Seminar Presentation in the research article entitled:

Biogas productivity by co-digesting Taihu blue algae with corn

straw as an external carbon source

Article Detail:
Authors - Weizhang Zhong, Zhongzhi Zhang ⇑, Yijing Luo, Wei Qiao,
Meng Xiao, Min Zhang
Name of Journal- Bioresource Technology,
Volume - 114 (2012),
Period of Publication –December 2014,
Pages- 281–286
Web Link: https://doi.org/10.1016/j.biortech.2012.02.111

Suvranil Banerjee
ROLL NO: 002230601001
Academic Session : 2022-2023
Semester :1st
Master of Technology – Energy Science and Technology
School of Energy Studies
Jadavpur University
5th December, 2022
 Introduction 2

• The bloom of cyanobacteria is a ubiquitous phenomenon in eutrophic lakes, reservoirs, and polluted water in
many countries of the world.
• Over the past few years, several lakes in China, such as Chaohu Lake, Taihu Lake, and Dianchi Lake, among
others, have experienced increasingly massive cyanobacterial blooms (Qin, 2009).
• The excessive growth of cyanobacteria in lakes damages the natural functions of the lakes and threatens the
safety of the drinking water supply.
• For example, Taihu Lake, the third largest lake in China and which provides drinking water for more than 2
million people, caused serious toxicity problems because on an algal bloom during the summer of 2007 (Guo,
2007).
• The treatment of algal bloom in large lakes is a worldwide problem.
• Anaerobic digestion, coupled with energy production in the form of biogas and waste treatment, has been
employed for the treatment of several different kinds of organic wastes.
 Anaerobic Digestion and Algal Waste
3

• Anaerobic digestion is a spontaneous process, wherein anaerobic microorganisms decompose organic matter
and produce biogas primarily containing methane and carbon dioxide.
• The process does not require advanced dewatering or further chemical extraction.
• It not only reduces organic pollution but also provides a new source of energy.
• Several hundred thousand tons of algal waste may be skimmed annually from Taihu Lake alone, and such
large amounts of algae could be a potential substrate for biogas production.
• Golueke et al. (1957) published the first study on anaerobic digestion of microalgae (Scenedesmus spp. and
Chlorella spp.).
• Since then, a large number of research projects have been carried out. Early research efforts peaked in the late
1970s and early 1980s because of the first oil crisis.
• Species under investigation include several macroalgae such as Laminaria spp., Macrocystis spp. (Chynoweth
et al., 1993), marine alga Gracilaria ceae (Wise et al., 1979), and green marine alga Ulva spp. (Bruhn et al.,
2011).
• It can be concluded that algae are good feedstocks for anaerobic digestion, because of its high conversion rates
and efficiencies.
 Anaerobic Digestion and Algal Waste 4

• Except for biodegradation-resistant types, however, the low C/N ratio of algal sludge is also a serious problem to
anaerobic digestion.
• The optimum C/N range in feedstock for anaerobic digestion remains highly debated, although 20/1 to 30/1 is a
most acceptable range. The C/N ratio in the microalgal biomass is too low for anaerobic digestion (<10).
• Low C/N ratio feedstock could result in high total ammonia nitrogen (TAN) released and high volatile fatty acids
(VFAs) accumulated in the digester.
• The TAN and VFAs are both important intermediates and potential inhibitors in the anaerobic digestion process.
• One method to avoid excessive ammonia accumulation is to adjust low feedstock C/N ratios by adding high
carbon-content materials to improve digestion performance.
• Co-digestion of biomass waste with low C/N ratio, like algal sludge, with crop residues of high C/N ratio is
gaining momentum in many parts of the world.
• Crop residues, such as corn straw and wheat straw, are produced in large quantities in both China and other
countries every year, which, due to its organic nature, can be a valuable alternative feedstock for biogas
production (Li et al.,2009b).
 Anaerobic Digestion and Algal Waste 5

• The algal sludge used in the study was the mixtures of algae bloom and lake water. The mixtures were collected
from Meiliang Bay, Taihu Lake (120300N, 31270E). The Microcystis spp. was the dominant species in the
mixtures (>99%). The mixtures were stored at 4 oC before further use.
• The corn straw used in the study was collected from Nanshao Town, Changping District, Beijing City, China. The
straw was air-dried to final moisture contents of 7–8%, and then chopped with a paper chopper (PC500, Staida
Co., Tianjing, China) to a particle size of approximately 3 cm and then ground into 5–10 mm particles by a
hammer mill (FE130, Staida Co., Tianjing, China).
• The samples were then packed in plastic bags and placed under 62 ± 1% relative humidity and 2 ± 0.5 C until use.
• Taihu blue algae was rich in organics (volatile solids, VS), in which protein (nitrogen) content was as high as
61.76% of the total solids (TS).
• However, the lipid content of Taihu blue algae in the current study was 4.12% of the TS, which is relatively lower
than that of other species of microalgae that have lipid contents ranging from 20% to 50% of the TS of the
biomass (Singh and Gu, 2010).
 Methodology – Raw Material Source 6

• The algal sludge used in the study was the mixtures of algae bloom and lake water. The mixtures were collected
from Meiliang Bay, Taihu Lake (120300N, 31270E). The Microcystis spp. was the dominant species in the
mixtures (>99%). The mixtures were stored at 4 oC before further use.
• The corn straw used in the study was collected from Nanshao Town, Changping District, Beijing City, China. The
straw was air-dried to final moisture contents of 7–8%, and then chopped with a paper chopper (PC500, Staida
Co., Tianjing, China) to a particle size of approximately 3 cm and then ground into 5–10 mm particles by a
hammer mill (FE130, Staida Co., Tianjing, China).
• The samples were then packed in plastic bags and placed under 62 ± 1% relative humidity and 2 ± 0.5 C until use.
• Taihu blue algae was rich in organics (volatile solids, VS), in which protein (nitrogen) content was as high as
61.76% of the total solids (TS).
• However, the lipid content of Taihu blue algae in the current study was 4.12% of the TS, which is relatively lower
than that of other species of microalgae that have lipid contents ranging from 20% to 50% of the TS of the
biomass (Singh and Gu, 2010).
 Methodology – Inoculation 7

• The anaerobic sludge taken from an anaerobic digester with 4 L working volume, which had been cultivated and
used in a previous co-digestion process of algal sludge and corn straw at 35 oC, with 10 days retention time, was
used as inoculum.
 Methodology – Batch laboratory anaerobic digestion tests 8

• An anaerobic digestion test was performed in 150 mL serum bottles. First, the serum bottles were filled with 2 g
total substrates in VS with different proportions of algae and corn straw in the loading rate of 20 g VS L1.
• Digester 1 (D1) and Digester 2 (D2), were operated with corn straw only and algae only at a C/N ratios of 71/1
and 6/1, respectively and Digester 3 (D3), Digester 4 (D4), and Digester 5 (D5) were operated with the mixtures
of corn straw and algae at a C/N ratios of 16/1, 20/1, and 25/1, respectively.
• For batch digestion, the inoculum/substrate ratio (ISR) was 1.0. According to the previous research results by
Zeng et al. (2010), an ISR of 1.0 yields the maximum conversion rate of anaerobic digestion of Taihu blue algae.
The bottles were then seeded with 2 g anaerobic sludge in VS (about 34 mL on wet base).
• Distilled water was added up to a working volume of 100 mL.
 Methodology – Batch laboratory anaerobic digestion tests 9

• The pH was measured, and the bottles were gassed with N2 before 2–3 drops of sodium sulfide were added to
ensure anaerobic conditions.
• The samples were immediately sealed using rubber septa and aluminum crimp caps with drilled holes. A slender
glass tube with a silicone tube was inserted to export gas for biogas volume and composition measurement.
• Once sealed, the bottles were placed in an incubator and shaken at a constant mesophilic temperature of 35 oC and
at a speed of 150 rpm.
• Daily biogas production was recorded by quantifying water displacement.
• For the determination of endogenous methane production, blanks containing only anaerobic sludge were run. The
incubation time was approximately 30 days.
• All the experiments were carried out in triplicate and the results were expressed as means.
 Methodology – Batch laboratory anaerobic digestion tests 10

• The pH was measured, and the bottles were gassed with N2 before 2–3 drops of sodium sulfide were added to
ensure anaerobic conditions.
• The samples were immediately sealed using rubber septa and aluminum crimp caps with drilled holes. A slender
glass tube with a silicone tube was inserted to export gas for biogas volume and composition measurement.
• Once sealed, the bottles were placed in an incubator and shaken at a constant mesophilic temperature of 35 oC and
at a speed of 150 rpm.
• Daily biogas production was recorded by quantifying water displacement.
• For the determination of endogenous methane production, blanks containing only anaerobic sludge were run. The
incubation time was approximately 30 days.
• All the experiments were carried out in triplicate and the results were expressed as means.
 Methodology – Analytical Methods 11

• Biogas volume was monitored every day through the water displacement method and the corresponding
cumulative biogas volume calculated. The measured volume was then converted to a volume of gas at
standard temperature and pressure using the ideal gas law.
• CH4 and CO2 were analyzed using a gas chromatograph (Auto System, Shimadzu, Japan) equipped with a
packed column (Shimadzu, 6‘ ×1.8“ OD, 80/100, Japan) and a thermal conductivity detector (TCD).
• The temperatures of the oven, injector port, and TCD were 120, 150, and 150 C, respectively.
• The carrier was hydrogen-operated at a flow rate of 30 Ml/min at 60 C.
• A standard gas (Zhaoge Co., Beijing, China) composition of 5.0% N 2, 60.1% CH4, and 34.9% CO2 was
used for the calibration of the system.
• CH4 production was determined by multiplying the CH 4 content with the amount of biogas production.

Gas analyses were carried out two or three times a week in the following weeks. A 250 µL pressure-tight
gas syringe (VICI, Precision Sampling Inc., USA) was used for gas sampling.
 Methodology – Analytical Methods
12

• The TS and VS of the algae sludge, corn straw, anaerobic sludge and their mixture, the total carbon (TC),
and total nitrogen (TN) were measured in triplicate according to the APHA standard methods.
• The pH value was directly measured from liquid samples with a digital pH meter (CHN868, Thermo
Orion, America).
• Lipids were determined by a Soxhlet system at 65 C with more than 60 circulations using methylene
dichloride as an extractive reagent. The sample weights before and after extraction were used to calculate
the lipids content.
• Protein was calculated by multiplying TN by 6.25. Carbohydrate was calculated as the fraction of VS
remaining after the subtraction of protein and lipids (Li et al., 2009a).
• Liquid analyses of fermentation liquor, including total volatile fatty acids (TVFA) and ammonia, were
conducted in triplicate before and after anaerobic digestion tests. Soluble fractions were obtained after
centrifugation (SIGMA, 10,000 rpm for 30 min at 2 C).
• TVFA content was monitored using a gas chromatograph (Auto System, Shimadzu, Japan) equipped with
a fused silica capillary column (Shimadzu, 15 m 0.53 mm 0.5 lm film thickness) and a flame ionization
detector (200 C) with hydrogen as the carrier gas.
Results and Discussion
13
Results and Discussion
14
Results and Discussion
15
Results and Discussion 16
Results and Discussion 17

• For the same amount of substrate of 2 g VS loaded, it was found that the co-digestions with
mixture of algae sludge and corn straw achieved 16.52–51.60% more biogas yield and 18.95–
62.09% more methane yield, than that of the single-digestions with algae sludge only,
respectively .
• The differences were significant (p < 0.05). This means that based on the same amount of algae
sludge and corn straw feedstock, more bioenergy can be generated when the co-digestion
process is applied. This increase was attributed to synergistic effects from the co-digestion of
algae sludge and corn straw.
• One possible explanation for this is that algal biomass not only plays a role as a nitrogen source
supplier in this co-digestion process, but also supplies nutrients to the digester microbes after
the degradation of algal biomass.
• Co-digestion of algal sludge and corn straw could efficiently balance feedstock carbon and
nitrogen. In turn, an optimal feedstock C/N ratio was likely beneficial to methanogen activity
and resulted in decreased VFA concentrations due to higher conversion rate to methane.
Conclusion 18

• A batch anaerobic test was conducted to evaluate the effects of adding high carbon content of corn straw to the
digestion of Taihu blue algae to attain an optimal C/N ratio for higher methane yield. The addition of corn
straw in algae at a C/N ratio of 20/1 increased methane yield by 61.69 % at 325 mL g -1 VS-1 (compared with
201 mL g1 VS1 of algae digestion alone), followed by C/N ratios of 16/1 and 25/1, all operated at 20 g VS L1
and 35 C. The results suggest the optimal C/N ratio for co-digestion of algae with corn straw is 20/1. The
findings could offer options for efficient methane production and waste treatment.
• The results from this study clearly indicate that significant increases in volumetric methane production can be
achieved by adding carbon-rich corn straw to the co-digestion process with Taihu blue algae. Among the three
C/N ratios tested, the C/N ratio of 20/1 was found to be the best in terms of methane productivity, which
increased by 61.69% in the current study, as compared to algae digestion alone. The results also showed that
co-digestion of Taihu blue algae with corn straw could be one of the options for efficient methane production
and waste treatment.
Reference 19

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THANK YOU
Suvranil Banerjee​
suvranil.banerjee@gmail.com