Alkaline Technosol contaminated by former mining activity and its culturable autochthonous microbiota.

Technosols or technogenic substrates contaminated by potentially toxic elements as a result of iron mining causes not only contamination of the surrounding ecosystem but may also lead to changes of the extent, abundance, structure and activity of soil microbial community. Microbial biomass were significantly inhibited mainly by exceeding limits of potentially toxic metals as arsenic (in the range of 343-511 mg/kg), copper (in the range of 7980-9227 mg/kg), manganese (in the range of 2417-2670 mg/kg), alkaline and strong alkaline pH conditions and minimal contents of organic nutrients. All of the 14 bacterial isolates, belonged to 4 bacterial phyla, Actinobacteria, Firmicutes; ß- and γ-Proteobacteria. Thirteen genera and 20 species of microscopic filamentous fungi were recovered. The most frequently found species belonged to genera Aspergillus (A. clavatus, A. niger, A. flavus, A. versicolor, Aspergillus sp.) with the dominating A. niger in all samples, and Penicillium (P. canescens, P. chrysogenum, P. spinulosum, Penicillium sp.). Fungal plant pathogens occurred in all surface samples. These included Bjerkandera adustata, Bionectria ochloleuca with anamorph state Clonostachys pseudochloleuca, Lewia infectoria, Phoma macrostoma and Rhizoctonia sp.

Wooden art objects and the museum environment: identification and biodegradative characteristics of isolated microflora.

AIMS: The identification of culturable microbial communities on wooden art objects and from indoor air, and the analysis of their biodegradative properties. METHODS AND RESULTS: Common and newly-developed agar media were used for the isolation of fungal and bacterial microflora. The identification was carried out by traditional methods and by the sequencing of 16S or 18S rDNA PCR products. Different plate assays were employed to screen the lignolytic and cellulolytic activities of the isolated microflora. Interesting bacteria were isolated from art objects even though the fungi were the principal contaminants of art works. Various fungal and bacterial species exhibited their lignolytic and cellulolytic activity by the decolorization of Remazol Brilliant Blue R, Phenol Red, Azure B and Ostazin Brilliant Red H-3B. CONCLUSIONS: The microbial communities on wooden art objects exposed in an indoor environment were identified. The study showed the biodegradative power of many microorganisms, and new data were added to this field barely investigated. SIGNIFICANCE AND IMPACT OF THE STUDY: By the development of new culture media and the evaluation of different biodegradative plate assays, a strategy for the analysis of microflora in wooden art objects was established. Several aspects of the study could be also exploited for biotechnology applications.

Regulation of yodA encoding a novel cadmium-induced protein in Escherichia coli.

Bacterial accommodation to moderate concentrations of cadmium is accompanied by transient activation of general stress proteins as well as a sustained induction of other proteins of hitherto unknown functions. One of the latter proteins was previously identified as the product of the Escherichia coli yodA ORF. The yodA ORF encodes 216 aa residues (the YodA protein) and the increased synthesis of YodA during cadmium stress was found probably to be a result of transcriptional activation from one single promoter upstream of the structural yodA gene. Analysis of a transcriptional gene fusion, P(yodA)-lacZ, demonstrated that basal expression of yodA is low during exponential growth and expression is increased greater than 50-fold by addition of cadmium to growing cells. However, challenging cells with additional metals such as zinc, copper, cobalt and nickel did not increase the level of yodA expression. In addition, hydrogen peroxide also increased yodA expression whereas the superoxide-generating agent paraquat failed to do so. Surprisingly, cadmium-induced transcription of yodA is dependent on soxS and fur, but independent of oxyR. Moreover, a double relA spoT mutation abolished induction of yodA during cadmium exposure but ppGpp is not sufficient to induce yodA since expression of the gene is not elevated during stationary phase. After 45 min of cadmium exposure the YodA protein was primarily detected in the cytoplasmic fraction but was later (150 min) found in both the cytoplasmic and periplasmic compartments.

Protein expression in the stressed Vibrio strains.

In a conjunction process using Escherichia coli SM10 (pLOF) KmR APR as donor and Vibrio S141 SmR as recipient, several mutants were constructed: Vibrio PH 101, V. PH 106, and V. PH 109 with lowered ability to synthesize poly-beta-hydroxybutyrate. The survival and metabolic activities of parent and mutant strains were estimated when they were subjected to stress conditions (starvation of carbon and energy sources and/or cadmium treatment). Using two-dimensional electrophoresis, the synthesis of stress proteins was demonstrated. Vibrio cultures consecutively exposed to CdCl2 and then to starvation or vice versa responded similarly metabolically. These results show increased proteosynthetic activity of the stressed Vibrio cells, indicating that the primary cadmium treatment induced the expression and synthesis of the protective proteins, enabling the cells to cope with the secondary stress.