Bacterial aerosols in a municipal landfill environment.

Landfills collecting substantial amounts of municipal waste support multiplication of different bacteria mainly due to organic matter contained in the deposited materials. With time, they may become active emission sources of these microorganisms. Taking into account both occupational and public health and safety, there is an indisputable necessity to monitor the level of air contamination caused by both bacterial cells and their components (e.g., endotoxins). In this study, the concentrations of total viable bacteria (TVB), and Gram-negative bacteria (GNB), as well as their particle size distributions and concentrations of GNB endotoxins were assessed at various locations within the landfill area. The concentrations of TVB and GNB in the air samples changed depending on the season, location (i.e. active sector versus surroundings) and landfill activity level (i.e. exploitation or standstill periods). Higher abundances of endotoxins were found during the standstill period, and they were significantly correlated with organic dust concentrations. The microbial particle size distribution was associated with the landfill operational state, being predominated by fine below 4.7 µm and coarse fractions above 7.0 µm within the active sector during exploitation and standstill periods, respectively. These results supported by a spatial distribution of bacterial aerosol indicate a clear impact of operated landfill on microbiological air quality within the occupied location and nearby areas. Considering health and safety of landfill workers and neighboring residents, who can be exposed to airborne microbial pollutants, repeated bioaerosol monitoring need to be established. It should facilitate both a special planning within the landfill area and undertaking preventive actions in its near and distant surroundings.

Assessment of relationship between fungal aerosol within a municipal dump and epiphytic mycoflora of crop plants.

A field study was performed to assess whether fungal aerosol of a municipal dump may impact on quantitative and qualitative characteristics of epiphytic mycoflora of crop plants cultivated in vicinity of the dump. Sampling sites were located at every side of the dump. Plant samples were collected from field bean, spring wheat and potato. The highest concentration of fungal aerosol was found at the field located south of the dump within the zone of 250 m next to its borders. For this zone, the most numerous and diverse mycoflora was ascertained, and the plants cultivated were the most damaged. The results suggest that the municipal dump was not the source of phytopathogenic fungi; however, different emissions of contaminants from the dump might cause a decline in the intrinsic plant resistance against the pathogens.

GFP-tagged multimetal-tolerant bacteria and their detection in the rhizosphere of white mustard.

The introduction of rhizobacteria that tolerate heavy metals is a promising approach to support plants involved in phytoextraction and phytostabilisation. In this study, soil of a metal-mine wasteland was analyzed for the presence of metal-tolerant bacterial isolates, and the tolerance patterns of the isolated strains for a number of heavy metals and antibiotics were compared. Several of the multimetal-tolerant strains were tagged with a broad host range reporter plasmid (i.e. pPROBE-NT) bearing a green fluorescent protein marker gene (gfp). Overall, the metal-tolerant isolates were predominately Gram-negative bacteria. Most of the strains showed a tolerance to five metals (Zn, Cu, Ni, Pb and Cd), but with differing tolerance patterns. From among the successfully tagged isolates, we used the transconjugant Pseudomonas putida G25 (pPROBE-NT) to inoculate white mustard seedlings. Despite a significant decrease in transconjugant abundance in the rhizosphere, the gfp-tagged cells survived on the root surfaces at a level previously reported for root colonisers.

Microbial community in the rhizosphere of young maize seedlings is susceptible to the impact of introduced pseudomonads as indicated by FAME analysis.

Two species of Pseudomonas (i.e. P. chlororaphis or P. putida) derived from a maize rhizosphere were studied for their impact on the structure of the microbial community in the rhizosphere of young maize seedlings after inoculation. The culturable bacteria and total microbial communities were analyzed based on profiles of whole-cell fatty acid methyl esters (MIDI-FAME). The introduction of Pseudomonas species resulted in the shift from the Gram-positive dominated culturable community in the rhizosphere of uninoculated maize to more Gram-negative populations in the rhizospheres of the inoculated plants. For the total rhizosphere communities, 43, 47 and 42 FAMEs were detected in the uninoculated maize and the samples inoculated with P. chlororaphis or P. putida, respectively. In contrast to the culturable communities, low concentrations of marker FAMEs for Gram-positives (i15:0, a15:0, i16:0) were found in the profiles of the total rhizosphere communities. The maize inoculations resulted in an enrichment of some Gram-negative isolates; however, Gram-positive bacteria, Cytophaga/Flavobacterium and saprophytic fungi were found in the uninoculated rhizosphere.

Mycorrhizal fungi and ectomycorrhiza associated bacteria isolated from an industrial desert soil protect pine seedlings against Cd(II) impact.

Effects of mycorrhization with Amanita rubescens or Hebeloma sinapizans and dual inoculation with the fungi and ectomycorrhiza associated bacteria (EMAB) Pseudomonas putida or Bacillus cereus on seedling growth and accumulation of Cd(II) in Pinus sylvestris were studied. Both fungal and bacterial species were isolated from roots of pines growing in an industrial area polluted with high concentrations of heavy metals. During mycorrhization, A. rubescens colonized higher number of pine seedlings than H. sinapizans, especially when EMAB were co-inoculated. In addition, the seedling biometric characteristics (i.e. root and shoot lengths and biomass) were stimulated by treatment with the fungal species alone and dual inoculation with the fungi and EMAB. Amanita rubescens was more efficient in this stimulation than H. sinapizans. The increased growth of pine seedlings was especially seen for co-inoculation with P. putida. Furthermore, elevated accumulation of Cd(II), ranging from 56 microg g(-1) to 72 microg g(-1) dry weight, in underground parts of the inoculated seedlings was found. The seedlings treated with A. rubescens accumulated higher concentrations of the metal than those inoculated with H. sinapizans. Additional treatment of pine seedlings with P. putida resulted in the higher accumulation of Cd(II) in the roots as compared with those inoculated with B. cereus. The results suggest that the growth of pine seedlings in Cd(II)-polluted soil may depend on fungal species forming ectomycorrhizae, species-specific co-inoculation with EMAB and specificity of fungal-EMAB interactions.

A polyphasic approach for studying the interaction between Ralstonia solanacearum and potential control agents in the tomato phytosphere.

Ralstonia solanacearum biovar 2, the causative agent of brown rot in potato, has been responsible for large crop losses in Northwest Europe during the last decade. Knowledge on the ecological behaviour of R. solanacearum and its antagonists is required to develop sound procedures for its control and eradication in infested fields.A polyphasic approach was used to study the invasion of plants by a selected R. solanacearum biovar 2 strain, denoted 1609, either or not in combination with the antagonistic strains Pseudomonas corrugata IDV1 and P. fluorescens UA5-40. Thus, this study combined plating (spread and drop plate methods), reporter gene technology (gfp mutants) and serological (imunofluorescence colony staining [IFC]) and molecular techniques (fluorescent in situ hybridization [FISH], PCR with R. solanacearum specific primers and PCR-DGGE on plant DNA extracts). The behaviour of R. solanacearum 1609 and the two control strains was studied in bulk and (tomato) rhizosphere soil and the rhizoplane and stems of tomato plants. The results showed that an interaction between the pathogen and the control strains at the root surface was likely. In particular, R. solanacearum 1609 CFU numbers were significantly reduced on tomato roots treated with P. corrugata IDV1(chr:gfp1) cells as compared to those on untreated roots. Concomitant with the presence of P. corrugata IDV1(chr:gfp1), plant invasion by the pathogen was hampered, but not abolished.PCR-DGGE analyses of the tomato rhizoplane supported the evidence for antagonistic activity against the pathogen; as only weak R. solanacearum 1609 specific bands were detected in profiles derived from mixed systems versus strong bands in profiles from systems containing only the pathogen. Using FISH, a difference in root colonization was demonstrated between the pathogen and one of the two antagonists, i.e. P. corrugata IDV1(chr:gfp1); R. solanacearum strain 1609 was clearly detected in the vascular cylinder of tomato plants, whereas strain IDV1 was absent.R. solanacearum 1609 cells were also detected in stems of plants that had developed in soils treated with this strain, even in cases in which disease symptoms were absent, indicating the occurrence of symptomless infection. In contrast, strain 1609 cells were not found in stems of several plants treated with either one of the two antagonists. The polyphasic analysis is valuable for testing antagonistic strains for approval as biocontrol agents in agricultural practice.