The Heavy-Metal Resistance Determinant of Newly Isolated Bacterium from a Nickel-Contaminated Soil in Southwest Slovakia.

A bacterial isolate MR-CH-I2 [KC809939] isolated from soil contaminated mainly by high nickel concentrations in southwest Slovakia was previously found carrying nccA-like heavy-metal resistance determinant, marked as MR-CH-I2-HMR [KF218096]. According to phylogenetic analysis of short (696 bp) 16S rDNA (16S rRNA) sequences this bacterium was tentatively assigned to Uncultured beta proteobacterium clone GC0AA7ZA05PP1 [JQ913301]. nccA-like gene product was on the same base of its partial (581 bp) sequences tentatively assigned to CzcA family heavy metal efflux pump [YP_001899332] from Ralstonia picketii 12J with 99% similarity. In this study the bacterium MR-CH-I2 and its heavy-metal resistance determinant were more precisely identified. This bacterial isolate was on the base of phylogenetic analysis of almost the whole (1,500 bp) 16S rDNA (16S rRNA) sequence, MR-CH-I2 [MF102046], and sequence for gyrB gene and its product respectively, MR-CH-I2-gyrB [MF134666], assigned to R. picketii 12J [CP001068] with 99 and 100% similarities, respectively. In addition, the whole nccA-like heavy-metal resistance gene sequence (3,192 bp), marked as MR-CH-I2-nccA [KR476581], was obtained and on the base of phylogenetic analysis its assignment was confirmed to MULTISPECIES: cation efflux system protein CzcA [WP_004635342] from Burkholderiaceae with 98% similarity. Furthermore, although the bacterium carried one high molecular plasmid of about 50 kb in size, nccA-like gene was not located on this plasmid. Finally, the results from RT-PCR analysis showed that MR-CH-I2-nccA gene was significantly induced only by the addition of nickel.

Heterotrophic microflora of highly alkaline (pH > 13) brown mud disposal site drainage water near Ziar nad Hronom (Banska Bystrica region, Slovakia).

Brown mud is a waste by-product of alumina production by Bayer process. Due to extensive sodium hydroxide use in the process, brown mud disposal site near Ziar nad Hronom (Banska Bystrica region, Slovakia) and drainage water are ones of the greatest environmental burdens in Slovakia. Drainage water from this landfills has pH value higher than 13, and it contains many heavy metals and elevated salt content. In our experiments, relatively numerous bacterial population was detected in the drainage water with frequency of about 80 cfu/ml using cultivation approach. The alkalitolerant heterotrophic isolates were identified by combination of MALDI-TOF and 16S rDNA analysis. Drainage water population was dominated by Actinobacteria (Microbacterium spp. and Micrococcus spp.) followed by low G + C-content gram-positive bacteria (Bacillus spp.). Two isolates belonged to gram-negative bacteria only, identified as Brevundimonas spp. Phylogenetic and biochemical analyses indicate that nearly half of the bacteria isolated are probably representatives of a new species. Brown mud disposal site is proposed as a source of new bacterial taxa possibly used in bioremediation processes.