Aims: The former Mamut Copper Mine, acid mine drainage site represents an anthropogenic altered landscape characterized by its acidic topsoil which is contaminated primarily with copper. Even though the mining operation was ceased at 1999, the bacterial diversity in this area has never been investigated. This study was conducted to ascertain the bacterial diversity of this abandoned copper mine and correlate it to the copper concentration in the soil. Methodology and results: Soil samples were collected from 7 sites near the mine pit and the vicinity. Soil samples were assessed for soil copper elemental concentration using inductively coupled plasma optical emission spectrometry and bacteria were isolated via serial dilution followed by culture on nutrient agar plates. Phylogenetic analysis was done based on the full-length sequences of 16S rRNA gene. Twenty-four phylotypes were obtained from the 7 locations which originated from the phyla Firmicutes, Actinobacteria, Bacteroidetes and Proteobacteria. The results of the study indicated that site 2 (6.030223°; 116.658030°), located in between the mine pit and the mine factory with a copper concentration of 88.96 ppm, possessed the most diverse bacterial community with a Shannon diversity index (H) of 1.68, evenness (EH) of 0.94 and richness (S) of 6. Conclusion, significance and impact of study Current study revealed that there was a positive correlation between the copper concentration and the H index and the richness, but this was not reflected in the evenness. This is the first report of bacterial diversity from the former Mamut Copper Mine site. The data provided a valuable insight for the future monitoring of the bacterial community in this ecologically important niche.
Soil Microbiology

Aims: Bacillus thuringiensis Y1 is a copper-tolerant bacterium that can serve as a model for the elucidation of the mechanism of energy metabolism under simulated copper stress. This study aimed to elucidate the effects of simulated copper stress on the genes associated with the biosynthesis of polyhydroxyalkanoates (PAH) and the metabolism of polyphosphates (PP). Methodology and results : The gene expression study involved the growth of the bacterium in nutrient broth supplemented with two concentrations of copper sulphate (0.4 mM and 0.8 mM), followed by RNA extraction and quantification of four and 11 genes associated with the metabolism of polyphosphate (PP) and polyhydroxyalkanoates (PAHs) respectively, using reverse transcription quantitative polymerase chain reaction (RT-qPCR). Three genes associated with polyphosphate metabolism, which are polyphosphate kinase (ppk), exopolyphosphatase (ppx) and NAD kinase (ppnk), were all shown to be upregulated by both 0.4 mM and 0.8 mM copper, except for the 5’-nucleotidase (surE) gene that was downregulated under the second treatment. Among the 11 genes associated with the metabolism of polyhydroxyalkanoates, only the 3-ketoacyl-CoA-thiolase (phaA) gene was upregulated in both treatments and highly expressed in the second treatment; the majority were downregulated and repressed. Conclusion, significance and impact of study The study demonstrated that copper induces the metabolism of polyphosphates in B. thuringiensis Y1 that serve as an alternative source of energy under copper stress. This model can be extended to the study of other species of Bacillus under environmental stress.