Molecular profiling of Jatropha curcas L. collected from different geographical locations of India

The present study surveys the morphological, biochemical and molecular diversity in 30 accessions of Jatropha collected from different states of India by using random amplified polymorphic DNA (RAPD), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and isozyme analysis. The genotyping data were used to understand the relationships among accessions and to identify genetic diversity as a means for genetic improvement of Jatropha. Out of 37 decamer primers used, 18 yielded polymorphic banding pattern. In total, 126 different DNA bands were reproducibly obtained, out of which 81 were polymorphic. SDS-PAGE of seed, leaf and callus resolved into 18, 12 and 7 bands, while no biotype-specific band was found in leaf and callus protein profile. Cluster analysis of both RAPD and SDS-PAGE data produced two major clusters.     

Effects of fumigants on microbial diversity and persistence of E. coli O15:H7 in contrasting soil microcosms

Persistence of E. coli O157 in the environment is a serious public health concern. However, little is known about the persistence of this pathogen after exposure to chemical compounds like fumigants in the environment. In this study, the persistence behavior of pathogenic E. coli O157:H7 was investigated after fumigation with methyl bromide (MeBr; CH₃Br) and methyl iodide (MeI, iodomethane; CH₃I) in soil microcosms under laboratory conditions. Our goal was to assess changes in soil microbial community structure and persistence of E. coli O157:H7 in microcosm soils after fumigation. PCR was used to amplify 16S rRNA genes from total bacterial community composition, and the products were subjected to denaturing gradient gel electrophoresis (DGGE). Microbial diversity as determined by DGGE was significantly higher in clay soil than sandy soil. Real-time PCR and plate counts were used to quantify the survival of E. coli O157:H7 in the two soils after fumigation with MeBr and MeI. The survival of the pathogen was higher in the non fumigated controls than the fumigated treatments when determined using plate counts. These results were confirmed by real time PCR analysis targeting the stx1, stx2, and the eae genes. E. coli O157:H7 survived for about 35 days when determined using the plate count method but continued to be detected at about the detection limit of 10² by real time PCR for more than 86 days. Our results showed that there was a fast inactivation of the pathogen during the first 35 days. After this period, a small proportion of the pathogen continued to survive in the soil microcosms. Subsequent enrichment of soil samples and immunomagnetic separation revealed the continuous presence of viable cells after 86 days of incubation. The data presented contribute to a better understanding of the behavior of E. coli O157:H7 in soil, and showed the need for more investigation of the role of dormant cells in soil that may be a source for recontamination of the environment.     

Sustainable reduction in the flux of microbial compliance parameters from urban and arable land use to coastal bathing waters by a wetland ecosystem produced by a marine flood defence structure

'Natural' treatment systems such as wetlands and reed beds have been proposed as sustainable means of reducing fluxes of faecal indicator organisms (FIOs) to recreational and shellfish harvesting waters. This is because FIO fluxes to coastal waters from both point (effluent) and diffuse (catchment) sources can cause non-compliance with microbiological standards for bathing and shellfish harvesting waters. The Water Framework Directive requires competent authorities in the member states to manage both point and diffuse sources of FIOs in an integrated manner to achieve compliance with 'good' water quality as defined in a series of daughter Directives. This study was undertaken to investigate the relative sources of FIOs to the popular bathing waters around Clacton, UK. In this predominantly arable (mainly cereal cropping) farming area, the principal land use predictor, explaining 76% of the variance in geometric mean presumptive Escherichia coli concentration at sub-catchment outlets during the bathing season, was the proportion of built-up (i.e. urbanised) land in each sub-catchment. This new finding contrasts with earlier studies in livestock farming regions where the proportion of improved grassland has proven to be the strongest predictor of microbial concentration. Also novel in this investigation, a flood defence wall has been built creating a wetland area which discharges every tidal cycle. The wetland produces over 97% reduction in the flux and concentrations of FIOs to the marine recreational waters. Also, FIO concentrations in water draining through the wetland to the sea were similar to concentrations measured in six UK sewage treatment plant effluents subject to secondary (biological) treatment followed by UV disinfection.