1,721,098 research outputs found
A Helping Hand during -Testing Times.
No full textSiouxsie Wiles reviews by cartoonist Grady Klein and statistician Alan Dabney
How researchers can prepare to make the switch to open science?, figshare fest New Zealand 2018
No full textPresented at figshare fest New Zealand 2018, Wednesday 24th October, hosted at the University of Auckland.Associate Professor Siouxsie Wiles is the Head of the Bioluminescent Superbugs Lab at
the University of Auckland.Event was sponsored by figshare.com</div
Going viral: A science communication collaboration in the era of COVID-19 and social media
No full textOn 9 March 2020, 2 days before the World Health Organization declared COVID-19 a global pandemic, two of the authors (microbiologist and infectious diseases expert Associate Professor Siouxsie Wiles and cartoonist Toby Morris) released their first output together: an animated GIF (Graphics Interchange Format) known as “Flatten the Curve”. The graphic went viral on Twitter with over 10 million impressions in 3 days. Flatten the Curve was the first of more than 70 graphics produced by our collaboration, all designed as accessible visual communication about COVID-19. The graphics, all released under a Creative Commons CC-BY-SA-4.0 license, have been translated into multiple languages, used by communities, politicians, and public health officials around the world, and the collaborators have won multiple awards for their work
Going viral: A science communication collaboration in the era of COVID-19 and social media
No full textOn 9 March 2020, 2 days before the World Health Organization declared COVID-19 a global pandemic, two of the authors (microbiologist and infectious diseases expert Associate Professor Siouxsie Wiles and cartoonist Toby Morris) released their first output together: an animated GIF (Graphics Interchange Format) known as “Flatten the Curve”. The graphic went viral on Twitter with over 10 million impressions in 3 days. Flatten the Curve was the first of more than 70 graphics produced by our collaboration, all designed as accessible visual communication about COVID-19. The graphics, all released under a Creative Commons CC-BY-SA-4.0 license, have been translated into multiple languages, used by communities, politicians, and public health officials around the world, and the collaborators have won multiple awards for their work
Surgical procedure for total knee arthroplasty in mice
No full textDescription of the surgical procedure for total knee arthroplasty used to study impact of intraosseous regional administration of antibiotics
In vivo experimental evolution of ICC180, a bioluminescent derivative of the mouse enteropathogen Citrobacter rodentium - part 2
No full textIn these experiments, we set up 10 transmission chains in which C. rodentium transmitted naturally from mouse to mouse. Experiments were performed as described in the collection provided on Protocols.io (1). In addition, nalidixic acid was added as a treatment for five of the transmission chains. To do this, mice were given nalidixic acid in their drinking water at a final concentration of 10 ug/mL. This water was changed every 2-3 days, adding fresh nalidixic acid each time.This dataset contains the processed data for each animal (1-22) in each transmission chain (N1-5, W1-5) - median weight over the experiment (g), weight range (g), change in weight since the start of the infection (%), weight change at the peak of infection (%), bacterial counts (as colony forming units [CFU] per gram of stool) prior to co-mingling, and calculated area under curve values of bacterial shedding (as cfu per gram of stool.time).1. Read, Patel, & Wiles (2022). Initiating and monitoring natural infection of mice by bioluminescent Citrobacter rodentium. protocols.io. https://www.protocols.io/view/initiating-and-monitoring-natural-infection-of-mic-81wgb6qnnlpk/v1</p
Antimicrobial activity of the endophytic fungus Neofusicoccum australe
No full textTo perform zone of inhibition (ZOI) testing, potato dextrose agar (PDA) plates
were inoculated with a lawn of either antibiotic-sensitive Escherichia coli
(ATCC 25922) or resistant clinical isolates (CTX-M-9, CTX-M-14, CTX-M-15,
NDM-1) or antibiotic-resistant Klebsiella pneumoniae (ATCC 700603).
Similarly, Mueller-Hinton agar plates were inoculated with a lawn of either
antibiotic-sensitive Staphylococcus aureus (ATCC 29213) or
antibiotic-resistant S. aureus (ATCC 33593). Neofusicoccum australe
was grown on PDA plates and fungal plugs removed using a 6 mm punch biopsy tool
(Catalogue number: SH241, Amtech Medical, New Zealand). Fungal plugs were
placed onto the bacterial lawns, alongside PDA plugs containing no fungus. Plates
were incubated inverted at 37°C for 24 h before measuring any zones of
inhibition (in mm) produced
Activity of the pure compounds Asperteretal B, Aspulvinone E, Aspulvinone G, Butyrolactone I, Butyrolactone II, Flavipesolide C, Terretonin, and Terretonin A isolated from the fungus Aspergillus terreus ICMP 477 against Mycobacterium abscessus and M. marinum.
No full text
The fungus Aspergillus terreus ICMP 477 was isolated in September 1961 in Auckland, Aotearoa New Zealand, from sheep’s wool incubated at 30 °C. Forty Potato Dextrose Agar plates were inoculated with ICMP 477 and incubated at room temperature for 3 weeks. Fully grown fungal plates were freeze-dried (26.57 g, dry weight) and extracted with MeOH (2 × 500 mL) for 4 h followed CH2Cl2 (2 × 500 mL) overnight. Combined organic extracts were concentrated under reduced pressure to afford an orange oil (2.45 g). The crude product was subjected to C8 reversed-phase column chromatography eluting with a gradient of H2O/MeOH to give five fractions. The pure compounds were obtained after further fractionation by Sephadex LH-20 and silica gel column chromatography.
Antimicrobial evaluation of the pure compounds was assessed against Mycobacterium abscessus and M. marinum. Because of the slow growth of many mycobacterial species, we routinely use luciferase-tagged strains for our assays. M. abscessus BSG301 and M. marinum BSG101 (1) are stable bioluminescent derivatives transformed with the integrating plasmid pMV306G13ABCDE (2). As bacteria only produce light when alive, bioluminescence is an excellent non-destructive real-time reporter to assay for anti-mycobacterial activity in microtitre plate formats using a luminometer (1,3,4) or in vivo using sensitive imaging equipment (5).
Mycobacterial cultures were grown with shaking (200 rpm) in Middlebrook 7H9 broth (Fort Richard, Auckland) supplemented with 10% Middlebrook ADC enrichment media (Fort Richard), 0.4% glycerol (Sigma-Aldrich) and 0.05% tyloxapol (Sigma-Aldrich). M. abscessus was grown at 37 °C and M. marinum at 28 °C. Cultures were grown until they reached stationary phase (approximately 3-5 days for M. abscessus BSG301 and 7-10 days for M. marinum BSG101) and then diluted in Mueller Hinton broth II (MHB) (Fort Richard) supplemented with 10% Middlebrook ADC enrichment media and 0.05% tyloxapol to give an optical density at 600 nm (OD600) of 0.001 which is the equivalent of ~106 bacteria per mL. Pure compounds were dissolved in DMSO and added in triplicate to the wells of a black 96-well plate (Nunc, Thermo Scientific) at a concentration of 128 μg/mL. Then, 50 μL of diluted bacterial culture was added giving final compound concentrations of 64 μg/mL and a cell density of ~5 × 105 CFU/mL. Rifampicin (Sigma-Aldrich) was used as positive control at 1000 μg/mL for M. abscessus and 10 μg/mL for M. marinum. Between measurements, plates were covered, placed in a plastic box lined with damp paper towels and incubated with shaking at 100 rpm at 37 °C for M. abscessus and 28 °C for M. marinum. Bacterial luminescence (as relative light units (RLU) was measured at regular intervals using a Victor X-3 luminescence plate reader (PerkinElmer) with an integration time of 1 s. More detailed protocols are available at protocols.io (6, 7).
Data is provided as Area Under Curve (AUC) values of luminescence readings for extracts (column = auc) and controls (column = median_ctrl_auc) and corresponding log reduction in AUC comparing extracts and controls (column = log_reduction_auc). Experiments were performed with three technical replicates of one to two biological replicate of each testing bacterium (column = organism [MA, M. abscessus; MM, M. marinum) depending on the quantity of pure compound available.
References:
Dalton JP, Uy B, Okuda K, Hall CJ, Denny WA, Crosier PS, Swift S, Wiles S (2017). Screening of anti-mycobacterial compounds in a naturally infected zebrafish embryo model. Journal of Antimicrobial Chemotherapy 72(2):421-427 (doi: 10.1093/jac/dkw421).
Andreu N, Zelmer A, Fletcher T, Elkington PT, Ward TH, Ripoll J, Parish T, Bancroft GJ, Schaible UE, Robertson BD, Wiles S (2010). Optimisation of bioluminescent reporters for use with Mycobacteria. PLOS One. 5(5): e10777 (doi:10.1371/journal.pone.0010777).
Andreu N, Fletcher T, Krishnan N, Wiles S, Robertson BD (2012). Rapid measurement of antituberculosis drug activity in vitro and in macrophages using bioluminescence. Journal of Antimicrobial Chemotherapy. 67(2): 404-14 (doi: 10.1093/jac/dkr472).
Dalton JP, Uy B, Phummarin N, Copp BR, Denny WA, Crosier PS, Swift S, Wiles S (2016). Effect of common and experimental anti-tuberculosis treatments on Mycobacterium tuberculosis growing as biofilms. PeerJ. 4:e2717 (doi: 10.7717/peerj.2717).
Andreu N, Zelmer A, Sampson SL, Ikeh M, Bancroft GJ, Schaible UE, Wiles S, Robertson BD (2013). Rapid in vivo assessment of drug efficacy against Mycobacterium tuberculosis using an improved firefly luciferase. Journal of Antimicrobial Chemotherapy. 68(9):2118-27 (doi: 10.1093/jac/dkt155).
Grey A & Wiles S (2021). Bioluminescence-based Minimum Inhibitory Concentration (MIC) testing of pure compounds isolated from fungi against Mycobacterium marinum. Protocols.io. (doi: dx.doi.org/10.17504/protocols.io.3x7gprn).
Grey A & Wiles S (2021). Bioluminescence-based Minimum Inhibitory Concentration (MIC) testing of pure compounds isolated from fungi against Mycobacterium abscessus. Protocols.io. (doi: dx.doi.org/10.17504/protocols.io.bumcnu2w).
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