Experiences in fosfomycin susceptibility testing and resistance mechanism determination in Escherichia coli from urinary tract infections in the UK

PURPOSE: With an increase in the numbers of bacterial isolates resistant to first-line antibiotics, there has been a revival in the use of older drugs including fosfomycin with novel mechanisms of action. We aimed to investigate the prevalence and genotypic nature of fosfomycin resistance in Escherichia coli from urinary tract infections (UTIs) using the various methods available in the clinical microbiology laboratory. METHODOLOGY: In total, 1000 culture-positive urine samples were assessed for the presence of E. coli and fosfomycin susceptibility was determined using the MAST Uri system, microbroth dilution, agar dilution and E-test strips.Results/Key findings. Initial investigation using breakpoint susceptibility testing on the MAST Uri system identified 62 of 657 (9.5 %) E. coli isolates as fosfomycin-resistant (MIC≥32 µg ml-1). However, on further testing, a lower rate of eight of the 62 (1.3 %) were robustly confirmed to be resistant using microbroth dilution, agar dilution and E-test strips. These true resistant isolates belonged to diverse E. coli multi-locus sequence types and each had a unique set of chromosomal alterations in genes associated with fosfomycin resistance. Fosfomycin-resistant isolates were not multiply drug resistant and did not carry plasmidic fosfomycin resistance genes. Therefore, the use of fosfomycin may be unlikely to drive selection of a particular clone or movement of transferable resistance genes. CONCLUSION: Fosfomycin remains a viable option for the treatment of E. coli in uncomplicated UTIs; different susceptibility testing platforms can give very different results regarding the prevalence of fosfomycin resistance, with false positives being a potential problem that may unnecessarily limit the use of this agent

Coupling dye-integrated polymeric membranes with smartphone detection to classify bacteria

We report the use of a colorimetric plastic-based device to discriminate four pathogenic bacteria: Klebsiella pneumoniae, Proteus vulgaris, Proteus mirabilis and Escherichia coli. The colour changes of the plastic membranes were analysed with RGB values extracted from dye-integrated polymeric membrane images obtained with a smartphone and used as input data for non-supervised pattern recognition methods. © The Royal Society of Chemistry 2015

Functional genomics to identify the factors contributing to successful persistence and global spread of an antibiotic resistance plasmid

Background: The spread of bacterial plasmids is an increasing global problem contributing to the widespread dissemination of antibiotic resistance genes including β-lactamases. Our understanding of the details of the biological mechanisms by which these natural plasmids are able to persist in bacterial populations and are able to establish themselves in new hosts via conjugative transfer is very poor. We recently identified and sequenced a globally successful plasmid, pCT, conferring β-lactam resistance. Results: Here, we investigated six plasmid encoded factors (tra and pil loci; rci shufflon recombinase, a putative sigma factor, a putative parB partitioning gene and a pndACB toxin-antitoxin system) hypothesised to contribute to the 'evolutionary success' of plasmid pCT. Using a functional genomics approach, the role of these loci was investigated by systematically inactivating each region and examining the impact on plasmid persistence, conjugation and bacterial host biology. While the tra locus was found to be essential for all pCT conjugative transfer, the second conjugation (pil) locus was found to increase conjugation frequencies in liquid media to particular bacterial host recipients (determined in part by the rci shufflon recombinase). Inactivation of the pCT pndACB system and parB did not reduce the stability of this plasmid. Conclusions: Our findings suggest the success of pCT may be due to a combination of factors including plasmid stability within a range of bacterial hosts, a lack of a fitness burden and efficient transfer rates to new bacterial hosts rather than the presence of a particular gene or phenotype transferred to the host. The methodology used in our study could be applied to other 'successful' globally distributed plasmids to discover the role of currently unknown plasmid backbone genes or to investigate other factors which allow these elements to persist and spread

Environmentally friendly analysis of emerging contaminants by pressurized hot water extraction-stir bar sorptive extraction-derivatization and gas chromatography-mass spectrometry

This work describes the development, optimiza- tion, and validation of a new method for the simultaneous determination of a wide range of pharmaceuticals (beta- blockers, lipid regulators ... ) and personal care products (fragrances, UV filters, phthalates ... ) in both aqueous and solid environmental matrices. Target compounds were extracted from sediments using pressurized hot water ex- traction followed by stir bar sorptive extraction. The first stage was performed at 1,500 psi during three static extrac- tion cycles of 5 min each after optimizing the extraction temperature (50 – 150 °C) and addition of organic modifiers (% methanol) to water, the extraction solvent. Next, aqueous extracts and water samples were processed using polydime- thylsiloxane bars. Several parameters were optimized for this technique, including extraction and desorption time, ionic strength, presence of organic modifiers, and pH. Fi- nally, analytes were extracted from the bars by ultrasonic irradiation using a reduced amount of solvent (0.2 mL) prior to derivatization and gas chromatography – mass spectrome- try analysis. The optimized protocol uses minimal amounts of organic solvents (<10 mL/sample) and time ( ≈ 8 h/sam- ple) compared to previous ex isting methodologies. Low standard deviation (usually below 10 %) and limits of de- tection (sub-ppb) vouch for the applicability of the method- ology for the analysis of target compounds at trace levels. Once developed, the method was applied to determin

Determining the distribution of triclosan and methyl triclosan in estuarine settings

We have developed a method for the analysis of two sewage-derived contaminants: triclosan (TCS), an antibacterial agent, and methyl triclosan (MTCS), a TCS metabolite. For solid samples (4 g), extraction and cleanup were integrated into the same step using pressurized liquid extraction (PLE) with in-cell-clean-up (1 g of florisil). The extraction was performed using dichloromethane at 100 °C, 1500 psi and 3 static extraction cycles of 5 min each. For water samples (100 mL), stir bar sorptive extraction–liquid desorption (SBSE–LD) was used. Bars were stirred for 10 h and analytes were later desorbed using acetonitrile. Finally, MTCS and a silylated derivative of TCS were determined by gas chromatography–mass spectrometry (GC–MS). Recovery experiments in water and sediments were performed and the results ranged from 67% to 78%. Limits of detection (LODs) were 5 ng L−1 for TCS and 1 ng L−1 for MTCS, in water samples, and 0.1 ng g−1 for TCS and MTCS in solid samples. The method was applied then to determine the levels of these compounds in the estuary of Guadalete River (SW Spain). TCS and MTCS concentrations up to 9.6 ng g−1 in sediments and 310 ng L−1 in water were measured. Their distribution was strongly influenced by the presence of wastewater sources, treated and untreated, along the sampling area, where maximum concentrations were detected. Highest values were reached in the water column during low tides as the water volume in the estuary becomes lower

An imbalance in progenitor cell populations reflects tumour progression in breast cancer primary culture models

Many factors influence breast cancer progression, including the ability of progenitor cells to sustain or increase net tumour cell numbers. Our aim was to define whether alterations in putative progenitor populations could predict clinicopathological factors of prognostic importance for cancer progression.Journal ArticleResearch Support, Non-U.S. Gov'tinfo:eu-repo/semantics/publishe

Antimicrobials in animal agriculture: Parables and policy

In addition to the scientific, economic, regulatory and other policy factors that impact on antimicrobial decision-making in different jurisdictions around the world, there exist ethical, social and cultural bases for the contemporary use of these products in animal agriculture. Thus, the use of the word ‘parable’ to describe the contemporary moral stories that help to guide ethical antimicrobial use practices and broader policy decisions in animal agriculture is appropriate. Several of these stories reflect difficult decisions that arise from conflicting moral imperatives (i.e. both towards animal welfare and towards human health). Understanding the factors that combine to define the past and present paradigms of antimicrobial usage is crucial to mapping a path forward. There exist barriers, as well as opportunities, for advancing scenarios for reducing antimicrobial usage under a variety of voluntary, regulatory and legal policy frameworks. Any new approaches will ideally be structured to extend the use of present-day antimicrobials into the future, to provide novel alternatives for regulating any newly introduced antimicrobial products so as to maximize their useful life span and to ensure the optimal use of these products in animal agriculture to protect not only the health of animals and the interests of animal health/agriculture stakeholders, but also the human health and the interests of the public at large. A full range of policy approaches, which span the realm from strictly enforced regulations and laws to voluntary guidelines and compliance, should be explored with respect to their risks and benefits in a variety of worldwide settings and in full consideration of a range of stakeholder values

Live to cheat another day: bacterial dormancy facilitates the social exploitation of beta-lactamases

The breakdown of antibiotics by β-lactamases may be cooperative, since resistant cells can detoxify their environment and facilitate the growth of susceptible neighbours. However, previous studies of this phenomenon have used artificial bacterial vectors or engineered bacteria to increase the secretion of β-lactamases from cells. Here, we investigated whether a broad-spectrum β-lactamase gene carried by a naturally occurring plasmid (pCT) is cooperative under a range of conditions. In ordinary batch culture on solid media, there was little or no evidence that resistant bacteria could protect susceptible cells from ampicillin, although resistant colonies could locally detoxify this growth medium. However, when susceptible cells were inoculated at high densities, late-appearing phenotypically susceptible bacteria grew in the vicinity of resistant colonies. We infer that persisters, cells that have survived antibiotics by undergoing a period of dormancy, founded these satellite colonies. The number of persister colonies was positively correlated with the density of resistant colonies and increased as antibiotic concentrations decreased. We argue that detoxification can be cooperative under a limited range of conditions: if the toxins are bacteriostatic rather than bacteridical; or if susceptible cells invade communities after resistant bacteria; or if dormancy allows susceptible cells to avoid bactericides. Resistance and tolerance were previously thought to be independent solutions for surviving antibiotics. Here, we show that these are interacting strategies: the presence of bacteria adopting one solution can have substantial effects on the fitness of their neighbours

Personalized ovarian stimulation for assisted reproductive technology : study design considerations to move from hype to added value for patients

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Analisis Pelaksanaan Audit Internal terhadap Pengendalian Internal Penerimaan & Pengeluaran Kas dalam Rangka Menemukan Fraud - Kasus pada PT XYZ

Penelitian ini bertujuan untuk mengetahui apakah teknik audit internal yang diterapkan oleh PT XYZ di Jakarta sebagai aktivitas pemeriksaan dan penilaian Perusahaan secara menyeluruh yang membantu semua tingkatan manajemen dalam melaksanakan tanggung jawabnya telah berjalan secara memadai pada penerimaan kas dan pengeluaran kas. Dalam penelitian ini metode analisis data yang digunakan adalah pendekatan kualitatif. Data primer didapat dari responden melalui observasi, kuesioner, dan wawancara yang berprofesi sebagai auditor internal. Data sekunder didapat melalui dokumen yang berasal dari Perusahaan. Berdasarkan penelitian yang telah dilakukan pada PT XYZ di Jakarta, dapat disimpulkan bahwa inspeksi sebagai teknik audit internal yang digunakan dalam menemukan fraud pada penerimaan kas dan pengeluaran kas telah berjalan secara memadai. Kata kunci: teknik audit internal, penerimaan kas, pengeluaran kas This research aims to determine whether the internal audit technique applied by XYZ Company in Jakarta as the activity and examination of the company\u27s overall assessment that helps all levels of management in carrying out its responsibilities adequately run well on the cash receipt and cash disbursement. In this research, data analysis method used is the qualitative approach. The primary data obtained from the respondents by doing observation, questionnaires, and interviews who works as internal auditor. And then, the secondary data obtained through documents derived from the company. Based on the research which has been done at the XYZ Company in Jakarta, it could be concluded that inspection as the internal audit technique which used in detecting fraud at cash receipt and cash disbursement has been adequately well. Keywords: Internal audit technique, cash receipt, cash disbursement Daftar Pustaka Agoes, Sukrisno. 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