Biodegradation of endosulfan isomers in broth culture and soil microcosm by Pseudomonas fluorescens isolated from soil

Bacterial isolates from endosulfan-contaminated soil were grown in minimal medium and screened for endosulfan degradation. The isolate which used endosulfan and showed maximum growth was selected for detailed study. Maximum degradation in shake flask culture by Pseudomonas fluorescens was 92.80% of α and 79.35% of β endosulfan isomers in 15?days at 20?mg/L concentration, followed by 50 and 100?mg/L, while the corresponding values in static condition were 69.15 and 51.39%, respectively. Endosulfan concentration degradation declined significantly at 50 and 100?mg/L. Concomitant to degradation, release of chloride ion exhibited positive relation, while pH decreased from 7.0 to 4.53 in agitating and 7.0–5.18 in static condition. The soil microcosm study revealed maximum endosulfan degradation in sterilized soil amended with P. fluorescens. Endosulfan diol and endosulfan ether were among the products of endosulfan metabolism in broth culture, but only endosulfan ether was detected in the soil microcosm. Endosulfan sulphate, a persistent and toxic metabolite of endosulfan, was not detected in either case. The study showed that P. fluorescens could be used effectively for bioremediation of the pesticide contaminated sites.     

Peracetic acid disinfectant efficacy against Pseudomonas aeruginosa biofilms on polystyrene surfaces and comparison between methods to measure it

In food industry, biofilms are a source of recalcitrant contaminations, being possible sources of public health problems. The aims of this study were evaluate the disinfectant efficacy of peracetic acid against Pseudomonas aeruginosa biofilms grown on polystyrene surfaces, determining the best method to measuring its disinfecting efficacy. PERAsafe^® has been used as peracetic acid donor into medium. Direct colony-forming units (CFUs) counts together with two assays based in optical density measurement have been tested. Starting from bacterial suspensions (∼10⁵), the log₁₀ values obtained were 4.34 ± 0.20, 2.60 ± 0.77 and 0.00 for 0.4%, 0.8% and 1.61%, PERAsafe^® concentrations, respectively, showing that this compound achieved the 100% killing efficacy against P. aeruginosa biofilms. Comparative study among the selected tests shows similar results between CFUs counts and spectrophotometry, but there are significant differences between direct counting and McFarland test. At concentration of 1.61%, the correlation factors were 0.8998 and 0.7338 when comparing CFU direct counting with microplate absorbance measurements and McFarland turbidity tests, respectively. These results show that measurement on microplates is an effective tool to evaluate the effectiveness of PERAsafe^® against P. aeruginosa biofilms, being able to effectively replace the CFUs count in routine tests to determine the sterilization levels on surfaces.     

关于三株新血清型菌株补充绿脓杆菌国际抗原分型系统(IATS)的建议

作者在研制绿脓杆菌(PA)国际抗原分型系统(IATS)的基础上,收集了600余株绿脓杆菌,经生物学特性鉴定,交叉凝集试验和凝集素吸收试验,筛选出 CMCC-10117、PA-87、NO.446、PA-26、PS-1135株菌株(其中 NO.446、PA-26和 PS-113 3株为同型)系 IATS 不能包括的新血清型菌株。根据其抗原特异性从中选取了3株加入 IATS 中,使 IATS 由17个血清型发展为20个血清型。以新选出的MCC-10117作为 IATS-18型,PA-87作为 IATS-19型,NO.446作为 IATS-20型,经用468株临床分离的绿脓杆菌分型考核,分型率为99.4%。     

Treatment of milk by gamma irradiation—effect of anoxia on lipid peroxidation and the survival of Pseudomonas aeruginosa

One of the causes of organoleptic changes in milk, lipid peroxidation, has been monitored by a sensitive assay newly applied to foodstuffs. It has been shown that even at medium radiation doses (up to 3600 Gy) negligible lipid peroxidation takes place, providing the milk is thoroughly gassed with nitrogen. Using a test spoilage microorganism, Pseudomonas aeruginosa (Schroeter), it was shown that the D₁₀ was 35 Gy in air and 105 Gy under anaerobic conditions. Dose/survival curves indicate that 5 decades of killing needed 200 Gy in air and 600 Gy under nitrogen. However, even after receiving a dose of 1600 Gy, both aerobically and anaerobically treated milk failed the methylene blue test at 21 days post-irradiation. The data indicate that, although the deleterious lipid peroxidation effects of irradiation can be minimised by the removal of air, this in turn results in a degree of protection being afforded to the bacteria, and that doses in excess of 1600 Gy are needed to effectively sterilise milk.     

Efficiency of naphthalene biodegradation by Pseudomonas putida G7 in soil

The efficiency of naphthalene degradation by Pseudomonas putida G7 in soil was assessed using a mathematical model. The number of microorganisms and the concentration of naphthalene in soil samples were monitored. The feasibility of a spectrofluorometric method for naphthalene assay in soil samples was compared with high pressure liquid chromatography. A proposed mathematical model described the growth of the naphthalene‐degrading strains and the consumption of substrates (naphthalene, naphthalene degradation intermediates and soil organic substances) in soil. To describe the growth kinetics of microorganisms having high affinity to substrates with low solubility, two differential equations with substrate exponent 2/3 were proposed. These equations were used to describe utilization of soil organic matter. The model parameters characterize the growth rates for different substrates and respective yield coefficients, specific bacterial death and adaptation rates, and also the rates of PAHs degradation and evaporation. These characteristics can be used in choosing the bacterial strains for biopreparations and efficient clean‐up biotechnology of polluted soils. Copyright © 2004 Society of Chemical Industry     

Bioremediation of toluene‐contaminated air using an external loop airlift bioreactor

An external loop airlift bioreactor (ELAB) has been used to capture and degrade toluene from a contaminated air stream. Using a spinning sparger, the toluene could be transferred from small, uniform bubbles into the aqueous culture media with an overall mass transfer coefficient as high as 1.1 h^?1. Due to the very volatile nature of toluene, Pseudomonas putida (ATCC 23973) was cultured and maintained on benzyl alcohol, the first intermediate compound in the metabolic degradation pathway for toluene. Consequently, before successful continuous operation of the ELAB with toluene‐contaminated air, Pseudomonas putida was acclimatized to toluene by using 30 min intermittent sparging of contaminated air into the bioreactor. Continuous sparging of toluene‐contaminated air could then be successfully carried out with 100% capture and biodegradation efficiency at a contaminated air concentration of 15 mg dm^?3 and a loading rate of 35 mg dm^?3 h^?1. Higher concentrations and loading rates were only partially degraded. Although this capture matches only the low rates reported earlier using biofilters to remediate toluene, the ELAB operates using submerged culture and requires no packing which can plug during biofilter operation. In this study, Pseudomonas putida grew on toluene at a maximum specific growth rate of only 0.05 h^?1. © 2003 Society of Chemical Industry     

Oxygen diffusion through a developing biofilm of pseudomonas fluorescens

Using a specially designed miniature probe mounted into a plate across which water contaminated with the bacterium Pseudomonas fluorescens was flowing, oxygen levels at the biofilm/substrate interface have been determined. The experimental results show a rapid fall in the amount of oxygen available to the micro-organisms in the layers near the interface. As the biofilm approached 300 μm in thickness higher flow velocities across the biofilm had little effect on oxygen availability in the regions near the interface.     

Enhanced removal of selected hydrocarbons from soil by Pseudomonas aeruginosa UG2 biosurfactants and some chemical surfactants

The ability of rhamnolipid biosurfactants produced by Pseudomonas aeruginosa UG2 to wash a model hydrocarbon mixture from unsaturated soil columns was studied. Both aliphatic and aromatic hydrocarbons were effectively removed without soil clogging with non-recirculating biosurfactant solutions. Recirculation of wash solutions did not substantially affect washing efficiency. Of the several chemical surfactants tested, only Triton X-100 provided comparable hydrocarbon washing efficiency without soil clogging. The results suggest that UG2 biosurfactants have the potential for remediation of hydrophobic pollutants in unsaturated soil.     

Improved mass transfer and biodegradation rates of naphthalene particles using a novel bead mill bioreactor

One of the main challenges in the treatment of polycyclic aromatic hydrocarbons (PAHs) in controlled bioreactors is the hydrophobicity and low solubility of these compounds in the aqueous phase, resulting in appreciable mass transfer limitations within the bioreactor. To address this challenge, we have developed a modified roller bioreactor (Bead Mill Bioreactor) in which inert particles are used to improve mass transfer from the solid phase to the aqueous phase. Experimental results with naphthalene as a model PAH and Pseudomonas putida as a candidate bacterium indicate that both the mass transfer rate from the solid phase to liquid phase and the biodegradation rate in the Bead Mill Bioreactor (BMB) were significantly higher than those in a conventional roller bioreactor (20‐fold and 5.5‐fold, respectively). The enhancement of mass transfer was dependent on the type, size and volumetric loading of the inert particles, as well as concentration of particulate naphthalene. The highest mass transfer coefficient (k_La = 2.1 h^?1) was achieved with 3 mm glass beads at a volumetric loading of 50% (particle volume/working volume) with 10 000 mg dm^?3 particulate naphthalene. The maximum biodegradation rate of naphthalene attained in the bead mill bioreactor (59.2 mg dm^?3 h^?1 based on the working volume and 118.4 mg dm^?3 h^?1 based on the liquid volume) surpasses most other rates published in the literature and is equivalent to values reported for more complex bioreaction systems. The bead mill bioreactor developed in the present work not only enjoys a simple design but shows excellent performance for treatment of PAHs suspended in an aqueous phase. This fundamental information will be of significant value for future studies involving soil‐bound PAHs. Copyright © 2005 Society of Chemical Industry     

Silver nanoparticles enhance Pseudomonas aeruginosa PAO1 biofilm detachment

Objectives: Silver nanoparticles (AgNPs) with a size ranging from 7 to 70?nm were synthesized using the ascorbic acid-citrate seed-mediated growth approach at room temperature.

Methods: The 8?nm silver particles were prepared using gallic acid in alkaline conditions and used as seed to prepare AgNPs.

Results: The presence of ascorbic acid and citrate allows the regulation of size and size distribution of the nanoparticles. The increase in free silver ion-to-seed ratio (Ag⁺/Ag⁰) resulted in changes of particle shape from spherical to pseudo-spherical and minor cylindrical shape. Further, a repetitive seeding approach resulted in the formation of pseudo-spherical particles with higher polydispersity index and minor distributions of tetrahedral particles. Citrate-capped AgNPs were stable and did not agglomerate upon centrifugation. The effect of AgNPs on biofilm reduction was evaluated using static culture on 96-well microtiter plates. Results showed that AgNPs with the smallest average diameter were most effective in the reduction of Pseudomonas aeruginosa biofilm colonies, which accounted for 90% of removal.

Conclusion: The biofilm removal activities of the nanoparticles were found to be concentration-independent particularly for the concentration within the range of 80–200?µg/mL.