Growth of Pseudomonas aeruginosa LP5 on 2, 5-dicchlorobenzoate: Detection of aromatic ring hydroxylating dioxygenase (ARHDO) gene

Pseudomonas aeruginosa LP5 grew on 2, 5-dichlorobenzoate with doubling time (D) 6.64 d and mean growth rate (k) 0.104 d-1. The organism showed a prolonged lag period lasting 9 days followed by a sudden rise within 3 days (D= 1.1 d; k= 0.628 d-1) and death in less than 72 hours on 2, 6-dichlorobenzoate. Polymerase chain reaction (PCR) amplification of DNA of LP5 showed aromatic dihydroxylating (ARHDO) gene band with molecular weight corresponding to the targeted fragment (0.73 kb). The capability of LP5 on dichlorobenzoates and detection of dioxygenase genes is a validation of its versatility and potential for bioremediation

Metal biouptake by actively growing cells of metal-tolerant bacterial strains

Metal uptake potentials of Pseudomonas aeruginosa CA207Ni, Burkholderia cepacia CA96Co, Rhodococcus sp. AL03Ni, and Corynebacterium kutscheri FL108Hg were studied to determine their competence in detoxification of toxic metals during growth. Metabolism-dependent metal biouptake of the bacteria revealed appreciable uptake of the metals (57–61, 10–30, 23–60, and 10–16 mg g dw−1 of Ni2+, Cr6+, Co2+, and Cd2+, respectively) from medium, after initial drop in pH, without lag phase. The bacteria exhibited 95–100 % removal efficiency for the metals from aqueous medium as 21 (±0.8)–84 (±2.0) concentration factors of the metals were transported into the bacterial systems. Passive adsorption onto the cell surfaces occurred within 2-h contact, and afterwards, there was continuous accumulation for 12 days. Biosorption data of the bacteria were only fitted into Langmuir isotherm model when strains AL96Co, CA207Ni, and AL03Ni interacted with Ni2+, achieving maximum uptake of 9.87, 2.72, and 2.69 mg g dw−1, respectively. This study established that the actively growing bacterial strains displayed, at least, 97.0 % (±1.5) continuous active removals of metals upon adsorption. The bacteria would be good candidates for designing bioreactor useful in the detoxification campaign of heavy metal-polluted systems

Degradation of cyclohexane and cyclohexanone by Bacillus lentus strain LP32

A Gram-positive bacterium, Bacillus lentus LP32, originally isolated on the basis of its ability to utilise pyrene as sole source of carbon was found to be able to grow luxuriantly on alicyclic compounds as sole substrates. It showed poor growth on anthracene, naphthalene, 1-naphthol and phenanthrene. Growth rate on cyclohexane was 1.32 d-1, while doubling time was 0.76 d. The corresponding values for growth on cyclohexanone were 0.77 d-1 and 1.29 d, respectively. Within 10 days, the amount of cyclohexane in culture reduced from 317.62 to 102.55 mgl-1, then to 23.04 mgl-1 on day 18. On cyclohexanone, substrate concentration decreased from 287.56 mgl-1 to 101.66 mgl-1 in 10 days before declining to 24.21 mgl-1 on day 18. The rate of degradation when growing on cyclohexane was 23.50 mgl-1d-1 in the first 10 days and 9.93 mgl-1d-1 between day 10 and day 18, with 67.71% degradation in 10 days and overall percentage degradation of 92.43%. On cyclohexanone, the corresponding values were 18.59 and 9.68 mg l-1d-1 as well as 64.65 and 91.58%, respectively. This organism is a potential candidate for bioremediation purpose.Keywords: Degradation, cyclohexane, cyclohexanone, alicyclic compounds

Influence of pH, temperature and nutrient addition on the degradation of atrazine by Nocardioides spp. isolated from agricultural soil in Nigeria.

Aims: To effectively exploit the atrazine degrading capabilities of Nocardioides spp. isolated from agricultural soil samples in Nigeria and ascertain the effect of pH, temperature and nutrient addition on the degradation process. Methodology and results: Isolates were cultivated on atrazine mineral salts medium at a temperature range of 4 °C - 45 °C and a pH range of 3-10. An optimum atrazine degrading activity was observed in the isolates between temperatures of 25 °C and 37 °C and between pH 5 and 8. Different carbon sources (glycerine, glucose, chitin, cellulose and sodium citrate) and nitrogen sources (urea, biuret, cyanuric acid, potassium nitrate and ammonium chloride) were also added to the medium. The addition of carbon and nitrogen sources did not increase degradation rates although urea and glycerine repressed the degradation ability of the isolates. Statistical analyses of variance at P < 0.05 showed no significant differences in the growth and degradation rates by both bacterial isolates under these conditions. Conclusion, significance and impact study: Atrazine degradation by Nocardioides spp. is pH and temperature dependent, and requires no additional sources of carbon and nitrogen. Hence, its use in bioremediation of atrazine contaminated agricultural soil should be explored

Biodegradation of p-Chloroaniline by Bacteria Isolated from Contaminated Sites

Enrichment of water from a contaminated site in a textile industry in Ikeja resulted in the isolation of two bacteria belonging to the genera Alcaligenes and Cellulomonas. These bacteria were able to mineralize para-chloroaniline (p-chloroaniline). Time course degradation of p-chloroaniline using pure cultures of these organisms showed that p-chloroaniline supported the growth of these isolates. An initial increase in cell densities of 7.50-9.46 cfu/mL was recorded from day 0-9th day for Cellulomonas sp. while for Alcaligenes denitrificans it was 7.20-9.40 cfu/mL. After day 9, a decrease in population occurred, indicating non-availability of nutrients or toxicity of the medium. Simultaneously, a decrease in the pH, indicative of increased acidity of the medium, was also observed from the first day. The result of the GC analysis of the pure isolates on p-chloroaniline shows that 86.5% of the p-chloroaniline was degraded by the Cellulomonas sp. while 81.2% was degraded by the A. denitrificans in 30 days. These bacterial isolates utilized other hydrocarbons such as pyrene, anthracene, crude oil and chlorobenzoates as sole source of carbon and energy but not phenanthrene, naphthalene and biphenyl. The two isolates tolerated NaCl concentration of up to 5%

Effects of cadmium perturbation on the microbial community structure and heavy metal resistome of a tropical agricultural soil

The effects of cadmium (Cd) contamination on the microbial community structure, soil physicochemistry and heavy metal resistome of a tropical agricultural soil were evaluated in field-moist soil microcosms. A Cd-contaminated agricultural soil (SL5) and an untreated control (SL4) were compared over a period of 5 weeks. Analysis of the physicochemical properties and heavy metals content of the two microcosms revealed a statistically significant decrease in value of the soil physicochemical parameters (P < 0.05) and concentration of heavy metals (Cd, Pb, Cr, Zn, Fe, Cu, Se) content of the agricultural soil in SL5 microcosm. Illumina shotgun sequencing of the DNA extracted from the two microcosms showed the predominance of the phyla, classes, genera and species of Proteobacteria (37.38%), Actinobacteria (35.02%), Prevotella (6.93%), and Conexibacter woesei (8.93%) in SL4, and Proteobacteria (50.50%), Alphaproteobacteria (22.28%), Methylobacterium (9.14%), and Methylobacterium radiotolerans (12,80%) in SL5, respectively. Statistically significant (P < 0.05) difference between the metagenomes was observed at genus and species delineations. Functional annotation of the two metagenomes revealed diverse heavy metal resistome for the uptake, transport, efflux and detoxification of various heavy metals. It also revealed the exclusive detection in SL5 metagenome of members of RND (resistance nodulation division) protein czcCBA efflux system (czcA, czrA, czrB), CDF (cation diffusion facilitator) transporters (czcD), and genes for enzymes that protect the microbial cells against cadmium stress (sodA, sodB, ahpC). The results obtained in this study showed that Cd contamination significantly affects the soil microbial community structure and function, modifies the heavy metal resistome, alters the soil physicochemistry and results in massive loss of some autochthonous members of the community not adapted to the Cd stress

Biodegradation potentials of polyaromatic hydrocarbon (pyrene and phenanthrene) by Proteus mirabilis isolated from an animal charcoal polluted site

Indiscriminate disposal of animal charcoal from skin and hides cottage industries often impact the environments with toxic hydrocarbon components and thus require eco-friendly remedial strategies. A bacterial strain isolated from a site polluted with animal charcoal was characterized, identified as Proteus mirabilis 10c, and studied for ability to degrade pyrene and phenanthrene. The bacterium resisted 30 µg chloramphenicol, 10 µg ampicillin, 30 µg amoxicillin and 10 µg perfloxacin; while it utilized a number of polycyclic aromatic hydrocarbons and cinnamic acid. Specific growth rate on pyrene and phenanthrene were 0.281 d−1 and 0.276 d−1, respectively. Kinetics of degradation of pyrene was 87.92 mg l−1 in 30 days at the rate of 2.93 mg l−1 d−1, biodegradation constant at 0.073 d−1 and half-life of 9.50 d. The corresponding values for phenanthrene degradation kinetics by the bacterium were 90.12 mg l−1, 3.02 mg l−1 d−1, 0.079 d−1 and 8.77 d, respectively. Efficient degradation of crude oil (92.3%) in chemically defined medium was evident with near-disappearance of most aromatic spectra in 30 days. Considering its unique physiologies and broad specificities for aromatic and aliphatic hydrocarbons, the bacterium has potentials for decommissioning environments contaminated with toxic components of animal charcoal

Monitoring of microbial hydrocarbon remediation in the soil

Bioremediation of hydrocarbon pollutants is advantageous owing to the cost-effectiveness of the technology and the ubiquity of hydrocarbon-degrading microorganisms in the soil. Soil microbial diversity is affected by hydrocarbon perturbation, thus selective enrichment of hydrocarbon utilizers occurs. Hydrocarbons interact with the soil matrix and soil microorganisms determining the fate of the contaminants relative to their chemical nature and microbial degradative capabilities, respectively. Provided the polluted soil has requisite values for environmental factors that influence microbial activities and there are no inhibitors of microbial metabolism, there is a good chance that there will be a viable and active population of hydrocarbon-utilizing microorganisms in the soil. Microbial methods for monitoring bioremediation of hydrocarbons include chemical, biochemical and microbiological molecular indices that measure rates of microbial activities to show that in the end the target goal of pollutant reduction to a safe and permissible level has been achieved. Enumeration and characterization of hydrocarbon degraders, use of micro titer plate-based most probable number technique, community level physiological profiling, phospholipid fatty acid analysis, 16S rRNA- and other nucleic acid-based molecular fingerprinting techniques, metagenomics, microarray analysis, respirometry and gas chromatography are some of the methods employed in bio-monitoring of hydrocarbon remediation as presented in this review

Degradation of spiked pyrene and non-pyrene hydrocarbons in soil microcosms by Pseudomonas species isolated from petroleum polluted soils

The abilities of three Pseudomonads, Pseudomonas sp. strain LP1, Pseudomonas aeruginosa LP5 and P. aeruginosa LP6 to survive and enhance the degradation of pyrene and non-pyrene hydrocarbons in soil were tested in field-moist microcosms. All three organisms were able to survive and maintain high densities > × 107 in soil. In sterilized soils inoculated with bacterial isolates, 37.34%, 50.30%, and 42.21% were degraded by LP1, LP5, and LP6, respectively. The rates of pyrene degradation in soil microcosms were 0.046, 0.041, and 0.061 mg kg−1 h−1 for LP1, LP5, and LP6, respectively. A mixture of the three isolated degraded 7.73% was degraded in sterilized soil and 87.65% in native unsterilized soil (NS). The isolates also degraded non-pyrene hydrocarbon in the soils by more than 80%. The potentials these pseudomonads isolates for use as seed for bioremediation was successfully demonstrated

Equilibrium studies of cadmium biosorption by presumed non-viable bacterial strains isolated from polluted sites

Presumed non-viable high resistant Pseudomonas aeruginosa CA207Ni, Burkholderia cepacia AL96Co, Corynebacterium kutscheri FL108Hg, and Rhodococcus sp AL03Ni were studied for Cd2+ adsorption potentials. Moderate temperature, acidic pH, and high ionic strength were required for bacterial-sorption of cadmium, attaining isothermic equilibrium within 20 min. Experimental cadmium-biosorption data fitted well into biosorption isotherms. The adsorption capacities of the bacterial cell masses spanned 0.003–0.009 l mg−1 (Langmuir model) and 0.43–0.68 (Freundlich model), while binding capacity ranged from 1.14 to 56.16 mg gdw−1, with maximum achievable cadmium uptake of 62.07–109.37 mg gdw−1. The bacteria selectively removed the metal at low concentration (100.0 mg l−1) with an efficiency ranging from 50.0% to 80.0%, while approximately 80.0–92.0% removal efficiency was obtained at higher ionic concentrations (450.0 mg l−1). About 92.66% of the adsorbed metal was recovered from strain CA207Ni upon desorption, and approximately 91.7% of Cd2+ in solution was re-adsorbed onto the biomasses. In this work, effective feasible biosorption of Cd2+ in simulated wastewater system at harsh physico-chemistry, using non-viable resistant bacterial strains was demonstrated. The results indicate that the bacterial strains are sustainable tools for the detoxification of cadmium ions in industrial effluents via wastewater treatment, and cadmium demobilisation in contaminated ecosystem