A comprehensive case study was undertaken at the Blue Plains wastewater treatment plant (WWTP) to explore the bioaugmentation technique of introducing nitrifying sludge into the non-nitrifying stage over the course of two operational years. This innovative approach involved the return of waste activated sludge (WAS) from the biological nutrient removal (BNR) system to enhance the nitrification in the high carbon removal rate system. The complete ammonia oxidizer (comammox) Nitrospira Nitrosa was identified as the main nitrifier in the system. Bioaugmentation was shown to be successful as nitrifiers returned from BNR were able to increase the nitrifying activity of the high carbon removal rate system. There was a positive correlation between returned sludge from the BNR stage and the specific total kjeldahl nitrogen (TKN) removal rate in A stage. The bioaugmentation process resulted in a remarkable threefold increase in the specific TKN removal rate within the A stage. Result suggested that recycling of WAS is a simple technique to bio-augment a low SRT system with nitrifiers and add ammonia oxidation to a previously non-nitrifying stage. The results from this case study hold the potential for applicable implications for other WWTPs that have a similar operational scheme to Blue Plains, allowing them to reuse WAS from the B stage, previously considered waste, to enhance nitrification and thus improving overall nitrogen removal performance. PRACTITIONER POINTS: Comammox identifying as main nitrifier in the B stage. Comammox enriched sludge from B stage successfully bio-augmented the East side of A stage up to threefold. Bioaugmentation of comammox in the West side of A stage was potentially inhibited by the gravity thickened overflow. Sludge returned from B stage to A stage can improve nitrification with a very minor retrofits and short startup times.

This study aims at the application of a marine fungal consortium (Aspergillus sclerotiorum CRM 348 and Cryptococcus laurentii CRM 707) for the bioremediation of diesel oil-contaminated soil under microcosm conditions. The impact of biostimulation (BS) and/or bioaugmentation (BA) treatments on diesel-oil biodegradation, soil quality, and the structure of the microbial community were studied. The use of the fungal consortium together with nutrients (BA/BS) resulted in a TPH (Total Petroleum Hydrocarbon) degradation 42% higher than that obtained by natural attenuation (NA) within 120 days. For the same period, a 72 to 92% removal of short-chain alkanes (C12 to C19) was obtained by BA/BS, while only 3 to 65% removal was achieved by NA. BA/BS also showed high degradation efficiency of long-chain alkanes (C20 to C24) at 120 days, reaching 90 and 92% of degradation of icosane and heneicosane, respectively. In contrast, an increase in the levels of cyclosiloxanes (characterized as bacterial bioemulsifiers and biosurfactants) was observed in the soil treated by the consortium. Conversely, the NA presented a maximum of 37% of degradation of these alkane fractions. The 5-ringed PAH benzo(a)pyrene, was removed significantly better with the BA/BS treatment than with the NA (48 vs. 38 % of biodegradation, respectively). Metabarcoding analysis revealed that BA/BS caused a decrease in the soil microbial diversity with a concomitant increase in the abundance of specific microbial groups, including hydrocarbon-degrading (bacteria and fungi) and also an enhancement in soil microbial activity. Our results highlight the great potential of this consortium for soil treatment after diesel spills, as well as the relevance of the massive sequencing, enzymatic, microbiological and GC-HRMS analyses for a better understanding of diesel bioremediation.

Oily sludge is a residue from the petroleum industry composed of a mixture of sand, water, metals, and high content of hydrocarbons (HCs). The heavy oily sludge used in this study originated from Colombian crude oil with high density and low American Petroleum Institute (API) gravity. The residual waste from heavy oil processing was subject to thermal and centrifugal extraction, resulting in heavy oily sludge with very high density and viscosity. Biodegradation of the total petroleum hydrocarbons (TPH) was tested in microcosms using several bioremediation approaches, including: biostimulation with bulking agents and nutrients, the surfactant Tween 80, and bioaugmentation. Select HC degrading bacteria were isolated based on their ability to grow and produce clear zones on different HCs. Degradation of TPH in the microcosms was monitored gravimetrically and with gas chromatography (GC). The TPH removal in all treatments ranged between 2 and 67%, regardless of the addition of microbial consortiums, amendments, or surfactants within the tested parameters. The results of this study demonstrated that bioremediation of heavy oily sludge presents greater challenges to achieve regulatory requirements. Additional physicochemical treatments analysis to remediate this recalcitrant material may be required to achieve a desirable degradation rate.

Polycyclic Aromatic Hydrocarbons (PAHs) are major toxic and recalcitrant pollutants in the environment. This study assessed the capacity of an isolated soil microbial consortium (OMC) to biodegrade PAHs. OMC was able to reach 100% biodegradation of naphthalene, acenaphthylene, acenaphthene, fluorene and phenanthrene in solution, and up to 76% and 50% of anthracene and fluoranthene, respectively, from a mix of 16 PAHs. To measure phenanthrene (PHE) mineralization, OMC and eight strains isolated from OMC were used and identified by PCR amplification of the gene 16S ribosomal RNA. A novel Stenotrophomonas maltophilia CPHE1, not previously described as a PAH degrader, was able to mineralize almost 40% PHE and biodegrade 90.5% in solution, in comparison to OMC that reached 100% PHE degradation, but only 18.8% mineralization. Based on metabolites identified during PHE degradation and on the detection of two genes (PAH RHDα and nahAc) in OMC consortium, two possible via were described for its degradation, through salicylic and phthalic acid. PAH RHDα, which codified the first step on PHE biodegradation pathway, was also found in the DNA of S. maltophilia CPHE1. An ecotoxicology study showed that PHE bioremediation after inoculating S. maltophilia CPHE1 for 30 days decreased by half the solution toxicity.

In present study, chitinolytic bacteria were employed to bioaugment the biogas production from fungal pretreated agricultural residues. The fungal pretreatment of wheat and pearl millet straw was done by Chaetomium globosporum. Pretreated straw were digested anaerobically at batch scale with and without the presence of chitinolytic bacteria. Contrary to untreated samples, the addition of chitionolytic bacteria with pretreated wheat and pearl millet straw provided 41 and 57% higher biogas yield. The study was further upscaled to continuous stirred tank reactors. At continuous scale too, wheat straw pretreated with Chaetomium globosporum combined with chitinolytic bacteria resulted in 16% higher biogas yield in contrast to untreated straw. Higher abundance of methanogens was detected in reactors running with pretreated wheat straw during microbial community analysis. The identified bacteria belonged mostly to Firmicutes, Bacteroidetes, Proteobacteria phyla.

The present study explored bacterial aerobic biodegradation of reduced carbon-contaminants (RCC) in a pilot system mimicking remediation of a saturated aquifer in a permeable reactive biobarrier (PRBB). Bioaugmentation was performed with a pure culture of Pseudomonas putida macro-encapsulated in a cellulose-acetate porous envelope and integrated transversely to the flow trajectory of the fluid in the biobarrier and compared with controls without capsules. The macro-encapsulation technique applied allowed the incorporation of a built-in nutrient core for the slow release of macronutrients, i.e. N, P, instead of exogenous nutrients supply. 3-Chlorophenol (3CP) at a concentration range of 350-500 mg/L was chosen as an RCC model compound. The findings indicate efficient 3CP biodegradation during the PRBB operation with a similar degree of transformation (76 ± 2% and 72 ± 2%) and mineralization (55 ± 4% vs. 49 ± 3%) for exogenous and built-in nutrients supply, respectively. The extent of dechlorination in both cases (54 ± 10% vs. 40 ± 2%, respectively) followed mineralization rather than transformation, suggesting that Cl- release took place in late transformation stages. Negligible decontamination was observed in the control system without bioaugmentation. Concluding, tailored PRBB with macro-capsules incorporating a built-in nutrient core to support bacterial growth presents a significant environmental advantage controlling excess nutrients release required in bioremediation of oligotrophic systems.

Bioremediation of polycyclic aromatic hydrocarbons (PAHs) in soil matrix is often arbitrated by the presence of soil organic matter (SOM). The present study focuses on determining the effect of the soil organic content on the mycoremediation of two model PAHs, phenanthrene (Phe) and benzo(a) pyrene (BaP) through microcosm studies. On comparing various microcosm strategies, a maximum degradation of Phe (99%) and BaP (48.5%) in soil bioaugmented with Pleurotus dryinus IBB 903, followed by, biostimulation with the degradation of Phe (89.9%) and BaP (24.8%) were noted. On relating the degradation pattern with lignolytic enzyme cocktail production, a laccase activity of 108 U/Kg at day 21, aryl alcohol oxidase (411 U/Kg) and manganese peroxidase (52.2 U/Kg) at day 14 along with lignin peroxidase (481 U/Kg) at day 21 were noted in fungal augmented soils, which were comparatively higher than levels observed in the bio-stimulation. Investigating the impact of different concentration of SOM (3-12%), a maximum remediation of Phe by 100% at 9% SOM in days 28 and 58.19% for BaP at 12% SOM, respectively was exhibited. Further, the biosorption effect of PAHs in abiotic condition showed a positive correlation with the increase in SOM, with a maximum adsorption of 3.78% Phe, and 6.93% BaP. The results support that the nominal adsorption ability of SOM, and helps in enhancing the microbial growth, thereby improving their degradation potentials, when less than 6% of SOM was utilized. Overall, this work establishes the critical role of organic matter in the soil with reference by simultaneous stimulation and degradation capability in complete PAHs remediation.

A consortium of highly degrading microorganisms was used in an integrated bioaugmentation/electrocoagulation process for treating olive mill wastewater. The system was investigated for treating 1 m3 day-1, at a pilot scale, for 2 years; hydraulic loading rate and organic loading rate were 2880 l m-2 day-1 and 37,930 g COD m-2 day-1, respectively. Average removal efficiency for COD, oils, and total phenols was 63.9%, 85.2%, and 43.6%, respectively. The olive mill consortium, OMC, consisted of seven actinomycete strains. The strains were confirmed, by 16S rDNA analysis, to belong to five Streptomyces, one Kitasatospora, and one Micromonospora strains, at 100-99.06% similarities. Hydrolytic enzyme activities of OMC strains were remarkably higher for degrading cellulosic and lipid constituents (enzyme-cumulative indices, 14-16.1), than the phenolic constituents (indices, 4.1-6.5). The establishment of actinomycetes in the treatment system was indicated by their increased counts in the biofilm at the end of the biofilter, reaching 13-fold higher than that in the control bed. The treated effluent was toxic to the seedlings of Jatropha curcas (Jatropha) and Simmondsia chinensis (Jojoba). Though its application in irrigation of 3-year-old Jatropha shrubs, significantly, enhanced the fruit yield up to 1.85-fold higher than the control, without affecting the seed oil content, after 3-month application, the irrigated soil showed insignificant changes in its biochemical properties. This developed bioaugmentation/electrocoagulation process can treat wastewater with extremely high organic strength, while its approximate construction and operational costs are limited to 0.03 and 0.51 US$ m-3, respectively. It produces a treated effluent that can be reused in irrigation of specific plants. Graphical abstract.

Deep treatment is a common approach to enhance pollutant removal for biological wastewater treatment technologies (BWTTs), and life cycle assessment (LCA) holds substantial advantages to support process optimization. However, there lacks of LCA-based benchmarks that cover human-nature nexuses and stakeholder involvement, which limits the guidance and eco-design of BWTTs. This study proposed a decision-support system (DSS) by linking LCA with Water Quality Model and Conjoint Analysis. Three major findings were identified based on a demonstrative case (constructed wetland bioaugmented by dosing different microbial inocula): (1) Increasing bacterial intensities would achieve net environmental improvement, but it might not apply to all cases; (2) Making full use of natural self-purification capacity could partly replace the functions of BWTTs; (3) Stakeholders would concern aquatic environmental improvement when receiving river that had limited environmental capacity. Overall, the DSS provided a data-driven platform for screening options before determinations were made to constrain wastewater treatment sustainability.

A 90 days greenhouse experiment was conducted for evaluation of soil microbial diversity in different treatments of rhizospheric and nonrhizospheric oil sludge contaminated soil. Various pot treatments (T1-T5) were as follows: 2% oil sludge contaminated soil was considered as control (T1); augmentation of control with preadapted microbial consortium was T2; addition of Vetiver zizanioide to control was T3; bioaugmentation of control along with V. zizanioide was T4; and bioaugmentation with V. zizanioide and bulking agent was T5. During the study, different microbial populations were determined in all treatments. Additionally, soil microbial diversity using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of 16S rDNA was carried out. At the end of experimental period, significant increase in microbial number in bioaugmented rhizospheric treatments (T4 and T5) was observed as compared to non-rhizospheric and non-bioaugmented treatments (T2 and T3). The community and sequencing results revealed that combined treatment of plant and microbes resulted in improved microbial species and number. The dominant phyla belonged to γ proteobacteria, β proteobacteria, Chloroflexi, firmicutes, and uncultured bacteria. It is concluded that plant-microbe-soil system supports immense oil degrading microbial diversity and can be used as an effective indicator tool for remediation of oil sludge contaminated sites.