Screening of three phenol-degrading bacteria for copper bioremoval of copper

Aim: Potential microbial isolates for bioremoval of copper were collected from phenolic and heavy metal-contaminated soils and screened in copper-containing medium for determining the maximum tolerance level for copper. Methodology: Bioremoval of copper was assessed using sodium diethyl dithiocarbamate assay. Physical and cultural conditions influencing copper bioremoval such as initial concentration, biomass dosage (inoculum volume), temperature and pH were optimised via one-factor-at-a-time. Results: The highest maximum tolerance level was displayed by Serratia sp. AQ5-03 at 600 mg l-1, while for Alcaligenes sp. AQ5-02 and Pseudomonas sp. AQ5-04 it was 500 mg l-1. The highest bioremoval for Alcaligenes sp. AQ5-02, Serratia sp. AQ5-03 and Pseudomonas sp. AQ5-04 was achieved at 20, 50 and 75 mg l-1, respectively. The optimum biomass dosage (inoculum volume) for both Serratia sp. AQ5-03 and Pseudomonas sp. AQ5-04 were 15%, whereas it was 10% for Alcaligenes sp. AQ5-02. The results also revealed that maximum bioremoval was achieved at pH 6 at an optimum temperature of 20°C for both Alcaligenes sp. AQ5-02 and Pseudomonas sp. AQ5-04, while Serratia sp. AQ5-03 showed optimum removal at pH 7 and at 30°C temperature. The maximum bioremoval efficiency for Alcaligenes sp. AQ5-02, Serratia sp. AQ5-03 and Pseudomonas sp were found to be 62, 57 and 70%, respectively. Interpretation: The results indicate that Alcaligenes sp. AQ5-02, Serratia sp. AQ5-03, Pseudomonas sp. AQ5-04 can be utilised as bioremoval agent since it possessed the highest bioremoval efficiency for copper remediation.

Identification of Serratia marcescens SE1 and determination of its herbicide 2,2- dichloropropionate (2,2-DCP) degradation potential

Aims: The goal of the study is to isolate species of bacteria that capable of utilizing 2,2-dichloropropionic acid (2,2-DCP) as sole carbon source from soil sample collected from surrounding lake water located in Universiti Teknologi Malaysia, Skudai, Johor. Methodology and Results: Genomic DNA from bacterium SE1 was extracted and PCR amplification was carried out using universal primers, Fd1 (5’ - AGA GTT TGA TCC TGGCTC AG - 3’) and rP1 (5’- ACG GTC ATA CCT TGT TAC GAC TT - 3’) before sending for sequencing. The 16S rDNA nucleotide sequences were compared with Basic Local Alignment Search Tool nucleotide (BLASTn) and further analyzed using phylogenetic tree of Neighbour-Joining method (MEGA 5). Phylogenetic analysis indicated that SE1 strain clearly shared 97% homology to the genus of Serratia marcescens and therefore designated as Serratia marcescens sp. SE1. SE1 exhibited the ability to utilize 2,2-DCP as sole carbon source at 20 mM concentration with cell doubling time of 5 h and maximum chloride ion release of 38 μmolCl-/mL. This result suggests that the dehalogenase enzyme present in the bacteria has high affinity towards the substrate. Based on morphological and partial biochemical characteristics, strain SE1 was a non-motile Gram negative bacterium with red colonies, that gave a catalase positive reaction. Conclusion, significance and impact of study: A better understanding of dehalogenases enzyme produce by this S. marcescens sp. SE1 in general will be useful to be used as bioremediation tools for environmental management. This is the first reported case that Serratia sp. has the ability to degrade halogenated compound.

Reduction of Mo(VI) by the bacterium Serratia sp. strain DRY5.

The need to isolate efficient heavy metal reducers for cost effective bioremediation strategy have resulted in the isolation of a potent molybdenum-reducing bacterium. The isolate was tentatively identified as Serratia sp. strain DRY5 based on the Biolog GN carbon utilization profiles and partial 16S rDNA molecular phylogeny. Strain DRY5 produced 2.3 times the amount of Mo-blue than S. marcescens strain Dr.Y6, 23 times more than E. coli K12 and 7 times more than E. cloacae strain 48. Strain DRY5 required 37 oC and pH 7.0 for optimum molybdenum reduction. Carbon sources such as sucrose, maltose, glucose and glycerol, supported cellular growth and molybdate reduction after 24 hr of static incubation. The most optimum carbon source that supported reduction was sucrose at 1.0% (w/v). Ammonium sulphate, ammonium chloride, glutamic acid, cysteine, and valine supported growth and molybdate reduction with ammonium sulphate as the optimum nitrogen source at 0.2% (w/v). Molybdate reduction was optimally supported by 30 mM molybdate. The optimum concentration of phosphate for molybdate reduction was 5 mM when molybdate concentration was fixed at 30 mM and molybdate reduction was totally inhibited at 100 mM phosphate. Mo-blue produced by this strain shows a unique characteristic absorption profile with a maximum peak at 865 nm and a shoulder at 700 nm. Dialysis tubing experiment showed that 95.42% of Mo-blue was found in the dialysis tubing suggesting that the molybdate reduction seen in this bacterium was catalyzed by enzyme(s). The characteristics of isolate DRY5 suggest that it would be useful in the bioremediation of molybdenum-containing waste.

First isolation of microorganisms from the gut diverticulum of Aedes aegypti (Diptera: Culicidae): new perspectives for an insect-bacteria association

We show for the first time that the ventral diverticulum of the mosquito gut (impermeable sugar storage organ) harbors microorganisms. The gut diverticulum from newly emerged and non-fed Aedes aegypti was dissected under aseptic conditions, homogenized and plated on BHI medium. Microbial isolates were identified by sequencing of 16S rDNA for bacteria and 28S rDNA for yeast. A direct DNA extraction from Ae. aegypti gut diverticulum was also performed. The bacterial isolates were: Bacillus sp., Bacillus subtilis and Serratia sp. The latter was the predominant bacteria found in our isolations. The yeast species identified was Pichia caribbica.