High-resolution melt-curve analysis of random-amplified-polymorphic-DNA markers, for the characterisation of pathogenic Leptospira.

A new test for pathogenic Leptospira isolates, based on RAPD-PCR and high-resolution melt (HRM) analysis (which measures the melting temperature of amplicons in real time, using a fluorescent DNA-binding dye), has recently been developed. A characteristic profile of the amplicons can be used to define serovars or detect genotypes. Ten serovars, of leptospires from the species Leptospira interrogans (serovars Australis, Robinsoni, Hardjo, Pomona, Zanoni, Copenhageni and Szwajizak), L. borgpetersenii (serovar Arborea), L. kirschneri (serovar Cynopteri) and L. weilii (serovar Celledoni), were typed against 13 previously published RAPD primers, using a real-time cycler (the Corbett Life Science RotorGene 6000) and the optimised reagents from a commercial kit (Quantace SensiMix). RAPD-HRM at specific temperatures generated defining amplicon melt profiles for each of the tested serovars. These profiles were evaluated as difference-curve graphs generated using the RotorGene software package, with a cut-off of at least 8 'U' (plus or minus). The results demonstrated that RAPD-HRM can be used to measure serovar diversity and establish identity, with a high degree of stability. The characterisation of Leptospira serotypes using a DNA-based methodology is now possible. As an objective and relatively inexpensive and rapid method of serovar identification, at least for cultured isolates, RAPD-HRM assays show convincing potentia.

Collation, analysis and research of thermal benefits of green life in the urban landscapes

This project reviewed international research conducted on the possible role of plants in alleviating high temperatures in our living spaces. The literature review served to identify the work that has already been carried out in the area and to highlight the gaps to be filled by experimental research. A pilot study then investigated the thermal properties of six of the most common landscaping materials. This project clearly shows that plants can play a significant role in modifying the thermal conditions of urban environments. Tall trees can shade nearby buildings and allow for reductions in cooling costs. In addition to basic shading, the dispersal of heat via the plant’s natural transpiration stream has long been recognised as an important component of the urban energy balance. It has been shown that urban temperatures can be up to 7°C higher than nearby rural areas, illustrating the impact of plants on their environment. These benefits argue against the idea of removing plants from landscapes in order to save on water in times of drought. Similarly, the idea of switching to artificial turf is questionable, since artificial turf still requires watering and can reach temperatures that far exceed the safe range for players. While vegetation offers evaporative cooling, non-vegetative, impervious surfaces such as concrete do not, and can therefore cause greater surface and soil temperatures. In addition, the higher temperatures associated with these impervious surfaces can negatively affect the growth of plants in surrounding areas. Permeable surfaces, such as mulches, have better insulating properties and can prevent excessive heating of the soil. However, they can also lead to an increase in reflected longwave radiation, causing the leaves of plants to close their water-conducting pores and reducing the beneficial cooling effects of transpiration. The results show that the energy balance of our surroundings is complicated and that all components of a landscape will have an impact on thermal conditions.