The development of a ligase mediated PCR with potential for the differentiation of serovars within Leptospira interrogans

A ligase mediated polymerase chain reaction (LMPCR) was developed to amplify between the repetitive element, IS1533, of Leptospira and adjacent chromosomally located BglII restriction endonuclease enzyme sites. To do this, complimentary oligonucleotide linkers designed to anneal together with an overhanging BglII end were ligated to BglII digested DNA from 35 leptospiral reference strains and field isolates, This ligated DNA was used as template for PCR with oligonucleotide primers specific for the linker and for the repetitive element IS1533. The resultant amplicon profile hybridised a 102 hp region derived from the terminus of IS1533 thus confirming that amplicons generated by LMPCR contained part of IS1533. The number of fragments generated containing IS1533 was significantly fewer than that generated by RFLP but the LMPCR method has the potential to use far less template DNA and be quicker than standard RFLP. Obvious and reproducible interserovar differences were demonstrated by LMPCR whereas for 20 of 21 L. hardjo-bovis isolates tested no intraserovar differences were observed. Of those serovars known to possess IS1533 homologues and tested here by LMPCR, each produced a unique amplicon profile which hybridised the IS1533 terminus probe. The limited heterogeneity amongst hardjo-bovis isolates is discussed as is the potential contribution of this method to diagnosis, differentiation and the phylogenetics of the Leptospires.

On the development of strong ridge episodes over the eastern North Atlantic

The occurrence of strong and persistent mid-latitude anticyclonic ridges over the Eastern North Atlantic is a major contributor to the occurrence of severe winter droughts over Western Iberia. We analyze the development of strong and persistent ridge episodes within 40–50°N; 40°W–5°E, which are defined as 300 hPa geopotential height anomalies above 50 gpm that persist for at least 10 consecutive days. Results suggest that the generation and maintenance of these episodes, with positive stratospheric geopotential anomalies over the North American continent and the adjacent North Pacific, are associated with an intensified polar jet. Such positive anomalies tend to detach from the main stratospheric anomaly and propagate eastwards and downwards as Rossby tropospheric waves. Furthermore, the Eastern North Atlantic ridge is generated and repeatedly reinforced until the stratospheric anomaly dissipates. Results also show evidence for waves breaking anticyclonically during the episodes, which is dynamically coherent with their persistency and quasi-stationarity.