Effects of carbon and nitrogen sources on rhamnolipid biosurfactant production by <Emphasis Type="Italic">Pseudomonas nitroreducens</Emphasis> isolated from soil

Rhamnolipid biosurfactant production by Pseudomonas nitroreducens isolated from petroleum-contaminated soil was investigated. The effects of carbon, nitrogen and carbon to nitrogen ratio on biosurfactant production were examined using mineral salts medium as the growth medium. The tenso-active properties (surface activity and critical micelle concentrations of the produced biosurfactant were also evaluated. The best carbon source, nitrogen source were glucose and sodium nitrate giving rhamnolipid yields of 5.28 and 4.38 g l⁻¹, respectively. The maximum rhamnolipid production of 5.46 g l⁻¹ was at C/N (glucose/sodium nitrate) of 22. The rhamnolipid biosurfactant reduced the surface tension of water from 72 to ~37 mN/m. It also has critical micelle concentration of ~28 mg l⁻¹. Thus, the results presented in our reports show that the produced rhamnolipid can find wide applications in various bioremediation activities such as enhanced oil recovery and petroleum degradation.     

Bioremediation of Soil Contaminated with Diesel Using Inorganic Nitrogen Sources: Incorporating nth-Order Algorithm in the Evaluation of Process Kinetics

In the present study, we investigated the effects of inorganic nitrogen sources—(NPK fertilizer, 15:15:15), (urea fertilizer, 46:0:0), (NH₄)₂SO₄ as well as monitored natural attenuation on the bioremediation of diesel-polluted soil. At the end of the 6-week study, the highest degradation was recorded in soil amended with NPK fertilizer (95 ± 2.77%) while the least total petroleum hydrocarbon removal was observed in monitored natural attenuation (89 ± 2.91%). Nth-order kinetics effectively described three of the treatments out of the four treatment plans. These include urea amendment (r² = 0.9925, average relative error (ARE) = 1.45%, root mean square error (RMSE) = 0.038, k_n = (3.57 ± 0.61) × 10^?2, n = 1.33), NPK fertilizer amendment (r² = 0.9751, ARE = 3.241%, RMSE = 0.086, k_n = (8.04 ± 0.23) × 10^?1, n = 0.74), and monitored natural attenuation (r² = 0.9697, ARE = 2.77%, RMSE = 0.073, k_n = (1.57 ± 0.50) × 10^?2, n = 1.16). The values of n from the nth-order kinetics parameter estimation indicated that all the treatments resulted in diesel degradation that followed a first-order kinetics path. Thus, the outcome of kinetic modeling showed that nth-order can be used as validating tool when many kinetic orders are under consideration. The phytotoxicity assay with Zea mays showed that the treatments plans resulted in germination indices of 17–55%.     

Bioremediation of Diesel-contaminated Soil by Composting with Locally Generated Bulking Agents

Herein, we conducted a study toward understanding the impact of composting of the diesel-contaminated soil with some locally available bulking agents (rice husks (RHs), sawdust (SD), and wood chips (WCs)). In order to ascertain the effectiveness of petroleum degradation by the process assayed, we compared the protocols with monitored natural attenuation (MNA). The overall degradation pattern was modeled with non-linear regression by comparing the experimental data with first and second-order kinetic equations. At the end of the six-week study, the amount of total petroleum hydrocarbon removed from contaminated soil was 98.26 ± 1.33% (amendment with SD + RHs + WCs), 96.89 ± 1.20 (RHs amendment), 96.55 ± 1.29% (amendment with SD), 90.01 ± 0.22% (WCs amendment), and 85.02 ± 0.21% (MNA). The degradation of TPH trends followed a second-order kinetic model for all the four compost treatments while the MNA was found to follow a first-order (slower) degradation pattern. In general, the results of the parameter estimate showed that amendment with mixture of the three bulking agents was 1.08 (8%) slower (k₂ = (1.289 ± 0.16) × 10^?5 (g mg^?1 d^?1), r² = 0.991) than SD amendment alone (k₂ = (1.392 ± 0.14) × 10^?5 (g mg^?1 d^?1), r² = 0.995). However, the mixture of the bulking agents was found to be 1.67, 1.41, and 2.4 times faster than amendments with WCs, rice, and MNA, respectively. The phytotoxicity test revealed that all the compost treatments except WCs resulted in germination index of ≥80% after six weeks of bioremediation tests. The outcome of the current investigation confirms the effectiveness of bulking agents (especially when combined) in the supply of nutrients for the bioremediation of diesel-impacted soil.     

Rhamnolipid biosurfactant production by Pseudomonas nitroreducens immobilized on Ca2+ alginate beads and under resting cell condition

Pseudomonas nitroreducens MILB-8054A isolated from petroleum-contaminated soil, immobilized on calcium alginate beads, and under resting cell condition, produced biosurfactants. Immobilized cells gave a best yield of 5.6 g rhamnolipid l^?1 using sucrose as carbon source. Time course study using resting cells showed that 2 % v/v of palm oil (preculture carbon source) and 10 % diesel (carbon source) gave the best rhamnolipid yield of 5.1 g l^?1 at pH 8 and temperature of 30 °C. Carbon utilization by resting cells was compared with that of growing cells. The best biosurfactant recovery procedure was acetone extraction.     

Enhancement of the Anti-inflammatory Effect of Bromelain by Its Immobilization on Probiotic Spore of Bacillus cereus

The therapeutic application of bromelain is limited due to its sensitivity to operating conditions such as high acidity, gastric proteases in the stomach juice, chemicals, organic solvents and elevated temperature. We hypothesized that bromelain immobilized on probiotic bacterial spores would show enhanced therapeutic activity through possible synergistic or additive effects. In this study, the oedema inhibition potential of bromelain immobilized on probiotic Bacillus spores was compared to the free enzyme using the carrageenan paw oedema model with Wistar rats. In batch A rats (carrageenan-induced inflammation 30 min after receiving oral treatments), group 7 rats treated with a lower dose of spore-immobilized bromelain suspension showed the highest oedema inhibition, 89.20 ± 15.30%, while group 4 treated with a lower dose of free bromelain had oedema inhibition of 60.25 ± 13.00%. For batch B rats (carrageenan-induced inflammation after receiving oral treatment for three days), group 7 rats treated with a lower dose of spore-immobilized bromelain suspension showed higher inhibition percentage (81.94 ± 8.86) than group 4 treated with a lower dose of free bromelain (78.45 ± 4.46) after 24 h. Our results showed that used alone, the enzyme and the spores produced oedema inhibition and improved the motility of the rats. The spore-immobilized bromelain formulation performed approximately 0.9-fold better than the free bromelain and the free spores at the lower evaluated dose.

     

Use of Rice Husk as Bulking Agent in Bioremediation of Automobile Gas Oil Impinged Agricultural Soil

Evaluation of rice husk (RH) as bulking agent in bioremediation of automobile gas oil (AGO) hydrocarbon polluted agricultural soil using renewal by enhanced natural attenuation (RENA) as control was the subject of the present investigation. The effect of different parameters such as total petroleum hydrocarbon (TPH), dehydrogenase activity (DHA), optical density and pH on bioremediation performance were evaluated. The studied parameters such as microbial dynamics, percentage degradation and DHA were found to be higher in RH-amended system and differed significantly with control at P < 0.05. RH resulted in high removal efficiency of 97.85 ± 0.93% under a two-month incubation period, while RENA had lesser removal efficiency of 53.15 ± 3.81%. Overall hydrocarbon biodegradation proceeded very slowly in the RENA particularly from week 0 to 4. Experimental data perfectly fitted into the first-order kinetic and generated high r² values (0.945), first-order degradation constant (0.47 day^?1), and shorter degradation half-life (1.50 d)—t_1/2 = Ln2/K and Ln2 numerically equals to 0.693 and hence written as 0.693/K. Micrococcus luteus and Rhizopus arrhizus were isolated in the present study, which displayed extreme AGO hydrocarbon biodegradative abilities. The use of RH in hydrocarbon-polluted soil significantly increased biodegradation rate and resulted in effective AGO cleanup within 2 months period. Therefore, RH provides an alternative source of bioremediation material in field application for abundant petroleum hydrocarbon soil pollution.