Para-cresol production by Clostridium difficile affects microbial diversity and membrane integrity of Gram-negative bacteria

Clostridium difficile is a Gram-positive spore-forming anaerobe and a major cause of antibiotic-associated diarrhoea. Disruption of the commensal microbiota, such as through treatment with broad-spectrum antibiotics, is a critical precursor for colonisation by C. difficile and subsequent disease. Furthermore, failure of the gut microbiota to recover colonisation resistance can result in recurrence of infection. An unusual characteristic of C. difficile among gut bacteria is its ability to produce the bacteriostatic compound para-cresol (p-cresol) through fermentation of tyrosine. Here, we demonstrate that the ability of C. difficile to produce p-cresol in vitro provides a competitive advantage over gut bacteria including Escherichia coli, Klebsiella oxytoca and Bacteroides thetaiotaomicron. Metabolic profiling of competitive co-cultures revealed that acetate, alanine, butyrate, isobutyrate, p-cresol and p-hydroxyphenylacetate were the main metabolites responsible for differentiating the parent strain C. difficile (630Δerm) from a defined mutant deficient in p-cresol production. Moreover, we show that the p-cresol mutant displays a fitness defect in a mouse relapse model of C. difficile infection (CDI). Analysis of the microbiome from this mouse model of CDI demonstrates that colonisation by the p-cresol mutant results in a distinctly altered intestinal microbiota, and metabolic profile, with a greater representation of Gammaproteobacteria, including the Pseudomonales and Enterobacteriales. We demonstrate that Gammaproteobacteria are susceptible to exogenous p-cresol in vitro and that there is a clear divide between bacterial Phyla and their susceptibility to p-cresol. In general, Gram-negative species were relatively sensitive to p-cresol, whereas Gram-positive species were more tolerant. This study demonstrates that production of p-cresol by C. difficile has an effect on the viability of intestinal bacteria as well as the major metabolites produced in vitro. These observations are upheld in a mouse model of CDI, in which p-cresol production affects the biodiversity of gut microbiota and faecal metabolite profiles, suggesting that p-cresol production contributes to C. difficile survival and pathogenesis.Peer reviewedFinal Published versio

Short-term genome stability of serial Clostridium difficile ribotype 027 isolates in an experimental gut model and recurrent human disease

Copyright: © 2013 Eyre et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are creditedClostridium difficile whole genome sequencing has the potential to identify related isolates, even among otherwise indistinguishable strains, but interpretation depends on understanding genomic variation within isolates and individuals.Serial isolates from two scenarios were whole genome sequenced. Firstly, 62 isolates from 29 timepoints from three in vitro gut models, inoculated with a NAP1/027 strain. Secondly, 122 isolates from 44 patients (2–8 samples/patient) with mostly recurrent/on-going symptomatic NAP-1/027 C. difficile infection. Reference-based mapping was used to identify single nucleotide variants (SNVs).Across three gut model inductions, two with antibiotic treatment, total 137 days, only two new SNVs became established. Pre-existing minority SNVs became dominant in two models. Several SNVs were detected, only present in the minority of colonies at one/two timepoints. The median (inter-quartile range) [range] time between patients’ first and last samples was 60 (29.5–118.5) [0–561] days. Within-patient C. difficile evolution was 0.45 SNVs/called genome/year (95%CI 0.00–1.28) and within-host diversity was 0.28 SNVs/called genome (0.05–0.53). 26/28 gut model and patient SNVs were non-synonymous, affecting a range of gene targets.The consistency of whole genome sequencing data from gut model C. difficile isolates, and the high stability of genomic sequences in isolates from patients, supports the use of whole genome sequencing in detailed transmission investigations.Peer reviewe

The global energy balance of Titan

The global energy budget of planets and their moons is a critical factor to influence the climate change on these objects. Here we report the first measurement of the global emitted power of Titan. Long-term (2004–2010) observations conducted by the Composite Infrared Spectrometer (CIRS) onboard Cassini reveal that the total emitted power by Titan is (2.84 ± 0.01) × 10^(14) watts. Together with previous measurements of the global absorbed solar power of Titan, the CIRS measurements indicate that the global energy budget of Titan is in equilibrium within measurement error. The uncertainty in the absorbed solar energy places an upper limit on the energy imbalance of 6.0%

Chromophores from photolyzed ammonia reacting with acetylene: Application to Jupiter's Great Red Spot

The high altitude of Jupiter's Great Red Spot (GRS) may enhance the upward flux of gaseous ammonia (NH3) into the high troposphere, where NH3 molecules can be photodissociated and initiate a chain of chemical reactions with downwelling acetylene molecules (C2H2). These reactions, experimentally studied earlier by (Ferris and Ishikawa [1987] Nature 326, 777-778) and (Ferris and Ishikawa [1988] J. Amer. Chem. Soc. 110, 4306-4312), produce chromophores that absorb in the visible and ultraviolet regions. In this work we photolyzed mixtures of NH3 and C2H2 using ultraviolet radiation with a wavelength of 214 nm and measured the spectral transmission of the deposited films in the visible region (400-740 nm). From these transmission data we estimated the imaginary indices of refraction. Assuming that ammonia grains at the top of the GRS clouds are coated with this material, we performed layered sphere and radiative transfer calculations to predict GRS reflection spectra. Comparison of those results with observed and previously unreported Cassini visible spectra and with true-color images of the GRS show that the unknown GRS chromophore is spectrally consistent with the coupled NH3-C2H2 photochemical products produced in our laboratory experiments. Using high-resolution mass spectrometry and infrared spectroscopy we infer that the chromophore-containing residue is composed of aliphatic azine, azo, and diazo compounds. <P /

Vortices in Saturn's Northern Hemisphere (2008-2015) observed by Cassini ISS

We use observations from the Imaging Science Subsystem on Cassini to create maps of Saturn's Northern Hemisphere (NH) from 2008 to 2015, a time period including a seasonal transition (i.e., spring equinox in 2009) and the 2010 giant storm. The processed maps are used to investigate vortices in the NH during the period of 2008–2015. All recorded vortices have diameters (east‐west) smaller than 6000 km except for the largest vortex that developed from the 2010 giant storm. The largest vortex decreased its diameter from ~11,000 km in 2011 to ~5000 km in 2015, and its average diameter is ~6500 km during the period of 2011–2015. The largest vortex lasts at least 4 years, which is much longer than the lifetimes of most vortices (less than 1 year). The largest vortex drifts to north, which can be explained by the beta drift effect. The number of vortices displays varying behaviors in the meridional direction, in which the 2010 giant storm significantly affects the generation and development of vortices in the middle latitudes (25–45°N). In the higher latitudes (45–90°N), the number of vortices also displays strong temporal variations. The solar flux and the internal heat do not directly contribute to the vortex activities, leaving the temporal variations of vortices in the higher latitudes (45–90°N) unexplained

Emitted Power Of Jupiter Based On Cassini CIRS And VIMS Observations

The emitted power of Jupiter and its meridional distribution are determined from observations by the Composite Infrared Spectrometer (CIRS) and Visual and Infrared Spectrometer (VIMS) onboard Cassini during its flyby en route to Saturn in late 2000 and early 2001. Jupiter's global- average emitted power and effective temperature are measured to be 14.10+/-0.03 W/sq m and 125.57+/-0.07 K, respectively. On a global scale, Jupiter's 5-micron thermal emission contributes approx. 0.7+/-0.1 % to the total emitted power at the global scale, but it can reach approx. 1.9+/-0.6% at 15degN. The meridional distribution of emitted power shows a significant asymmetry between the two hemispheres with the emitted power in the northern hemisphere 3.0+/-0.3% larger than that in the southern hemisphere. Such an asymmetry shown in the Cassini epoch (2000-01) is not present during the Voyager epoch (1979). In addition, the global-average emitted power increased approx. 3.8+/-1.0% between the two epochs. The temporal variation of Jupiter's total emitted power is mainly due to the warming of atmospheric layers around the pressure level of 200 mbar. The temporal variation of emitted power was also discovered on Saturn (Li et al., 2010). Therefore, we suggest that the varying emitted power is a common phenomenon on the giant planets

Effect of sugammadex on processed EEG parameters in patients undergoing robot-assisted radical prostatectomy

Background: Sugammadex has been associated with increases in the bispectral index (BIS). We evaluated the effects of sugammadex administration on quantitative electroencephalographic (EEG) and electromyographic (EMG) measures. Methods: We performed a prospective observational study of adult male patients undergoing robot-assisted radical prostatectomy. All patients received a sevoflurane-based general anaesthetic and a continuous infusion of rocuronium, which was reversed with 2 mg kg1 of sugammadex i.v. BIS, EEG, and EMG measures were captured with the BIS Vista™ monitor. Results: Twenty-five patients were included in this study. Compared with baseline, BIS increased at 4e6 min (b coefficient: 3.63; 95% confidence interval [CI]: 2.22e5.04; P<0.001), spectral edge frequency 95 (SEF95) increased at 2e4 min (b coefficient: 0.29; 95% CI: 0.05e0.52; P¼0.016) and 4e6 min (b coefficient: 0.71; 95% CI: 0.47e0.94; P<0.001), and EMG increased at 4e6 min (b coefficient: 1.91; 95% CI: 1.00e2.81; P<0.001) after sugammadex administration. Compared with baseline, increased beta power was observed at 2e4 min (b coefficient: 93; 95% CI: 1e185; P¼0.046) and 4e6 min (b coefficient: 208; 95% CI: 116e300; P<0.001), and decreased delta power was observed at 4e6 min (b coefficient: 526.72; 95% CI: 778 to 276; P<0.001) after sugammadex administration. Neither SEF95 nor frequency band data analysis adjusted for EMG showed substantial differences. None of the patients showed clinical signs of awakening. Conclusions: After neuromuscular block reversal with 2 mg kg1 sugammadex, BIS, SEF95, EMG, and beta power showed small but statistically significant increases over time, while delta power decreased

High flavonoid accompanied with high starch accumulation triggered by nutrient starvation in bioenergy crop duckweed (Landoltia punctata)

Background: As the fastest growing plant, duckweed can thrive on anthropogenic wastewater. The purple-backed duckweed, Landoltia punctata, is rich in starch and flavonoids. However, the molecular biological basis of high flavonoid and low lignin content remains largely unknown, as does the best method to combine nutrients removed from sewage and the utilization value improvement of duckweed biomass. Results: A combined omics study was performed to investigate the biosynthesis of flavonoid and the metabolic flux changes in L. punctata grown in different culture medium. Phenylalanine metabolism related transcripts were identified and carefully analyzed. Expression quantification results showed that most of the flavonoid biosynthetic transcripts were relatively highly expressed, while most lignin-related transcripts were poorly expressed or failed to be detected by iTRAQ based proteomic analyses. This explains why duckweed has a much lower lignin percentage and higher flavonoid content than most other plants. Growing in distilled water, expression of most flavonoid-related transcripts were increased, while most were decreased in uniconazole treated L. punctata (1/6 x Hoagland + 800 mg center dot L-1 uniconazole). When L. punctata was cultivated in full nutrient medium (1/6 x Hoagland), more than half of these transcripts were increased, however others were suppressed. Metabolome results showed that a total of 20 flavonoid compounds were separated by HPLC in L. punctata grown in uniconazole and full nutrient medium. The quantities of all 20 compounds were decreased by uniconazole, while 11 were increased and 6 decreased when grown in full nutrient medium. Nutrient starvation resulted in an obvious purple accumulation on the underside of each frond. Conclusions: The high flavonoid and low lignin content of L. punctata appears to be predominantly caused by the flavonoid-directed metabolic flux. Nutrient starvation is the best option to obtain high starch and flavonoid accumulation simultaneously in a short time for biofuels fermentation and natural products isolation

Ecological interactions between Antarctic krill (Euphausia superba) and baleen whales in the South Sandwich Islands region – Exploring predator-prey biomass ratios

Following the cessation of whaling, the southwest Atlantic humpback whale (Megaptera novaeangliae) population is thought to be close to pre-exploitation size, reversing 20th century changes in abundance. Using a model-based approach applied to concurrently collected data on baleen whale abundance and Antarctic krill (Euphausia superba) biomass in the South Sandwich Islands (SSI) region, we explore ecological interactions between these taxa. Krill biomass and baleen whale density were highest to the north and northeast of the SSI, where the Antarctic Circumpolar Current (ACC) is deflected around the island chain. Humpback whale density was elevated at locations of krill biomass density >150 gm-2. Krill consumption by baleen whales was estimated at 19–29% of the available krill standing stock. We used historic whaling data to confirm the plausibility of these consumption rates and found evidence of rapid weight gain in humpback whales, such that blubber depleted during the breeding season could be restored in a much shorter period than previously assumed. Little is known about krill replenishment rates in the flow of the ACC, or about niche separation between recovering baleen whale populations; both factors may affect species carrying capacities and further monitoring will be required to inform the management of human activities in the region