ACADEMIA Letters

Green energy through synergistic integration of

wastewater and algae
Jason Peterson, Utah State University
Ronald Sims, Utah State University

Abstract
Green energy can come from renewable sources that includes wastewaters from industries
mixed with nuisance microalgae biomass harvested from open ponds, lagoons, and/or harm-
ful algal blooms for co-digestion at water resource recovery facilities (WRRF). Microalgae,
however, present challenges related to biodegradability and ammonia toxicity. The research
described here investigated the bioaugmentation of waste-microalgae mixtures with biosolids
exhibiting algae-degrading activity compared with the use of the non-acclimated biosolids
used in most conventional systems of wastewater treatment. Industry wastewaters tested were
sourced from dairy, swine, and petrochemical industries. Our results showed statistically sig-
nificant advantages of the algae-degrading biosolids compared with the conventional biosolids
for bioenergy production and treatment effectiveness. This research has the potential to mit-
igate the negative impacts of nuisance algae in the environment while providing a source for
producing green energy in a “waste-to-value” model for industry wastes.

Introduction
As a source of alternative energy, green energy comes from renewable sources that include
wastewaters from industries mixed with microalgae biomass for co-digestion at water resource

Academia Letters, October 2021 ©2021 by the authors — Open Access — Distributed under CC BY 4.0

Corresponding Author: Ronald Sims, ron.sims@usu.edu

Citation: Peterson, J., Sims, R. (2021). Green energy through synergistic integration of wastewater and algae.
Academia Letters, Article 3732. https://doi.org/10.20935/AL3732.

1
recovery facilities (WRRF). Microalgae presence in open ponds and lagoons, as well as occur-
rence as harmful algal blooms are a current and increasing threat to the aquatic environment
and human health. One challenge with digestion of microalgae to biomethane is the resistance
to initial biodegradation of microalgae cells (Cho et al., 2013; Schwede et al., 2013; Ward et
al, 2014; Doloman, et al., 2019). Another challenge is the toxic inhibition of microalgae bio-
transformation by ammonia released in the anaerobic digestion process (Sievers and Brune,
1978; Yen and Brune, 2007). Successful anaerobic digestion of microalgae includes the need
to balance the carbon to nitrogen (C:N) ratio, with optimal mass ratios reported to be between
20:1 and 30:1 (Wang, X. et al., 2014; Sievers and Brune, 1978, Ward et al;l, 2008; and Es-
posito et al. 2012). Anaerobic digestion for biomethane production might also be improved
through the addition of a microbial consortium acclimated to microalgae and demonstrated to
exhibit algalytic activity (Doloman, et al., 2019).
This research describes the bioaugmentation of industry waste-microalgae mixtures with
sediment biosolids from a facultative wastewater lagoon supporting the growth of algae and
demonstrated to exhibit algalytic activity (Doloman, et al., 2017, 2019; Soboh et al. 2016).
Biomethane production was compared with the same mixtures using conventional anaerobic
digester solids taken from a conventional activated sludge WRRF.

Materials and Methods

Wastewater and microalgae sources. Three industry wastewater sources were tested for co-
digestion with microalgae that was utilized to establish a C:N mass ratio of 25:1 (Soboh,
et al., 2016). Dairy wastewater was obtained from the Utah State University Caine Dairy
Research Center. Swine wastewater was obtained from Circle Four Farms, southern Utah.
Petroleum wastewater was obtained from a local refining industry located in Woods Cross,
Utah. The mixed-culture microalgae biomass mixed was harvested from a facultative la-
goon (460 acres) at the Logan, Utah, wastewater lagoons facility that treats 12 million gallons
per day of municipal wastewater. Major microalgae occurring in the mixed culture include
Chlorella, Scenedesmus, cyanobacteria, Pediastrum, and diatoms (Christenson and Sims,
2012). Biosolids at the bottom depth of 2.3 m. within the lagoon had been exposed to sed-
imenting microalgae for a period of 52 years, and were shown to contain algalytic bacterial
species of Pseudomonas, Alcaligenes, and Citrobacter as described in Doloman et al. (2017
and 2019). The Central Weber WRRF that treats 12 MGD municipal wastewater, Ogden,
Utah, was used as the source of conventional anaerobic sludge from the anaerobic digester
and provided the inoculum matrix and microbes for biomethane production in control sys-
tems. The performance of the facultative lagoon sediment was compared with the WRRF

Academia Letters, October 2021 ©2021 by the authors — Open Access — Distributed under CC BY 4.0

Corresponding Author: Ronald Sims, ron.sims@usu.edu

Citation: Peterson, J., Sims, R. (2021). Green energy through synergistic integration of wastewater and algae.
Academia Letters, Article 3732. https://doi.org/10.20935/AL3732.

2
anaerobic sludge.
Laboratory study design. Laboratory-scale (500 mL flasks with 350 mL working volume)
anaerobic digestion systems were inoculated with either the lagoon bottom solids or the con-
ventional anaerobic digester solids. A biomethane potential (BMP) test was used to quantify
biogas production using a manometer method that measured the amount (volume) of water
displaced by the biogas and correlating pressure of the system to barometric pressure (Peter-
son, J., 2017; Esposito et al., 2012; Browne et al., 2013). Treatments were mixed on a stir
plate, and made anaerobic by bubbling nitrogen through the wastewater while monitoring the
dissolved oxygen. Temperature was maintained at 30 degrees C in an environmental control
chamber. The use of distilled water as a control for wastewater allowed for evaluation of the
effects of inoculum matrix on biogas production.
Biogas components analysis. The composition of the biogas, including methane and car-
bon dioxide, was determined weekly over the 60-day duration of the test using gas chromatog-
raphy. The biogas was analyzed using the Agilent 7890B Gas Chromatograph with a thermo-
conductivity detector and a pneumatic sampling valve. Ultra high purity helium was used as
a carrier gas set at a constant pressure of 20 PSI. Contents of samples were separated using a
HP Gas Pro column. Temperature of column was maintained at 25 degrees C in an isothermal
oven. The Analysis of biogas through gas chromatography allowed for separation of nitrogen,
methane, and carbon dioxide to determine the percentage of each gas.
Statistical analysis of results. Triplicate treatments were evaluated for determination of av-
erage and standard deviation (S.D.) values for biomethane production for each waste/inoculum
matrix combination. To determine if one inoculum treatment was statistically significantly
better, treatments were compared using single and two factor ANOVA tables. Treatments
were grouped based on inoculant and wastewater type, and a significant difference was deter-
mined based on a 0.05 alpha level.

Results and Discussion

Comparison across wastes. Results of BMP testing of the three industry wastes inoculated
with either facultative lagoon sediment (FLS) or anaerobic digester solids (ADS) demon-
strated that both systems produced comparable total biogas (methane + carbon dioxide) vol-
umes (approximately 3,000 ml) over the 60-day test duration. The FLS inoculated wastes
demonstrated less variation (91 ml S.D. for all three wastes combined) than the ADS inocu-
lated wastes (270 ml S.D.). Also, FLS treated wastes demonstrated a shorter lag time (less
than 10 days) in biogas production and also sustained biogas production for a longer time
(approximately 50 days versus 32 days).

Academia Letters, October 2021 ©2021 by the authors — Open Access — Distributed under CC BY 4.0

Corresponding Author: Ronald Sims, ron.sims@usu.edu

Citation: Peterson, J., Sims, R. (2021). Green energy through synergistic integration of wastewater and algae.
Academia Letters, Article 3732. https://doi.org/10.20935/AL3732.

3
 The FLS inoculated wastes demonstrated a significant advantage over the ADS inoculated
wastes with regard to the CH4:CO2 Ratio (vol/vol) of biogas produced. FLS treatment results
demonstrated ratios of 7.4, 10.7, and 8.8 for dairy, swine, and petrochemical wastes, while
ADS results demonstrated ratios of 6.5, 9.5, and 7.4, respectively, with all differences being
statistically significant. Figure 1 shows the CH4:CO2 ratios for the three industry wastes and
also municipal waste and the statistically averaged values for all wastes in the “Combined”
category.

Figure 1. Methane:Carbon Dioxide Ratios (ml/ml) comparing Facultative Lagoon Sludge

(FLS) with Anaerobic Digester Sludge (ADS) for treatment of microalgae mixed with wastes.
Municipal waste was used as a control and “Combined” represented the statistical average of
the four wastes identified.
The FLS inoculated wastes also demonstrated an increased mass of CH4 production per
mass of volatile solids (mg/gm). FLS treatment results demonstrated values of 308, 272, and

Academia Letters, October 2021 ©2021 by the authors — Open Access — Distributed under CC BY 4.0

Corresponding Author: Ronald Sims, ron.sims@usu.edu

Citation: Peterson, J., Sims, R. (2021). Green energy through synergistic integration of wastewater and algae.
Academia Letters, Article 3732. https://doi.org/10.20935/AL3732.

4
327 mg/gm, while the ADS treatment results demonstrated values of 240, 220, and 230 for
dairy, swine, and petrochemical wastes, respectively.
Results for each waste. For the dairy waste, the FLS treatment also showed statistically
significantly better performance for measurements including rate and duration (41 days) of
biogas production, maximum methane concentration (81%), and decrease in BOD and COD.
ADS treatment showed statistically better performance for measurements including maximum
rate of biogas production and total decrease in volatile solids.
For the swine waste, the FLS treatment also showed statistically significantly better per-
formance in total biogas (3,100) and duration of biogas production, as well as decrease in
BOD. ADS treatment showed statistically better performance for decrease in volatile solids.
For the petrochemical waste, the FLS treatment also showed statistically significantly bet-
ter performance in the duration of the average rate of biogas production and decrease in BOD.
ADS treatment showed statistically significantly better performance for measurements includ-
ing maximum rate of biogas production (120 versus 94 ml/day) and reduction in volatile solids.
Results for controls. Results with FLS and ADS inoculants alone without any wastewater
produced minimal biogas over the 60 days of the BMP test. FLS demonstrated 18 ml total
biogas and ADS demonstrated 49 ml total biogas. Algae addition alone to the wastewaters
added approximately 500 ml of total biogas over the 60-day test period, which accounted for
approximately 16% of the total biogas production of approximately 3,000 ml.
Applications of research. Applications of the research reported here include assessing
the potential for adding microalgae harvested from harmful algal blooms in Utah lakes and
reservoirs to existing anaerobic digesters at the Central Weber Facility and other WRRF for
generating additional bioenergy. Another application is the potential for seeding a proposed
new anaerobic digester at the Logan Water Reclamation Facility with lagoon sediment from
the Logan Lagoons for transforming the microalgae in the lagoons into bioenergy for the
Logan Water Reclamation Facility operations.

Conclusions
All three industry wastewater sources, including dairy, swine, and petrochemical produced
biogas with a high percentage of methane when bioaugmented with facultative lagoon sed-
iment containing algalytic activity or with conventional anaerobic digester sludge. Results
indicated the potential for the recycling of waste carbon into methane for water reclamation
facilities receiving wastes from different sources while simultaneously potentially treating
microalgae harvested from lakes as harmful algal blooms (HABs) or wastewater lagoon mi-
croalgae.

Academia Letters, October 2021 ©2021 by the authors — Open Access — Distributed under CC BY 4.0

Corresponding Author: Ronald Sims, ron.sims@usu.edu

Citation: Peterson, J., Sims, R. (2021). Green energy through synergistic integration of wastewater and algae.
Academia Letters, Article 3732. https://doi.org/10.20935/AL3732.

5
Funding and facilities support
This research was supported by the Logan, Utah, Environmental Department (Mr. Issa Hamud)
and the Central Weber Water Reclamation Facility (Mr. Kevin Hall), by the Huntsman En-
vironmental Research Center at Utah State University, and by the Sustainable Waste to Bio-
products Engineering Center at Utah State University.

Conflict of interest
The authors declare that there are no conflicts of interest.

Academia Letters, October 2021 ©2021 by the authors — Open Access — Distributed under CC BY 4.0

Corresponding Author: Ronald Sims, ron.sims@usu.edu

Citation: Peterson, J., Sims, R. (2021). Green energy through synergistic integration of wastewater and algae.
Academia Letters, Article 3732. https://doi.org/10.20935/AL3732.

6
References
Browne, J.D., Allen, E., Murphy, J.D., 2013. Evaluation of the biomethane potential from
multiple waste streams for a proposed community scale anaerobic digester. Environ. Tech-
nol. 34 (13-14), 2027-2038.

Cho, S., Park, S., Seon, J., Yu, J., Lee, T., 2013. Evaluation of thermal, ultrasonic and alkali
pretreatments on mixed-microalgal biomass to enhance anaerobic methane production.
Biores. Technol. 143, 330-336.

Christenson, L., and Sims, R.C., 2012. Rotating algal biofilm reactor and spool harvester for
wastewater treatment with biofuels by-products. Biotechnol. Bioeng. 109: 1674-1684.

Doloman, A.; Soboh, Y.; Walters, A.J.; Sims, R.C.; Miller, C.D. 2017. Qualitative Analysis
of Microbial Dynamics during Anaerobic Digestion of Microalgal Biomass in a UASB
Reactor. Int. J. Microbiol. 5291283.

Doloman, A., Pererva, Y., Cortex, M., Sims, R.C., and Miller, C.D. 2019. Augmentation
of granular anaerobic sludge with algalytic bacteria enhances methane production from
microalgal biomass. Fermentation 5(4), 88.

Esposito, G., Frunzo, L., Liotta, F., Panico, A., Pirozzi, F., 2012. Bio-methane potential tests
to measure the biogas production from the digestion and co-digestion of complex organic
substrates. The Open Environ. Eng. Jour. 5 (1).

Peterson, Jason. 2017. “Anaerobic digestion of wastewater: effects of inoculants and nu-
trient management on biomethane rroduction and treatment.” All Graduate Theses and
Dissertations. 5699. https://digitalcommons.usu.edu/etd/5699

Schwede, S., Rehman, Z.U., Gerber, M., Theiss, C., Span, R., 2013. Effects of thermal
pretreatment on anaerobic digestion of Nannochloropsis salina biomass. Biores. Technol.
143, 505-511.

Sievers, D.M., and Brune, D.E. 1978. Carbon/nitrogen ratio and anaerobic digestion of swine
waste. Transactions of the ASAE 21 (3): 537-541

Soboh, Y.M., Sorensen, D.L.;,Sims, R.C. 2016. Upflow Anaerobic Sludge Blanket Reactor
Co-digestion of algae and acetate to produce methane. Water Environ. Res. 88, 2094–
2103.

Wang, X., Lu, C., Li, F., Yang, G. 2014. Effects of temperature and Carbon-Nitrogen (C/N)

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Corresponding Author: Ronald Sims, ron.sims@usu.edu

Citation: Peterson, J., Sims, R. (2021). Green energy through synergistic integration of wastewater and algae.
Academia Letters, Article 3732. https://doi.org/10.20935/AL3732.

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 ratio on the performance of anaerobic co-digestion of dairy manure, chicken manure, and
rice straw: focusing on ammonia inhibition. PloS one 9 (5), e97265.

Ward, A.J., Lewis, D.M., Green, F.B. 2014. Anaerobic digestion of algae biomass: a review.
Algal Res., 5, 204-214.

Yen, H.W., and Druce, D.E. 2007. Anaerobic co-digestion of algal sludge and wastewater to
produce methane. Biores. Technol. 98 (1): 130-134.

Academia Letters, October 2021 ©2021 by the authors — Open Access — Distributed under CC BY 4.0

Corresponding Author: Ronald Sims, ron.sims@usu.edu

Citation: Peterson, J., Sims, R. (2021). Green energy through synergistic integration of wastewater and algae.
Academia Letters, Article 3732. https://doi.org/10.20935/AL3732.