rugosa strain at 24 h (0.046), with P<0.05. All isolates exhibited high proteinase activity (+++) whereas six isolates showed very strong esterase activity (++++). All the isolates were alpha haemolytic producers. None of the isolates exhibited phospholipase activity.
Conclusion. Elevated MICs were shown for the C. rugosa complex and C. pararugosa for commonly used antifungal drugs. Further studies to identify virulence genes involved in the pathogenesis and genes that confer reduced drug susceptibility in these species are proposed.
Kodoja: A workflow for virus detection in plants using k-mer analysis of RNA-sequencing dataMore Less
RNA-sequencing of plant material allows for hypothesis-free detection of multiple viruses simultaneously. This methodology relies on bioinformatics workflows for virus identification. Most workflows are designed for human clinical data, and few go beyond sequence mapping for virus identification. We present a new workflow (Kodoja) for the detection of plant virus sequences in RNA-sequence data. Kodoja uses k-mer profiling at the nucleotide level and sequence mapping at the protein level by integrating two existing tools Kraken and Kaiju. Kodoja was tested on three existing RNA-seq datasets from grapevine, and two new RNA-seq datasets from raspberry. For grapevine, Kodoja was shown to be more sensitive than a method based on contig building and blast alignments (27 viruses detected compared to 19). The application of Kodoja to raspberry, showed that field-grown raspberries were infected by multiple viruses, and that RNA-seq can identify lower amounts of virus material than reverse transcriptase PCR. This work enabled the design of new PCR-primers for detection of Raspberry yellow net virus and Beet ringspot virus. Kodoja is a sensitive method for plant virus discovery in field samples and enables the design of more accurate primers for detection. Kodoja is available to install through Bioconda and as a tool within Galaxy.


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What Happens to Those Who Never Hear the Gospel?
by Matt Smethurst
The man on the island. Perhaps you've encountered him in a friend's argument against Christianity. Maybe you've even voiced the objection yourself.
How could a good and loving God condemn to hell someone who's never heard of him?
When it comes to this emotionally vexing issue, there are two dominant positions among professing Christians: inclusivism and exclusivism. While both views maintain that Jesus is the only way to God, only one insists on the necessity of conscious faith in him.
Inclusivism vs. Exclusivism
Read the entire article here.
HERMENEUTICS: Analogia Scripturae and Analogia Fidei
By Bill Hier
This is the title of chapter two of CONFESSING THE IMPASSIBLE GOD: The Biblical, Classical, & Confessional Doctrine of Divine Impassibility (CIG). The purpose of this chapter is to highlight the two most necessary hermeneutical principles that are required when doing theology – not only theology proper, as is the concern of CIG, but all theology. As the title states, these are the hermeneutical principles of Analogia Scripturae and Analogia Fidei, which are the Latin phrases for The Analogy of Scripture and The Analogy of the Faith.
Before going forward, defining these most important hermeneutical principles, and stating where they come from, is necessary.
To put it simply, these principles are not formulated and then imposed upon Scripture, but rather, and drawn from the way that the Biblical writers themselves did theology. Thus, they come from Scripture, and so, from God – they are principles of understanding Scripture which the Author of Scripture imbedded in His Special Revelation to us, that we might not make the mistake of pitting Scripture against Scripture, but could rather understand it, and all the doctrines which it teaches us, by a synthesis of the whole.
Read the entire article here.
Bell's Hell and the Destiny of Those Who've Never Heard of Jesus
by Sam Storm
In a recent interview with Sally Quinn of The Washington Post, Rob Bell again muddied the waters over the question of the fate of those who've never heard about Jesus. In doing so he also greatly misrepresented the evangelical answer to this question. Here are his words:
"If, billions and billions and billions of people, God is going to torture them in hell forever – people who never heard about Jesus are going to suffer in eternal agony because they didn't believe in the Jesus they never heard of – then at that point we will have far bigger problems than a book from a pastor from Grand Rapids."
Bell is responding to evangelicals who purportedly believe that people "are going to suffer in eternal agony because they didn't believe in the Jesus they never heard of." Let me say this as clearly as I can: No one will ever suffer for any length of time in hell or anywhere else for not believing in the Jesus they never heard of. Should I say that again or is it enough to ask that you go back and read it again?
Bell and others who make this sort of outrageous claim have evidently failed to look closely at Romans 1:18ff. Here we read that the wrath of God revealed from heaven is grounded in the persistent repudiation by mankind of the revelation God has made of himself in the created order. In other words, there is a reason for God's wrath. It is not capricious. God's wrath has been deliberately and persistently provoked by man's willful rejection of God as he has revealed himself.
Read the entire article here.
Also check out: "What About Those Who have Never Heard of Jesus?A Classic Illustration From Francis Schaeffer"
God's Eternality and Self Existence
The attributes of God as described by Moses, David, and Jeremiah. Explanation of the attributes. Summary. Uses of this knowledge.
2. There are certain passages which contain more vivid descriptions of the divine character, setting it before us as if his genuine countenance were visibly portrayed. Moses, indeed, seems to have intended briefly to comprehend whatever may be known of God by man, when he said, "The Lord, The Lord God, merciful and gracious, long-suffering, and abundant in goodness and truth, keeping mercy for thousands, forgiving iniquity and transgression and sin, and that will by no means clear the guilty; visiting the iniquity of the fathers upon the children, and upon the children's children, unto the third and to the fourth generation," (Ex. 34:6, 7.) Here we may observe, firsts that his eternity and self existence are declared by his magnificent name twice repeated; and, secondly, that in the enumeration of his perfections, he is described not as he is in himself, but in relation to us, in order that our acknowledgment of him may be more a vivid actual impression than empty visionary speculation. Moreover, the perfections thus enumerated are just those which we saw shining in the heavens, and on the earth — compassion, goodness, mercy, justice, judgment, and truth. For power and energy are comprehended under the name Jehovah. Similar epithets are employed by the prophets when they would fully declare his sacred name. Not to collect a great number of passages, it may suffice at present to refer to one Psalm, (145) in which a summary of the divine perfections is so carefully given that not one seems to have been omitted. Still, however, every perfection there set down may be contemplated in creation; and, hence, such as we feel him to be when experience is our guide, such he declares himself to be by his word. In Jeremiah, where God proclaims the character in which he would have us to acknowledge him, though the description is not so full, it is substantially the same. "Let him that glorieth," says he, "glory in this, that he understandeth and knoweth me, that I am the Lord which exercise loving-kindness, judgment, and righteousness, in the earth," (Jeremiah 9:24.) Assuredly, the attributes which it is most necessary for us to know are these three: Loving-kindness, on which alone our entire safety depends: Judgment, which is daily exercised on the wicked, and awaits them in a severer form, even for eternal destruction: Righteousness, by which the faithful are preserved, and most benignly cherished. The prophet declares, that when you understand these, you are amply furnished with the means of glorying in God. Nor is there here any omission of his truth, or power, or holiness, or goodness. For how could this knowledge of his loving-kindness, judgment, and righteousness, exist, if it were not founded on his inviolable truth? How, again, could it be believed that he governs the earth with judgment and righteousness, without presupposing his mighty power? Whence, too, his loving-kindness, but from his goodness? In fine, if all his ways are loving-kindness, judgment, and righteousness, his holiness also is thereby conspicuous. Moreover, the knowledge of God, which is set before us in the Scriptures, is designed for the same purpose as that which shines in creation, viz., that we may thereby learn to worship him with perfect integrity of heart and unfeigned obedience, and also to depend entirely on his goodness.
John Calvin-Institutes of the Christian Religion-Book I-Chapter 10-Henry Beveridge Translation
Scripture represents God's character as the same as revealed through his works
Explanation of the knowledge of God resumed. God as manifested in Scripture, the same as delineated in his works.
1. We formerly observed that the knowledge of God, which, in other respects, is not obscurely exhibited in the frame of the world, and in all the creatures, is more clearly and familiarly explained by the word. It may now be proper to show, that in Scripture the Lord represents himself in the same character in which we have already seen that he is delineated in his works. A full discussion of this subject would occupy a large space. But it will here be sufficient to furnish a kind of index, by attending to which the pious reader may be enabled to understand what knowledge of God he ought chiefly to search for in Scripture, and be directed as to the mode of conducting the search. I am not now adverting to the peculiar covenant by which God distinguished the race of Abraham from the rest of the nations. For when by gratuitous adoption he admitted those who were enemies to the rank of sons, he even then acted in the character of a Redeemer. At present, however, we are employed in considering that knowledge which stops short at the creation of the world, without ascending to Christ the Mediator. But though it will soon be necessary to quote certain passages from the New Testament, (proofs being there given both of the power of God the Creator, and of his providence in the preservation of what he originally created,) I wish the reader to remember what my present purpose is, that he may not wander from the proper subject. Briefly, then, it will be sufficient for him at present to understand how God, the Creator of heaven and earth, governs the world which was made by him. In every part of Scripture we meet with descriptions of his paternal kindness and readiness to do good, and we also meet with examples of severity which show that he is the just punisher of the wicked, especially when they continue obstinate notwithstanding of all his forbearance.
John Calvin-Institutes of the Christian Religion-Book I-Chapter 10-Henry Beveridge Translation
Chapter 3-Reason and Revelation
Chapter 3-Reason and Revelation
HAVING considered the proofs of the existence of God, we should discuss the ways in which he has made himself known, before we study his nature, and attributes, and relations to us. These constitute the sources of our knowledge of Theology, which are two, Reason and Revelation.
Reason is that power in man, which enables him to have mental perceptions, to exercise thought, and reflection, to know facts, to inquire into their mutual relations, and to deduce, logically, the conclusions which may be drawn from them.
Reason may be used either with reference to the natural or supernatural means of knowledge conferred by God.
When we refer to reason as a source of knowledge distinct from revelation, we mean the information attained, by the use of this faculty, in connection only with the natural, as distinguished from the supernatural.
By revelation, we mean the knowledge which God conveys by direct supernatural instruction, pre-eminently that given in the book known as the Bible.
Reason involves all the cognitive powers of man, which are the faculties through which the mind attains knowledge. These faculties are not separate, and independent, but are merely the instruments of the mind.
The mind is not itself an original source of knowledge, like the Scriptures, but is merely an instrument by which the man attains knowledge through the exercise of its appropriate faculties. There are no such things as innate ideas. These arrive only through the exercise of proper thought and reflection, in connection with some perceived facts.
The means by which the mind attains knowledge in the exercise of its faculties, are five.
1. Consciousness, by which we learn our own existence, and the fact that we think, and are personal beings, possessing personal identity during the term of our natural life.
2. Observation, and experience of the world about us, through the senses.
3. Through intuitive conceptions, by which, upon the suggestion through some external object, of some principle, we find ourselves at once convinced of its correctness.
4. The dispositions, instincts and tendencies of our natures.
5. The curse of events in nature, as tending to good or evil, to what is desirable or disastrous.
It is manifest that the knowledge obtained from these various sources must be abundant to teach man the simple facts upon which rests his duty to God; namely, that there is a God to whom he owes existence, and consequent reverence, service and love, and whose greatness and goodness enforce this obligation; also to show him that that duty has not been discharged, and that he has not the disposition to discharge it; and consequently to render him uneasy in his relations to God, and anxious to appease him, and secure some assurance of his pardon and approval. It has also been thought by many, that through reason alone man attains the conviction of immortality and of a future state of rewards and punishments.
However abundant may be the information thus conveyed to man, it is nevertheless clear that his knowledge in these directions must still remain very imperfect.
This must have been true of man even in a state of innocence. His finite nature and the finite conditions which surrounded him must still have left him ignorant upon many desirable matters. It is natural, therefore, to believe that, in that condition, he received direct communications from God, which are properly esteemed revelations.
But this imperfection must have been greatly increased by an subsequent, fall from innocence. By this the Perceptions of right and wrong would be dimmed, the power of conscience to enforce the right would be impaired, the desire to do the right would be diminished, prejudices against the right would be created, an affection for God would be greatly decreased, if not entirely obliterated.
Upon these grounds we may infer the necessity of some further source of knowledge of God, and of his will with respect to man.
We may also argue a priori as to the nature of this revelation.
1. It must come from God, the source of all our other knowledge. No other could give it, and it is fit that no other should do so.
2. It must be suited to our present condition, confirming the truth already known, and teaching what is practically useful to man as sinner before God.
3. It must be secured from all possibility of error, so that its teachings may be relied on with equal, if not greater, confidence than those of reason.
4. It must come with authority, claiming and proving its claim to be the word of God, who has the right to command, and to punish those who disobey his commands; with authority also, that man may with confidence believe and trust the promises and hopes pardon and peace it may hold out.
5 That it will be accompanied by difficulties and mysteries what may be expected, since these are found frequently attending the knowledge derived from reason.
The gift of such a revelation must of course depend absolutely upon the will of God. It is not for man to say, before it is given, whether it certainly will, or will not, be bestowed.
That it is not improbable may be inferred from the fact that God has already made himself known to us in various ways in ourselves and in nature. If we need further revelation we my hope for it.
The only reason to the contrary is that we have sinned against God, and he may have chosen to abandon us to our fate. But this is not so truly understood until revelation has confirmed our conviction of our sinful estate. On the other hand, the favors which God still bestows, and the means of continued knowledge of him which he affords, indicate that he has not yet consigned us to our deserved fate, and that he may have purposes of mercy towards us.
That which renders it highly probable is the expectation seen in man, in the conceptions he has formed of God, as one to be propitiated by sacrifices and approached with prayer.
If the expectations thus formed are to he verified, the important question arises, in what way can God make known to us the new truth he wills to teach.
They manifestly speak unadvisedly who assert that this can in nowise be done.
If he should so choose, he could impress it on each one in like manner as we attain intuitive conceptions. He might reveal it to individuals in dreams and visions, so as to make each one feel and know that the vision is from God. Those through whom he has revealed himself have in some such way attained absolute conviction that God has spoken to and through them, and with God there is neither impossibility nor difficulty in producing like certainty in the mind of each individual of the race.
But as God usually acts through means, so he has revealed himself to a few, and through them to mankind in general.
The only question then is, how can he give evidence to the race at large that the men he has inspired are indeed his messengers?
This also might be done in various ways, but he has chosen to do it by attesting their mission by miracles wrought through them.
As to the measure of authority to be ascribed to these miracles, men differ in opinion.
Some teach that any miracle wrought is of itself sufficient attestation of the messenger and of the truth which he teaches.
Others, that miracles are only proofs to those who behold them, and dubious proofs even then, and that the true purpose of them is not to set the seal of God's authority, but simply to awaken attention and excite awe, and thus prepare the way for a proper hearing of the divine message. These assert that the revelation comes to us with the authority only of the self-convincing nature of the truth made known.
It is necessary, in this difference of opinions, to seek carefully after the true theory. From no source can we better obtain it than from the revelation itself, the teaching of which will be seen to be fully corroborated otherwise.
The Scripture theory seems to be this, that in any new revelation the prophet of God must present a doctrine perfectly consistent with ever past revelation and with the knowledge conveyed by nature, and must, at the same time, confirm by miracles his authority as a teacher from God. Without the miracle the new truth has no evidence that it is not simply the product of human reason or imagination. The coincidence in doctrine is necessary to protect against pretended miracles and the tricks of unprincipled men. Besides, the new truth can have no higher authority than the old, and therefore cannot supersede it, for the old also has come from God. No truth ever taught by God can be opposed by any new truth from him. What with God is truth is eternal truth. Like himself, it is the same "yesterday, to-day and forever." It may be more abundantly or clearly revealed. We may learn to comprehend it better and to correct our own misapprehensions of it, but whatever God has once given as truth must so remain forever, as changeless as his own life.
1. The Scriptural authority for this theory is conclusive.
Moses announced the law, which shows the miracle alone not to be conclusive. See Deut. 13:1, 2, 3. "If there arise in the midst of thee a prophet, or a dreamer of dreams, and he give thee a sign or a wonder, and the sign or the wonder come to pass, whereof he shake unto thee, saying, let us go after other gods which thou hast not known, and let us serve them; thou shalt not hearken unto the words of that prophet, or unto that dreamer of dreams: for the Lord your God proveth you to know whether ye love the Lord your God with all your heart and with all your soul." This passage shows that even a miracle, wrought by one teaching doctrine not in accordance with that already received, should not tempt to belief in the divine authority of him who should work it.
The Apostle Paul gives similar instruction to the Galatians, Gal. 1:8: " Though we, or an angel from heaven, should preach unto you any gospel other than that which we preached unto you, let him be anathema." Whatever might be the accredited authority of the messenger, his teachings were not to be received.
Yet, with all this, the Scriptures do not disparage the miracle. The miracles of Mosaic times are constantly referred to as indubitably marking it as divine. Nicodemus recognized the high position assigned to miracles by the Jews, John 3:2: "No man can do these signs that thou doest, except God be with him." Christ himself says, John 10:25: "The works that I do in my Father's name, these bear witness of me."
This theory of the Scriptures is not necessarily based upon the idea that real miracles can be wrought otherwise than by divine power. Still the language sometimes used is liable to this construction. And much depends upon the definition of a miracle. If a miracle be a suspension of the fixed laws which God has established for the world, that suspension can only occur through his special permission. Taking this as the true meaning of the word, we can understand why such stress is laid in the Scriptures upon the Mosaic miracles and those of Christ, since many of them are such as nothing but divine power could accomplish. But the word miracle in the Scriptures has not this restricted meaning, but is applied likewise to any marked supernatural event. Because men are apt to put these upon a level with the miracles which God alone can work, they are warned not to follow after what is thus supernaturally done, if it be accompanied by such teaching as is contrary to truth already received.
See the apparent reality of such miracles in connection with the magicians of Egypt, Ex. 7:11; Chap. 8:7, and compare with it the conviction expressed by the magicians, Ex. 8:19, when they failed to produce lice from the dust, "This is the finger of God."
Notice also what Christ says, Mark 13:22: "For there shall arise false Christs and false prophets, and shall shew signs and wonders, that they may lead astray, if possible, the elect."
See also Rev. 16:13, 14: "And I saw coming out of the mouth of the dragon, and out of the mouth of the beast, and out of the mouth of the false prophet, three unclean spirits, as it were frogs, for they are spirits of devils, working signs; which go forth unto the kings of the whole world, to gather them together unto the war of the great day of God the Almighty."
It is because of this liability to be deceived, that the Scriptures require the miracle and the concurrent doctrine as both essential to the reception of a new revelation.
2. This theory alone concurs with the course to which nature necessarily impels us.
To the extent that we are fully convinced of the truth of a doctrine, no subsequent revelation could change our belief. It is true that this does not apply when we have doubts; but when our knowledge is fixed, we cannot be moved. No amount of miracle could convince a Christian that the nature of God is otherwise than pure and holy, or that he delights in worship not of the heart, or that he is not infinite in justice and holiness, in goodness, mercy and truth, or that be will pardon sin without due satisfaction to his law.
3. This theory accords with the progressive character of divine revelation.
The earliest revelation came to those who had heretofore been guided only by reason. This was true even down to the beginnings of the Old Testament Scriptures, and, in that economy, only preparation was made for the future glory of the New Testament revelation. Hence the truths taught were, for the most part, only those which come within the compass of discovery by reason, or acceptance by it upon due suggestion, namely,–the existence of one God, the fact of creation, the law of moral obligation to God and man, the punishment of sinners, the duty of repentance, the pardoning mercy of God, and the law of sacrifices, with substitution and satisfaction.
The new economy goes further in its clear instructions: it teaches the vicarious atonement of Christ, involving representation in him and also in Adam, the doctrine of the Trinity in the Godhead, the mysterious union in the person of Christ, and many other truths heretofore only very indistinctly revealed.
These could not have been presented to those only taught heretofore by reason. But the revelation which stood between fore-shadowed them in different ways. From it alone originally they would not have been discovered. But now that they are made known, that former revelation is seen to concur with the new statements, and the conformity of the clearly expressed doctrine to the mere outlines of them in the past sustains the fact that they have a common author, and that the divine revealer is the same. It is like the presence in animals of the same genus in earlier days of germs which find their development in species which come later.
4. This accords with our means of judging what course of action infinite wisdom would have devised.
The conviction we have of past truth renders it impossible that we should throw it aside. We must, therefore, still hold it fast. That conviction has come from God, and we can have no higher evidence.
Yet, other statements and doctrines very probably or even certainly true, may be taught by men, as revealed to them, when they are either self-deceived, or attempting to deceive others. Hence, we must have the attesting miracle.
On the other hand, we are liable to be deceived as to what is supernatural, and especially, in the supernatural, as to what is within the limits of created power. Hence, we may be misled by the craft of men, or by the superhuman power of wicked spirits. Therefore, no doctrine must be accepted contrary to a truth already received.
A revelation, such as we have described, having been given and proved, another question arises: what is the relation which reason bears towards it?
We may lay down the following facts:
l. That reason is the first revelation, and is consequently presupposed in any other.
2. That the facts of reason cannot be denied by any subsequent revelation. No truth can destroy other truth.
A limitation must, however, be put on the province of reason. The doctrines of which it may judge, are those only which come within its sphere. Upon the presentation of a new doctrine reason may decide whether it agrees with former knowledge. If agreeable thereto, it must be accepted, if opposed, it must be rejected. But, if it be above reason, it must stand or fall with the rest of the revelation. God may, in his mercy, refrain from trying faith by a revelation of supernatural doctrine, but, if he reveals it, it must be no barrier to the reception of that doctrine itself, or of the revelation which accompanies it. In an able article in the Southern Presbyterian Review, Vol. I, pp. 1-34, on "Reason and Revelation," Dr. Thornwell puts this limitation upon reason, that it is sole arbiter within its own bounds, but no judge beyond them. He thinks that in this way only can it be applied as a test of doctrine. The theory is undoubtedly correct. It fails only in not recognizing the precise manner in which Scripture brings it in as an arbiter, not as the judge of truth as disconnected from the past, but as related to the various times and forms in which God has taught it. Reason should judge a new revelation, not by the truths taught by reason alone, but also by those which have been made known in any previous revelation.
The office of reason with respect to revelation, is therefore seen to be:
1. To examine the evidence of the miracles upon which it rests.
2. To compare its doctrines with the teaching of the past, and recognize their correspondence with or opposition to that teaching.
3. To adopt or reject the revelation according to the evidence afforded that it is God's truth.
4. To interpret its contents, according to the best light which learning affords.
Rev. James Petigru Boyce, D.D., LL. D.,–Abstract of Systematic Theology–First published in 1887
Free ebook-A Reformed Approach to Science and Scripture
A Reformed Approach to Science and Scripture, A New Free eBook from Keith Mathison
In A Reformed Approach to Science and Scripture, Dr. Mathison briefly introduces us to a topic that has long been a subject of debate, aiming to equip Christians with a clear foundation so that they may approach questions and discussions pertaining to science and Scripture with grace, humility, and patience.
Table of Contents
Foreword by R.C. Sproul
1. All Truth Is God's Truth
2. General and Special Revelation
3. Interpreting General and Special Revelation
4. Luther, Calvin, and Copernicus
5. Earthly Things and Heavenly Things
6. When Science and Scripture Conflict
7. The Age Of The Universe and Genesis 1
"Christians have absolutely nothing to fear ultimately from scientific research," says Dr. Mathison. "If scientists discover something about God's creation that is actually true, it will not and cannot ultimately contradict the Scriptures when they are properly interpreted."
Available Now as a Free Download


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By at January 30 2020 23:41:36
Here are some handy tips on how to go about writing the essay samples on Shakespeare. The dated language and the period settings of the story might make the plot intriguing for most students. Read the work thoroughly, take the aid of your teachers in clearing doubts or watch a movie version of the work, which will help you to grasp the essence of the story easily.
This is a particular type of writing essay that is often put for view by online research and writing services. As a student, you should not only consider looking at classification essay, you should also consider writing a sample essay that can be viewed as a sample paper by other students.
In most cases, students will turn to these essays because of the factor of time. Most students will keep research and writing till the last minutes before actually beginning the write-up. One of the most important aspects in any academic writing is the issue of time. In everything you do in academia, it is always necessary to make use of an outline. The outline guides you as you write from start to finish. The outline is there to ensure that you start and finish on time. Writing without an outline is what makes you get caught up by deadlines.
Well-researched argument, certainly. A considered viewpoint, developing themes within a structured, coherent essay, of course. Perhaps some original thinking, a breakthrough idea. All essays are different, naturally, but one common denominator of all great essays is clarity, flow, and consistency of presentation.


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Some frequently asked questions about Sodablast Systems
> Will the process damage the surface to be cleaned?
In contrast to sand blasting or bead blasting that works through abrasion, soda blasting does not damage the surface.
> Is sodablast better than sand or grit blasting?
Sodablast offers the advantage that it does not damage the treated surface. It does not give an impact on the abrasive. Sand and grit does. Blasted ferrous metals require an immediate coating to prevent corrosion. By contrast, sodablast will significantly reduce the speed of rust formation.
> Is sodablast good at cleaning engines and greasy parts?
Sodium bicarbonate breaks down hydrocarbons making it an excellent method for cleaning engines and / or greasy parts.
> Would sodablast help to remove graffiti?
When used with a little water, sodablast is a very good system to remove graffiti.
> What about small amounts of oil and grease on a surface to be cleaned?
Veel vet en olie vertragen het reinigingsproces maar soda zal niet achterblijven in vette hoeken in tegenstelling tot andere straalmiddelen .Tijdens het sodastralen zetten de onreinheden zich om in een zeepemulsie . Dit proces wordt verzeping genoemd.
> Is soda safe for the environment?
Soda has no influence on the environment. It is almost completely safe. This product is also used in cooking and also to combat stomach problems. Due to the alkaline properties, damage to plants and vegetation can occur if not enough rinsing is done with water.
> Is sodablast safe on rubber and plastics?
With sodablasting you can remove paint and other impurities in general without any problems.
> What about sound and dust
Some frequently asked questions


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What Do I Do If My Bicycle Arrives Damaged?
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Hey guys, Dustin here, CEO of six three zero. Want to talk to you today about what do you if my bike comes damaged.
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But that's how you submit it, just send an email, firstname.lastname@example.com, and detail what's going on. Please follow up with a photo and your order number, your order number as well.
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The Bipartisan Report would never call Donald Trump obese. That would be rude. It's the fat-shaming GOP candidate's body mass index (BMI) that's calling him obese.
As the old adage goes, fat shamers in glass penthouses shouldn't throw stones.
The alleged billionaire's many detractors had a field day with that ridiculous doctor's letter, which lauds his health as "extraordinary" and "astonishingly excellent." Just when you thought there was nothing left to pick over, a Daily Kos member came up with something.
The post by WestportDem hones in on a telling detail…Donald Trump likely lied about his weight (all that for just 236 pounds?) and definitely lied about his height. Even if those numbers are accurate, and they can't be, Trump is medically overweight. The truth suggests that he's clinically obese.
Dr. Harold Bornstein's letter, which the Trump-Pence campaign produced in lieu of actual medical records, says the GOP's White House hopeful is 6'3″ tall and weighs 236 pounds.
But Donald Trump's height is actually 6'2″.
So why on earth would Donald Trump lie about his height (and very likely his weight) when his height is actually a matter of public record? Seriously, if do a Google search for Trump's height, it comes up 6'2″.
Well…according to the Body Mass Index (BMI) chart below, being 6'3″ tall and weighing 236 pounds would put Donald Trump in the "overweight" category. That's slightly embarrassing, but he can always claim it's mostly muscle.
If Donald Trump turns out to be only 6'2″, then he falls solidly into the "obese" category. And Westport Dem found some photos that support this.
Here's the wanna-be Fat-Shamer-in-Chief standing next to A-Rod, who actually is 6'3 according to Google's search of records.
Melania Trump's 5'11" tall, but also looks like she's 6'2″ thanks to her high heels and pouffy coiffe.
Donald Trump lied about his weight, too.
As if we didn't already know Donald Trump lies about everything, the Daily Banter points out the wannabe Liar-in-Chief lied about his weight, too.
'There's no way to verify the numbers on Bornstein's letter, including Trump's weight, which was reported to be 236 pounds. However, earlier reports indicate Trump might actually weigh 267 pounds and, quite frankly, the heavier weight makes more sense given his 6-foot 3-inch frame. (Similarly, prior to Bornstein's latest stats for Trump, he was reported to be 6-foot-2, and not 6-foot-3. It's well within reason to suggest Trump might also be lying about his height.)'
Now, as many of the Bipartisan's smart readers will quickly point out, the Body Mass Index (BMI) is a blunt instrument. The Public Library of Science (PLOS) blog points out the BMI is a poor measurement of our weight and health. For starters, it doesn't account for muscle mass vs. fat mass. Nor does it account for weight distribution, since weight that concentrates in the stomach is more of a health risk than weight that accumulates more evenly. The BMI also doesn't account for lifestyle changes, like quitting smoking or exercising more.


text: This "married couple" (with no wedding rings) came out to meet with what they believed was a 13 year old girl after a sexual text conversation. These two individuals come across as human traffickers and were prowling in a area that is actually known for such activity. If you recognize either of these individuals do your part and use your voice !!


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We have received the following schedule for VITA Tax Assistance at the Mary H. Weir Public Library. Those providing this service are volunteers who are sharing their time and knowledge to help the community. The schedule may change if necessary.
Tuesday, December 10, 2019
VITA Tax Assistance
Posted by Your Friendly Neighborhood Librarian at 10:14 AM
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Hammoud: Muslims must lead by example for peaceful world
A wave of recent attacks by white supremacists has unleashed another barrage of terror upon the Muslim community. Still recovering from the Quebec City mosque massacre, Muslims found themselves mourning the tragic loss of life at the Al-Noor mosque in Christchurch where on March 15, a gunman killed 50 and injured another 50.
Two weeks later, we still know little about the perpetrator or his motives, other than that he had posted online a white supremacist manifesto, live-streamed his attack and engraved his semi-automatic weapon with the name of the perpetrator of the Quebec City attack among other white supremicists.
Perhaps he wanted to create dissension, but what transpired globally after the attack demonstrated a growing intolerance toward radical and violent ideologies. People took to the streets and made human chains around mosques, standing in solidarity with Muslim neighbours, sending a loud, reverberating message to haters everywhere that acts of aggression have no place in our communities.
Home to the first mosque in Ontario and the second in Canada, London always has opened its arms to a diverse and growing community of 30,000 Muslims, and with many international students and visibly identifiable Muslim newcomers from Syria calling it home, there are now half a dozen mosques throughout the city.
Not surprising, days after the attack, Londoners, including elected officials and religious leaders, gathered at a northern entrance of Victoria Park, sadly dubbed "the Corner of Peace," where other vigils also have taken place. They came to offer condolences, words of solace and encouragement to their fellow Muslims, to repeat once again that Londoners stand united against all heinous acts of terror.
Still, in the aftermath of this cowardly attack, Muslims feel vulnerable and afraid to congregate in their mosques, walk in the streets, or live a normal and ordinary life. They don't feel safe anymore.
Today, we stand at dangerous crossroads. Ignorance, rising racial tensions and religious intolerance have turned curiosity into fear. Hate crimes and instances of bullying are not only on the rise, they usually go unreported. A violent vernacular of hate also is stirring feelings of resentment, isolation and eventually more hatred. Sadly, it's evolved well beyond sticks and stones: words of hate breed actions of terror.
While we can't legislate goodness and acceptance in people's hearts, we have the power to change our mindsets and to reform a culture of hatred and terror.
As Muslims, we can start by gaining a greater awareness and understanding of our rights and responsibilities in Canada, learning to advocate on our own behalf as well as on the behalf of our community through our institutions, service providers and the workplace.
We can reject and cast off the false information about Muslims and Islam and reframe a violent vernacular that paints all Muslims as terrorists, a word usually reserved to describe a group who have hijacked Islam to endorse their own perverted interpretation of the faith, a word seldom applied to right-wing nationalists and white supremacists.
Together, with the wider community, we can push for political reform to policies that promote Islamophobia and wider racial injustice. We can strive to better define what constitutes hate speech versus freedom of speech. And we can hold our leaders and elected officials accountable to advocate for anti-racism education and investment in programs that promote diversity.
Finally, as Muslims, we can reclaim our lost identity by speaking up and being heard. We can build bridges, not walls, by extending a hand to our non-Muslim neighbours to say that we are more alike than we are different and although we may come from many different backgrounds, we form one nation, humankind.
Mohamed Hammoud is a London resident and community activist.


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Community News for the Week of November 19th
In this episode, the news that are covered are1. The tentative agreement between childcare workers under CUPE Local 3625 and Something Special Children's Centre after a two week long lock-out2. Laying off of 40 workers from Weston Food in Kingston3. Cuts to education by Doug Ford, the strike votes across the education sector unions and an interview with Andrea Loken, the Chair of the Bargaining Union in OSSTF 67.4. "Density by Design" planning document by the City of Kingston5. A development project by Jay Patry Enterprises on the site of the former Davis Tannery6. Town Hall on housing by MPP Ian Arthur and Katarokwi Union of Tenants' reflection on that7. Issue of homelessness in Kingston, a brief summary of the report by the City of Kingston published last year8. An interview with Bev Woodcock, the director of Special Meals, a hot meal program run by St. Andrew's Church.9. Queen's University's proposed residence build on Albert street and concerns10. 2 new electric buses purchased by the City of Kingston and their contribution to reduction of greenhouse gases11. Advice to keep pets warm in winter by Shevaun Hoyle, the inspector for Kingston Humane Society
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The Place – Small private garden in Karen, Nairobi, Kenya.
The Brief – To design a simple white flower cutting garden around the small cottage.
The Design – This garden is one hundred percent a plants design, we put in simple gravel paths to open up some circulation routes and to frame new


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How to choose a referee
If possible, they should be someone who knows you academically and can talk about your work ethic, interaction with other students and your suitability for higher education or a future career.
- In a current or recent school or college, ask your tutor, teacher, principal or head teacher.
- If you left education years ago, ask an employer, volunteering supervisor or trainer.
- Don't ask family, friends, partners or ex-partners though – if you do your application may be cancelled.
Only one reference is required on the UCAS Undergraduate application. If you want your uni or college to see more than one, you'll need to contact them and ask if an additional reference can be sent directly.
Applying through UCAS Conservatoires? The reference process is different – find out how to get references for UCAS Conservatoires.
Applying through a school, college or centre?
Applying as an individual?
Applying for Teacher Education in Scotland?
Have you been asked to be a referee?
Please see our advice for how to provide a reference and what to include.
How the reference should be written
Here's an overview for you, so that you'll know what to discuss with your referee.
- It must be a full reference, written by your referee online in the reference section of your application, in English (or Welsh if you're applying for courses in Wales).
- It might include your career goals and work experience – so talk these through with them first – as well as how well you've done in specific modules or course components.
- If your referee is one of your teachers, they can add your predicted grades for any pending qualifications you're studying now.
- With your permission, it could include circumstances that have/could affect your academic work, such as a personal problem, illness or any individual needs you have.
Don't want to include a reference?
Applying from prison?


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Duck Cottage Books- Used, Out of Print and Scarce Books
Welcome to Duck Cottage Books. We sell (and buy) used books. You can browse all our books, below, and use the filter in the right sidebar to see books by subject. For a quick search of our stock (searches the book title and description for the word or words you enter) use the search box:
- To search using author, date, publisher etc. go to our Search page.
- To browse books according to subject, go to the Books by Subject page .
- For contact details see our Contact Details page
Showing 1–10 of 2024 results
Practical Notes on Belting and Belt-Driving£30.00 Add to basket
James Stansfeld A Victorian Champion of Sex Equality£20.00 Add to basket
Duty and Delight: Routley Remembered : A Memorial Tribute to Erik Routley, 1917-1982£25.00 Add to basket
The Blackwell Dictionary of Historians£20.00 Add to basket
History of the United Nations War Crimes Commission and The Development Of The Laws Of War£30.00 Add to basket
The Story of Willesden£30.00 Add to basket
Aftermath: a geographical study of the peace terms£12.00 Add to basket
Onward Christian Soldier A Life of Sabine Baring-Gould, Parson, Squire, Novelist, Antiquary 1834-1924£12.50 Add to basket
Prisoner of War£20.00 Add to basket
Crossbow and Overcast£10.00 Add to basket
Showing 1–10 of 2024 results


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Audit Senior
Our client, a prestigious well established Accountancy Practice based in the heart of Central London. An opening has become available for an Audit Senior to join their growing team. The main purpose of this position will be to support the firm's work to ensure that all designated clients receive a first class service as well as developing relationships with existing and potential clients. Duties and Responsibilities:
•Responsible for managing a portfolio of clients
•Completing financial reporting assignments and working directly with and under the tutelage of experienced professional partners
•Leading, planning and completing Statutory Audits from inception to completion
•Preparing financial statements for sole traders, partnerships and limited companies
•Planning and completing a diverse range of assignments
Candidate profile:
•ACCA or ACA qualified or qualified by experience
•Minimum of 4 years experience in audit
Our client is offering an excellent salary package and benefits in a rewarding work environment. Contact us for further information on this position.


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Senior Estimator – Groundworks
- Salary: £50K - £70K
- Location: Ipswich, Suffolk,
- Start date:
- Job Sector:
- Job Type: Permanent
- Job Ref: J3582
Job Spec:
Estimator – Civil Engineering / Groundworks
Ipswich, Suffolk
£55,000 to £65,000 per annum plus car / allowance and benefits package
Our Client is an established Civil Engineering contractor operating successfully in the East Anglia Region. They are renowned for delivering quality on time and to budget and continue winning major projects throughout the region. The company work on residential and commercial groundwork projects. With a strong order book for 2019, they are looking to build on this successful platform with an additional estimator to join their team in preparation for further expansion.
As Senior Estimator you will work closely with other members of the team, including Directors and Project Delivery Teams in the preparation of tenders. The company see this position as a strategic role within the company and can offer a strong career path into senior management for the right person.
• Implement the strategy and systems of the Company, ensuring tenders are undertaken and prepared to the high standards that the Company reputation demands.
• Provide a professional approach to all stages of the tender process, including estimating, tender submission and bid negotiation.
• Ensure that the tender accurately reflects the true cost of labour, plant, materials and sub-contracts, for both direct and in-direct costs.
• Present each tender for Review to Senior Management in sufficient detail to demonstrate confidence in the bid and gain approval to submit the bid.
• Attend site visits and tender meetings to represent the Company and liaise with clients and/or their Representatives throughout the process.
• Previous experience of estimating groundworks packages (£0.25m – £5m contracts).
• Engineering or quantity surveying experience gained within a contracting environment.
• Knowledge of standard methods of measurement.
• Knowledge of standard conditions of contract.
• Strong interpersonal skills.
• Strong IT skills covering Microsoft Word and Excel, and use of Estimating Software is required.
Please contact Ian Farr on 01473 217 399 for a confidential discussion on this and other estimating roles.


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Billionaire business magnate Richard Branson is extremely successful. And at least part of that success can be attributed to the strong teams he's built.
Jas Singh, a recruiting director at Iopa Solutions, says in a LinkedIn post that when building those teams, Branson cares more about a candidate's personality than their skillset.
"Nearly every person mentions how [Virgin] spends extra time and effort getting to know candidates compared to other companies," Singh says in the LinkedIn post, which takes a close look at Branson's success.
While skills can be learned, it's important for Branson to find people who will fit in with Virgin's entrepreneurial culture.
Branson keeps his teams small, spinning off any division that gets too large into it's own company, Singh says. "Smaller teams promote a flatter and more flexible management structure," he explains.
Virgin also encourages employees to pursue their own ideas within the company. "This gives ambitious new hires the belief that they can make a genuine impact to the overall organisation," Singh says.
In a company that's constantly evolving and changing, it makes sense that a candidate's personality would make the biggest impact on the hiring process. Finding employees who are adaptable and innovative will contribute far more than those that simply have a finite set of skills. "Core to [Virgin's] success are the right people," Singh says.
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Looking for something to do in November? We've rounded up a few ideas and events that might just do the trick!
1. Celebrate Malaysia At Riuh In The City
Riuh In The City is back with the Syok-Syok market, a creative and vibrant 2-day event featurine everything Malaysian from flying roti canai to wau making, nasi lemak gelato and more! Besides the aforementioned food and workshops, vendors to look forward to include Apom, Summorie, Kanoe, So.Lek, as well as a variety of performers and an art installation by our very own SuperFEMALE Ruby Subramaniam! Check out the full lineup here.
When: 2 – 3 November 2019
Where: Sentul Depot, Sentul West, 51100 Kuala Lumpur, Federal Territory of Kuala Lumpur
Price: Free admission
Contact: event page


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Root-Server und vServer
Wissen, was der Root-Server und vServer macht mehr »
Ebook Gender Trouble Makers: Education And Empowerment In Nepal (New Approaches In Sociology)by Connie 5
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