Visualisation and biovolume quantification in the characterisation of biofilm formation in Mycoplasma fermentans.

The ability of mycoplasmas to persist on surfaces has been widely acknowledged, despite their fastidious nature. However, the organism's capability to form a recognisable biofilm structure has been identified more recently. In the current study Mycoplasma fermentans was found to adhere to the glass surface forming highly differentiated biofilm structures. The volumes of biofilm microcolonies were quantified and observed to be greater at late growth stage than those at early growth stage. The channel diameters within biofilms were measured with Scanning Electron Microscopy images and found to be consistent with the size observed in Confocal Laser Scanning Microscope images. The combination of imaging methods with 3D visualisation provides key findings that aid understanding of the mycoplasma biofilm formation and true biofilm architecture. The observations reported here provide better understanding of the persistence of these minimalist pathogens in nature and clinical settings.

Determination of metabolic activity in planktonic and biofilm cells of Mycoplasma fermentans and Mycoplasma pneumoniae by nuclear magnetic resonance.

Mycoplasmas are fastidious microorganisms, typically characterised by their restricted metabolism and minimalist genome. Although there is reported evidence that some mycoplasmas can develop biofilms little is known about any differences in metabolism in these organisms between the growth states. A systematic metabolomics approach may help clarify differences associated between planktonic and biofilm associated mycoplasmas. In the current study, the metabolomics of two different mycoplasmas of clinical importance (Mycoplasma pneumoniae and Mycoplasma fermentans) were examined using a novel approach involving nuclear magnetic resonance spectroscopy and principle component analysis. Characterisation of metabolic changes was facilitated through the generation of high-density metabolite data and diffusion-ordered spectroscopy that provided the size and structural information of the molecules under examination. This enabled the discrimination between biofilms and planktonic states for the metabolomic profiles of both organisms. This work identified clear biofilm/planktonic differences in metabolite composition for both clinical mycoplasmas and the outcomes serve to establish a baseline understanding of the changes in metabolism observed in these pathogens in their different growth states. This may offer insight into how these organisms are capable of exploiting and persisting in different niches and so facilitate their survival in the clinical setting.

Determination of the antioxidant capacity of culinary herbs subjected to various cooking and storage processes using the ABTS(*+) radical cation assay.

Culinary herbs have the potential to be a significant source of antioxidants in the diet. However, many culinary herbs are cooked or undergo some other form of processing before they are consumed as part of a meal and such factors may affect their significance as a source of dietary antioxidants. Thus, the impact of cooking (simmering, microwaving, stewing, stir frying and grilling) and storage (vinegar maceration, cold maceration and freezing) on the antioxidant capacity of common culinary herbs was investigated. Extracts of cinnamon, cloves, fennel, ginger, lavender, parsley, rose, rosemary, sage and thyme were prepared pre and post cooking or storage and their antioxidant capacities determined using the Trolox equivalent antioxidant capacity assay (TEAC). Simmering, soup making and stewing significantly increased antioxidant capacity, whilst grilling and stir frying decreased it. Both freezing herbs at -20 degrees C and cold maceration had preservative effects on antioxidant capacity. Herbs in cold vinegar macerations for 1 week showed a decrease in antioxidant capacity compared to the control extracts. These results indicate that the potential of culinary herbs to be significant contributors to dietary antioxidant intake is significantly affected by both cooking and storage.