Enhanced fat degradation following the addition of a Pseudomonas species to a bioaugmentation product used in grease traps

The presence of fats, oils and greases (FOGs) in wastewater can lead to many problems including blockages. Investigation of a bioaugmentation product, consisting of Bacillus spp., to degrade butter (1%, W/V) and olive oil (1%, V/V) was performed in aerobic batch cultures for 13-day incubation. Gravimetric analysis of the remaining substrates showed slowly degradation of the oil after a 2-day lag, but no degradation of the butter. Addition of a Pseudomonas putida strain CP1 to the Bacillus spp. population promoted rapidly degradation of both fats after 7?days of incubation. High fat accumulation revealed the potential use of the new bacterial mixture for production of added-value compounds. Lipase production only by the Bacillus spp. along with the analysis of the remaining lipids with thin layer chromatography and gas chromatography, suggested that the Bacillus spp. mainly only hydrolyzed the fat. The breakdown products were metabolized by the Pseudomonas putida CP1 performing preferential utilization of unsaturated fatty acids. Investigation of population dynamics using selective plating and a labeled Pseudomonas putida CP1::Tn7-gfp showed domination of the latter. The new mixture performed a successful cooperation with good potential for FOG treatment and an aggregative response desirable to fat degradation in grease traps.     

Bioremediation of Soils by Plant–Microbe Systems

Abstract

Sustainable ecosystems can be designed to eliminate environmental toxins and reduce pathogen loads through the direct and indirect consequences of plant and microbial activities. We present an approach to the bioremediation of disturbed environments, focusing on petroleum hydrocarbon (PHC) contaminants. Treatment consists of incorporating a plant-based amendment to enhance ecosystem productivity and physiochemical degradation followed by the establishment of plants to serve as oxidizers and foundations for microbial communities. Promising amendments for widespread use are entire plants of the water fern Azolla and seed meal of Brassica napus (rapeseed). An inexpensive byproduct from the manufacture of biodiesel and lubricants, rapeseed meal is high in nitrogen (6% wt/wt), stimulates >100-fold increases in populations of resident Streptomyces species, and suppresses fungal infection of roots subsequently cultivated in the amended soil. Synergistic enzymatic and chemical activities of plant and microbial metabolism in root zones transform and degrade soil contaminants. Emphasis is given to mechanisms that enable PHC functionalization via reactive molecular species.     

分子生物学在生物强化处理环境污染物中的应用

综述了分子生物学的相关技术在生物强化处理环境污染物中的应用，主要包括：通过分子生物技术育种、筛选、构建高效降解污染物微生物菌株；提供更科学、快捷、准确多样的环境监测和环境评价技术手段；探索微生物与环境污染物间的相互关系，及微生物降解污染物的有关基因定位，相关主要技术有：核酸探针检测、DNA-DNA杂交，rRNA和rRNA基因序列分析。质粒指纹图谱分析、聚合酶链式反应（PCR）、低分子量PNA分布图及由PCR技术发展而带动的基因多态性技术，如变性梯度凝胶电泳（DGGE）、随机扩增多态性DNA技术（RAPD）等。     

Quantifying chlorinated ethene degradation during reductive dechlorination at Kelly AFB using stable carbon isotopes

Stable isotope analysis of chlorinated ethene contaminants was carried out during a bioaugmentation pilot test at Kelly Air Force Base (AFB) in San Antonio Texas. In this pilot test, cis-1,2-dichloroethene (cDCE) was the primary volatile organic compound. A mixed microbial enrichment culture, KB-1, shown in laboratory experiments to reduce chlorinated ethenes to non-toxic ethene, was added to the pilot test area. Following bioaugmentation with KB-1, perchloroethene (PCE), trichloroethene (TCE) and cDCE concentrations declined, while vinyl chloride (VC) concentrations increased and subsequently decreased as ethene became the dominant transformation product. Shifts in carbon isotopic values up to 2.7 per thousand, 6.4 per thousand, 10.9 per thousand and 10.6 per thousand were observed for PCE, TCE, cDCE and VC, respectively, after bioaugmentation, consistent with the effects of biodegradation. While a rising trend of VC concentrations and the first appearance of ethene were indicative of biodegradation by 72 days post-bioaugmentation, the most compelling evidence of biodegradation was the substantial carbon isotope enrichment (2.0 per thousand to 5.0 per thousand) in ?13C(cDCE). Fractionation factors obtained in previous laboratory studies were used with isotope field measurements to estimate first-order cDCE degradation rate constants of 0.12 h(-1) and 0.17 h(-1) at 115 days post-bioaugmentation. These isotope-derived rate constants were clearly lower than, but within a factor of 2-4 of the previously published rate constant calculated in a parallel study at Kelly AFB using chlorinated ethene concentrations. Stable carbon isotopes can provide not only a sensitive means for early identification of the effects of biodegradation, but an additional means to quantify the rates of biodegradation in the field.     

Bioaugmentation and quorum sensing disruption as solutions to increase nitrate removal in sequencing batch reactors treating nitrate-rich wastewater

Bioaugmentation of denitrifying bacteria can serve as a promising technique to improve nutrient removal during wastewater treatment. While denitrification inhibition by bacterial quorum sensing (QS) in Pseudomonas aeruginosa has been indicated, the application of bacterial QS disruption to improve nitrate removal from wastewater has not been investigated. In this study, the effect of bioaugmentation of P. aeruginosa SD-1 on nitrate removal in sequencing batch reactors that treat nitrate rich wastewater was assessed. Additionally, the potential of a quorum sensing inhibitor (QSI) to improve denitrification following bacterial bioaugmentation was evaluated. Curcumin, a natural plant extract, was used as a QSI. The chemical oxygen demand (COD) and initial nitrate concentration of the influent were 700±20 mg/L and 200±10 mg/L respectively, and their respective concentrations in the effluent were 56.9±3.2 mg/L and 9.0±3.2 mg/L. Thus, the results revealed that bioaugmentation of P. aeruginosa SD-1 resulted in an increased nitrate removal to 82%±1%. Further, nitrate was almost completely removed following the addition of the QSI, and activities of nitrate reductase and nitrite reductase increased by 88%±2% and 74%±2% respectively. The nitrogen mass balance indicated that aerobic denitrification was employed as the main pathway for nitrogen removal in the reactors. The results imply that bioaugmentation and modulation of QS in denitrifying bacteria, through the use of a QSI, can enhance nitrate removal during wastewater treatment.     

Bioaugmentation with butane-utilizing microorganisms to promote in situ cometabolic treatment of 1,1,1-trichloroethane and 1,1-dichloroethene

A field study was performed to evaluate the potential for in-situ aerobic cometabolism of 1,1,1-trichloroethane (1,1,1-TCA) through bioaugmentation with a butane enrichment culture containing predominantly two Rhodococcus sp. strains named 179BP and 183BP that could cometabolize 1,1,1-TCA and 1,1-dicholoroethene (1,1-DCE). Batch tests indicated that 1,1-DCE was more rapidly transformed than 1,1,1-TCA by both strains with 183BP being the most effective organism. This second in a series of bioaugmentation field studies was conducted in the saturated zone at the Moffett Field In Situ Test Facility in California. In the previous test, bioaugmentation with an enrichment culture containing the 183BP strain achieved short term in situ treatment of 1,1-DCE, 1,1,1-TCA, and 1,1-dichloroethane (1,1-DCA). However, transformation activity towards 1,1,1-TCA was lost over the course of the study. The goal of this second study was to determine if more effective and long-term treatment of 1,1,1-TCA could be achieved through bioaugmentation with a highly enriched culture containing 179BP and 183BP strains. Upon bioaugmentation and continuous addition of butane and dissolved oxygen and or hydrogen peroxide as sources of dissolved oxygen, about 70% removal of 1,1,1-TCA was initially achieved. 1,1-DCE that was present as a trace contaminant was also effectively removed ( 80%). No removal of 1,1,1-TCA resulted in a control test leg that was not bioaugmented, although butane and oxygen consumption by the indigenous populations was similar to that in the bioaugmented test leg. However, with prolonged treatment, removal of 1,1,1-TCA in the bioaugmented leg decreased to about 50 to 60%. Hydrogen pexoxide (H₂O₂) injection increased dissolved oxygen concentration, thus permitting more butane addition into the test zone, but more effective 1,1,1-TCA treatment did not result. The results showed bioaugmentation with the enrichment cultures was effective in enhancing the cometabolic treatment of 1,1,1-TCA and low concentrations of 1,1-DCE over the entire period of the 50-day test. Compared to the first season of testing, cometabolic treatment of 1,1,1-TCA was not lost. The better performance achieved in the second season of testing may be attributed to less 1,1-DCE transformation product toxicity, more effective addition of butane, and bioaugmentation with the highly enriched dual culture.     

Bioaugmentation of a 4-chloronitrobenzene contaminated soil with Pseudomonas putida ZWL73

The strain Pseudomonas putida ZWL73, which metabolizes 4-chloronitrobenzene (4CNB) by a partial-reductive pathway, was inoculated into lab-scale 4CNB-contaminated soil for bioaugmentation purposes in this study. The degradation of 4CNB was clearly stimulated, as indicated with the gradual accumulation of ammonium and chloride. Simultaneously, the diversity and quantity of cultivable heterotrophic bacteria decreased due to 4CNB contamination, while the quantity of 4CNB-resistant bacteria increased. During the bioaugmentation, denaturing gradient gel electrophoresis analysis showed the changes of diversity in dominant populations of intrinsic soil microbiota. The results showed that Alphaproteobacteria and Betaproteobacteria were not distinctly affected, but Actinobacteria were apparently stimulated. In addition, an interesting dynamic within Acidobacteria was observed, as well as an influence on ammonia-oxidizing bacteria population. These combined findings demonstrate that the removal of 4CNB in soils by inoculating strain ZWL73 is feasible, and that specific populations in soils rapidly changed in response to 4CNB contamination and subsequent bioaugmentation.     

Performance of bioaugmentation-assisted phytoextraction applied to metal contaminated soils: a review

Bioaugmentation-assisted phytoextraction is a promising method for the cleaning-up of soils contaminated by metals. Bacteria mainly Plant Growth Promoting Rhizobacteria (PGPR) and fungi mainly Arbuscular Mycorrhizal Fungi (AMF) associated with hyperaccumulating or non-hyperaccumulating plants were analyzed on the basis of a bioprocess engineering approach (concentration and amount of metals extracted by plants, translocation and bioconcentration factor, and plant biomass). In average bioaugmentation increased metals accumulated by shoots by a factor of about 2 (metal concentration) and 5 (amount) without any obvious differences between bacteria and fungi. To optimize this process, new relevant microorganism-plant associations and field scale experiments are needed along with a common methodology for the comparison of all experiments on the same basis. Recommendations were suggested concerning both the microbial-plant selection and the implementation of bioaugmentation to enhance the microbial survival. The use of microbial consortia associated with plant was discussed notably for multi-contaminated soils.     

Rotating biological contactors for wastewater treatment – A review

Rotating biological contactors (RBCs) for wastewater treatment began in the 1970s. Removal of organic matter has been targeted within organic loading rates of up to 120 g m⁻² d⁻¹ with an optimum at around 15 g m⁻² d⁻¹ for combined BOD and ammonia removal. Full nitrification is achievable under appropriate process conditions with oxidation rates of up to 6 g m⁻² d⁻¹ reported for municipal wastewater. The RBC process has been adapted for denitrification with reported removal rates of up to 14 g m⁻² d⁻¹ with nitrogen rich wastewaters. Different media types can be used to improve organic/nitrogen loading rates through selecting for different bacterial groups. The RBC has been applied with only limited success for enhanced biological phosphorus removal and attained up to 70% total phosphorus removal. Compared to other biofilm processes, RBCs had 35% lower energy costs than trickling filters but higher demand than wetland systems. However, the land footprint for the same treatment is lower than these alternatives. The RBC process has been used for removal of priority pollutants such as pharmaceuticals and personal care products. The RBC system has been shown to eliminate 99% of faecal coliforms and the majority of other wastewater pathogens. Novel RBC reactors include systems for energy generation such as algae, methane production and microbial fuel cells for direct current generation. Issues such as scale up remain challenging for the future application of RBC technology and topics such as phosphorus removal and denitrification still require further research. High volumetric removal rate, solids retention, low footprint, hydraulic residence times are characteristics of RBCs. The RBC is therefore an ideal candidate for hybrid processes for upgrading works maximising efficiency of existing infrastructure and minimising energy consumption for nutrient removal. This review will provide a link between disciplines and discuss recent developments in RBC research and comparison of recent process designs are provided (Section 2). The microbial features of the RBC biofilm are highlighted (Section 3) and topics such as biological nitrogen removal and priority pollutant remediation are discussed (Sections 4 Biological nutrient removal in RBCs, 5 Priority pollutant remediation in RBCs). Developments in kinetics and modelling are highlighted (Section 6) and future research themes are mentioned.     

表面活性剂淋洗与投菌法联合修复2,4,6-三氯苯酚污染土壤

为了解决高浓度氯酚土壤污染问题,提出了一种表面活性剂淋洗与投菌法相结合的土壤修复方法。该方法采用表面活性剂淋洗污染土壤,使高浓度氯酚得到大部分去除后再加入微生物降解菌,从而实现低浓度氯酚的持续降解。通过2,4,6-三氯苯酚(2,4,6-TCP)的增溶实验、吸附实验以及表面活性剂的酶抑制实验,对蔗糖酯(SE)、鼠李糖脂(RL)、茶皂素(TS)、脂肪醇聚氧乙烯醚(AEO-9)4种表面活性剂进行了筛选。结果表明,SE增溶、解吸的综合效果较好,且具备良好的生物相容性,适用于淋洗有机污染土壤。淋洗-投菌联合实验表明,以0.5wt%蔗糖酯水溶液作为淋洗剂淋洗土壤并投菌降解,25 d内土壤中的2,4,6-三氯苯酚从190.4降至3.1 mg/kg,去除率达到98.4%。