Roy Moate

Dietary mannan oligosaccharide supplementation modulates intestinal microbial ecology and improves gut morphology of rainbow trout, Oncorhynchus mykiss (Walbaum).

A study was conducted to investigate the effect of mannan oligosaccharide (MOS) on the gut microbiota and intestinal morphology of rainbow trout under commercial farming conditions. Juvenile (mean initial BW 38.2 +/- 1.7 g) and subadult (111.7 +/- 11.6 g) trout were fed 2 dietary treatments for 111 and 58 d, respectively. The control treatment consisted of a standard commercial diet, and the MOS treatment consisted of the control diet supplemented with 0.2% MOS. Morphology of the anterior and the posterior intestine was examined with light and electron microscopy. Light microscopy demonstrated increased gut absorptive surface area in the subadult MOS group. Additionally, electron microscopy revealed an increase in microvilli length and density in the subadult MOS group compared with the control (P < 0.05). However, no significant improvements were detected in the juvenile group. Culture-based evaluation of the intestinal microbiota showed that MOS significantly reduced (P < 0.05) the viable intestinal bacterial populations (by approximately 2 log scales in all cases). Levels of Aeromonas/Vibrio spp. were significantly decreased (P < 0.05) in the juvenile MOS group (9% of the total microbiota) compared with the juvenile control group (37%). Additionally, analysis of microbial communities was conducted using denaturing gradient gel electrophoresis of PCR-amplified 16S rDNA. The denaturing gradient gel electrophoresis fingerprinting revealed an alteration of bacterial populations; analysis of similarity, similarity percentages, and nonmetric multidimensional scaling analysis showed that MOS reduced species richness and increased similarity of bacterial populations found within the subadult and juvenile groups. The current study shows that MOS modulates intestinal microbial communities, which subsequently improve gut morphology and epithelial brush border.

The effect of material choice on biofilm formation in a model warm water distribution system

Water distribution systems (WDS) are composed of a variety of materials and may harbour potential pathogens within surface-attached microbial biofilms. Biofilm formation on four plumbing materials, viz. copper, stainless steel 316 (SS316), ethylene propylene diene monomer (EPDM) and cross-linked polyethylene (PEX), was investigated using scanning electron microscope (SEM)/confocal microscopy, ATP-/culture-based analysis, and molecular analysis. Material ‘inserts’ were incorporated into a mains water fed, model WDS. All materials supported biofilm growth to various degrees. After 84 days, copper and SS316 showed no significant overall differences in terms of the level of biofilm formation observed, whilst PEX supported a significantly higher level ofbiofilm. EPDM exhibited gross contamination by a complex, multispecies biofilm, at a level significantly higherthan was observed on the other materials, regardless of the analytical method used. PCR-DGGE analysis showed clear differences in the composition of the biofilm community on all materials after 84 days. The primary conclusion of this study has been to identify EPDM as a potentially unsuitable material for use as a major component in WDS.

Comparison of techniques for the removal of particulate material from seaweed tissue

Eight techniques for cleaning particulate material and epibionts from the tissue of Fucus vesiculosus were tested on seaweed collected from a metal-contaminated estuary in the southwest of England. When assessed by scanning electron microscopy, only one was found to have efficiently removed the particulate material on the tissue. This technique consisted of applying a 1:9 ethanol:seawater mixture to the surface of the thallus, and scraping with a PTFE spatula. Analysis of Cu, As, Fe, Mn, and Zn content in the tissues cleaned by this technique showed that, in this case, only Fe concentrations decreased significantly when compared to tissue cleaned by rinsing in seawater only. The study is the first to systematically assess cleaning techniques for marine macroalgae and illustrates the need for standard methods for the sampling and analysis of seaweed tissue

Arrested development in Fucus spiralis (Phaeophyceae) germlings exposed to copper

Exposure of Fucus spiralis germlings to precise copper concentrations (0 to 844 nM Cu2+) in chemically defined medium demonstrated a relationship between ultrastructural changes and growth retardation with increasing copper concentration. Electron-translucent vesicles, present in ova, which normally disappear after fertilization, accumulated in germlings exposed to Cu2+ above 10.6 nM, suggesting that copper may inhibit a metabolic pathway involved in cell wall formation which is initiated by fertilization. No membrane damage was observed during the exposure period. During a post-exposure period in copper-free medium, recovery occurred (rhizoid extension, apical hair formation) in germlings previously exposed to concentrations below 106 nM Cu2+ and electron-translucent vesicles became granular and disappeared. It is proposed that the electron-translucent vesicles contain a cell wall precursor and that copper inhibits its incorporation into the cell wall, preventing growth and development of the zygote.

Enhanced exchange anisotropy in IrMn/CoFeB systems and its correlation with uncompensated interfacial spins

Bottom pinned exchange bias systems of IrMn/CoFe and IrMn/CoFeB on CoFe seed layers were studied. Enhanced exchange anisotropy has been observed for IrMn/CoFeB samples annealed at 350 °C. The ferromagnetic and antiferromagnetic layers of both samples are polycrystalline and textured {110} for the CoFe and CoFeB, and {111} for IrMn. Results demonstrated that the enhanced exchange anisotropy in the IrMn/CoFeB system is closely associated with the increased uncompensated interfacial spins as evidenced by the enhanced Mn x-ray magnetic circular dichroism (XMCD) signal strength. A quantitative correlation between the Mn XMCD signal and the exchange anisotropy constant Jk was observed.

Current-assisted magnetization switching in a mesoscopic NiFe ring with nanoconstrictions of a wire

A mesoscopic NiFe ring with nanoconstrictions of a wire was fabricated by electron beam lithography and lift-off techniques. Magnetic switching and reversal process have been measured by magnetoresistance as a function of the applied current. It is shown that the applied current has an effect on the switching fields and finally affects the reversal process. The decrease or increase in the switching field from the vortex state to the onion state depends on the electron flow with respect to the direction of domain propagation. The spin in the ring switches from an onion state to the opposite onion state in the low applied current via the double switching process. However, the spin in the ring switches directly from an onion state to the opposite due to the spin torque effect when the applied current is higher than the critical current density (of 107A∕cm2).

Intraocular lens opacification after corneal endothelial keratoplasty: Electron microscopy and x-ray element spectroscopy analysis

Purpose To assess a newly recognized long‐term complication of Descemet‐stripping automated endothelial keratoplasty (DSAEK). Setting Plymouth Royal Eye Infirmary and Plymouth Electron Microscope Centre, Plymouth, United Kingdom. Design Retrospective case series. Methods This study evaluated cases of intraocular lens (IOL) opacification that developed after uneventful DSAEK. None of the IOLs was previously known to opacify. In 1 case, the opacified IOL was explanted and analyzed using detailed light microscopy, scanning electron microscopic (SEM) analysis, and element x‐ray spectroscopy. Results In all 5 cases, the IOL was hydrophilic acrylic and the eye developed IOL anterior surface opacification 4 to 12 months after DSAEK. In 1 eye, the opacification was symptomatic; thus, an IOL exchange was performed. Light microscopy and SEM analysis of the explanted IOL confirmed opacification on the anterior surface and subsurface areas. X‐ray element spectroscopy showed the granules were composed of calcium and phosphorous. Conclusions These cases indicate that IOL opacification after DSAEK is a late, although newly recognized, complication of endothelial keratoplasty. Financial Disclosure No author has a financial or proprietary interest in any material or method mentioned.

Effects of hyperoxia on the permeability of 16HBE14o– cell monolayers – the protective role of antioxidant vitamins E and C

The use of hyperoxia for critically ill patients is associated with adverse impacts resulting in lung injury accompanied by inflammation. The aim of this study was to evaluate aspects of mechanisms that contribute to hyperoxia‐induced disruption of the epithelial permeability barrier, and also the protective effects of the antioxidants α‐tocopherol and ascorbate. 16HBE14o– cells were cultured as monolayers at an air–liquid interface for 6 days, after which transepithelial electrical resistance reached 251.2 ± 4.1 Ω.cm2 (mean ± standard error of the mean). They were then exposed for 24 h to normoxia (21% O2, 5% CO2), hyperoxia (95% O2, 5% CO2), hyperoxia with 10−7 m α‐tocopherol, hyperoxia with 10−7 m ascorbate, hyperoxia with 10−6 m ascorbate, and hyperoxia with a combination of α‐tocopherol and ascorbate (10−7 m and 10−6 m, respectively). Significant reductions (P < 0.05) in transepithelial electrical resistance seen after hyperoxia (with or without antioxidants) were associated with reductions in the levels of zona occludens‐1 (ZO‐1) observed by immunohistochemistry, and downregulation of ZO‐1 expression (P < 0.01) as compared with normoxia. In contrast, the expression levels of interleukin (IL)‐8, IL‐6 and tumour necrosis factor‐α (TNF‐α) were increased after hyperoxia (P < 0.01), and marked increases in the levels of these cytokines (ELISA) were seen in the medium (P < 0.001) as compared with normoxia. The antioxidant vitamins E and C had a partial protective effect against the hyperoxia‐induced reduction in ZO‐1 levels and the increase in levels of the proinflammatory cytokines IL‐8, IL‐6, and TNF‐α. In conclusion, hyperoxia‐induced epithelial disruption is associated with tight junction weakening, and induction of a proinflammatory environment.

Design and validation of a novel quantitative method for rapid bacterial enumeration using programmed stage movement scanning electron microscopy.

The adhesion of bacteria to surgical implants is the first stage of implant infection. The method for detecting bound bacteria is an important consideration in the study of bacterial adherence and colonisation. Enumeration of bacteria by direct visualisation techniques is labour intensive and time consuming. We have developed and validated a method for enumerating bacteria on porous material surfaces using programmed stage movement scanning electron microscopy and compared cumulative counts after 1-10 stage movements with absolute bacterial counts. We describe this method with three commercially sourced meshes used for abdominal wall hernia repair and with three different inoculums of Staphylococcus epidermidis. The results demonstrate significant correlation to the absolute count after five cumulative counts for all meshes analysed. The mean time saved by the cumulative counting method was 1h and 9 min per mesh. We conclude that advances in scanning electron microscopy and the advent of precise automated stage control have facilitated rapid data acquisition for bacterial counting purposes and that five cumulative counts at 1000× or 2500× magnification are a valid quantitative method for enumerating S. epidermidis bacteria on porous surfaces (with a pore size of up to 1.3 mm).

Field Observations on the Effect of a Mannan Oligosaccharide on Mortality and Intestinal Integrity of Sole (Solea senegalensis, Kaup) Infected by Photobacterium damselae subsp. piscicida

This study was conducted in order to investigate the effect of a mannan oligosaccharide (MOS) on the intestinal morphology of sole (Solea senegalensis, Kaup) reared under commercial conditions. The dietary inclusion rate for MOS was 0.4% and it was used either alone or in combination with a vaccination regime against bacterial diseases (Pasteurella spp. and Vibrio spp.). One week after the start of the experimental period, a natural outbreak of pasteurellosis, caused by Photobacterium damselae subsp.piscicida,occurred in all the groups of fish.A two-way ANOVA showed that only MOS supplementation reduced fish mortality by ca. 8% (P = 0.050). Additionally, light microscopy examination of the intestine revealed that MOS supplementation significantly increased the mucosal folding by 29% (P = 0.016) in the anterior intestinal region and by 33% (P = 0.002) in the posterior intestinalregion. Scanning electron microscopy demonstrated that both MOS supplementation and vaccination significantly increased microvilli density on the enterocyte surfaces in the anterior intestinal regionby 13% (P = 0.028) and 30% (P = 0.001) respectively. In the posterior intestinalregion neither MOS supplementation nor vaccination significantly affected the microvilli density (P = 0.005).The present study suggests that dietary MOS supplementation protects the intestinal morphology of infected sole and hinders the development of pathogenic infection, possibly by binding with Photobacterium damselae subsp. piscicida, resulting in reduced mortality of infected fish.