See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/348450626

Effect of co-digestion of tylosin fermentation dreg and food waste on

anaerobic digestion performance

Article  in  Bioresource Technology · January 2021

DOI: 10.1016/j.biortech.2021.124693

CITATIONS READS

13 49

6 authors, including:

Ming Gao Xinxin Ma

University of Science and Technology Beijing University of Science and Technology Beijing
66 PUBLICATIONS   794 CITATIONS    9 PUBLICATIONS   49 CITATIONS   

SEE PROFILE SEE PROFILE

Dong Xie
University of Science and Technology Beijing
4 PUBLICATIONS   18 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Fly ash View project

Optimization of VFAs production from Food Waste by Two Stage Fermentation View project

All content following this page was uploaded by Ming Gao on 28 January 2021.

The user has requested enhancement of the downloaded file.

 Bioresource Technology 325 (2021) 124693

Contents lists available at ScienceDirect

Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Effect of co-digestion of tylosin fermentation dreg and food waste on

anaerobic digestion performance
Ming Gao a, b, 1, Min Yang a, 1, Xinxin Ma a, Dong Xie a, Chuanfu Wu a, b, *, Qunhui Wang a, b, c
a
School of Energy and Environmental Engineering, University of Science and Technology Beijing, Beijing 100083, PR China
b
Beijing Key Laboratory on Disposal and Resource Recovery of Industry Typical Pollutants, University of Science and Technology Beijing, Beijing 100083, PR China
c
Tianjin College, University of Science and Technology Beijing, Tianjin 301830, PR China

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• This study investigated the co-digestion

of TFD and food waste.
• pH, TVFA and acetic acid were the most
important factors affecting CH4
production.
• Tylosin concentration decreased signifi­
cantly after anaerobic digestion.
• This study provided a feasible method
for the treatment of TFD.

A R T I C L E I N F O A B S T R A C T

Keywords: In this study, the effects of adding different food waste proportions (volatile solids ratio of 3:7, 5:5, 7:3) to tylosin
Tylosin fermentation dreg on anaerobic digestion were investigated. The results showed that the co-digestion group
Antibiotic fermentation dregs (294–399 mL⋅g-VS− 1) increased methane production by 14.8%–55.5% compared with tylosin fermentation dreg
Anaerobic co-digestion
alone-digestion (256 mL⋅g-VS− 1). The correlation analysis showed that pH, total volatile fatty acids and acetic
Correlation
Synergy
acid were the most important factors affecting cumulative methane production. Tylosin in the solid and liquid
phases decreased significantly after anaerobic digestion, indicating that tylosin could be effectively removed by
co-digestion, and the total removal rate was 68.2%–83.7%. Therefore, due to the satisfactory methane yield and
the tolerable tylosin removal rate, it is feasible to make the co-digestion of tylosin fermentation dreg and food
waste.

1. Introduction development of livestock and disease prevention (Yin et al., 2016).

However, when tylosin is produced by fermentation, a large number of
Tylosin is a macrolide antibiotic that is often used in the growth and fermentation dregs is generated during the production process, and the

* Corresponding author at: Department of Environmental Engineering, School of Energy and Environmental Engineering, University of Science and Technology
Beijing, 30 Xueyuan Road, Haidian District, Beijing 100083, PR China.
E-mail address: wucf@ustb.edu.cn (C. Wu).
1
Ming Gao and Min Yang contributed equally to this work.

https://doi.org/10.1016/j.biortech.2021.124693
Received 12 November 2020; Received in revised form 3 January 2021; Accepted 6 January 2021
Available online 11 January 2021
0960-8524/© 2021 Elsevier Ltd. All rights reserved.
M. Gao et al. Bioresource Technology 325 (2021) 124693

yield ratio of the two is about 1:8–1:12. The direct discharge of anti­ Table 1
biotic fermentation dregs causes the spread of residual antibiotics in the The specific properties of the materials.
soil, surface water, groundwater, and other environments, seriously Properties Tylosin fermentation dreg Food waste Sludge
endangering the environment and the health of the food chain (Chen a
TS (%) 32.6 26.0 8.81
et al., 2017). In view of this, antibiotic fermentation dregs are currently VS(%)a 25.9 24.1 5.12
included in the “National Hazardous Waste List” (Zhang et al., 2018) and C(%)b 41.3 48.6 30.0
need to be disposed of as hazardous waste. The safe and reasonable N(%)b 4.75 1.97 2.40
disposal of antibiotic fermentation dregs has become a bottleneck H(%)b 6.50 7.13 –
O(%)b 46.9 40.2
problem restricting the development of the pharmaceutical industry.
–
C/N 8.69 24.3 12.5
Antibiotic fermentation dregs contain a large amount of biodegradable Tylosin (mg/g) b
8.90 – –
organic matter, such as proteins, amino acids, and polysaccharides (Cai pH 6.02 5.11 7.82
et al., 2017). Using these fermentation dregs to produce biogas through a
refers to a wet basis. b refers to a dry basis.
anaerobic digestion technology can reduce the amount of resource
wasted and environmental pollution caused by improper treatment. It
will also improve the energy supply structure and sanitation conditions Table 2
as well as increasing the added value of these fermentation dregs (Zhang The basic parameters of each experimental group.
et al., 2015). Some researchers have used penicillin fermentation dregs Reactor Tylosin Food Sludge Tylosinconcentration
and cephalosporin antibiotic fermentation dregs for anaerobic digestion numbers fermentation dreg waste (g) a (mg/L) a
(Li et al., 2015; Zhong et al., 2015), and proved the feasibility of this (g)a (g) a
treatment technology. However, using different types of antibiotic AD10:0 8.22 0 41.8 480
fermentation dregs, the methane production yield and the degradation AD7:3 5.74 2.64 41.6 335
rates of antibiotics anaerobic digestion process are significantly different AD5:5 4.09 4.39 41.5 238
AD3:7 2.45 6.14 41.4 143
(Ni et al., 2020). To date, no studies have been carried out on the
AD0:10 0 8.78 41.5 0
anaerobic digestion of tylosin fermentation dreg (TFD). Residual tylosin
a
in the TFD is an important factor that affects its resource utilization. Refers to a wet basis.
Researchers have explored the effects of adding tylosin in livestock
manure on the anaerobic digestion process. Mitchell et al. (2013) found and various parameters in the anaerobic digestion process, and the
that when tylosin concentration was less than 91 mg⋅L− 1, it had no in­ removal of tylosin in the anaerobic digestion process was investigated.
hibition on methane production; Yin et al. (2019) found that when the This study could provide a strong scientific basis for the energy utili­
initial concentration of tylosin was less than 25 mg⋅kg-TS− 1, it can be zation of TFDs.
completely removed by anaerobic digestion; Stone et al. (2009) and
Zhang et al. (2019) found that tylosin increased the abundance of ace­ 2. Materials and methods
ticlastic methanogens and promoted the aceticlastic methanogenic
pathway. But the effect of tylosin in TFD has not been studied yet. 2.1. Substrate and inoculum
Considering that many refractory substances in antibiotic fermentation
dregs and the low efficiency of direct anaerobic digestion, researchers The TFD used in this experiment was from Shandong, China. Food
have also studied pretreatment methods, such as hydrothermal treat­ waste was collected from a student canteen at the University of Science
ment (Li et al., 2015), alkali treatment (Zhong et al., 2015), and alkali and Technology Beijing. After removing the bones, plastic, and paper
heating (Li et al., 2016a, 2016b), to destroy the cell wall and increase the products, the remaining waste was drained and sliced, and then stored in
dissolution rate of intracellular substances, and then improve the utili­ a low-temperature refrigerator at − 20 ◦ C. Anaerobic sludge was
zation rate of hydrolytic bacteria. Although these pretreatment methods collected from a rural biogas station in the Shunyi District, Beijing,
can significantly improve methane production, they also increase the China. The anaerobic sludge was pre-incubated at 37 ℃ for two (2)
energy and chemical input. weeks before being used. After pre-incubation, glucose was selected as
Co-digestion is an effective method to improve the methane pro­ the standard product for sludge activity test, and its methane yield
duction efficiency. It can establish a positive complementary balance reached 387 mL⋅g-VS− 1, which proved that the sludge activity was good.
between materials and play a role in diluting toxic substances and The chemical characteristics of the substrate and sludge are shown in
providing nutrients. The C/N ratio of food waste is high, and the Table 1.
degradable substances are rich, which is conducive to the growth and
reproduction of microorganisms at the early stage of the anaerobic 2.2. Batch anaerobic co-digestion experiment
digestion process (Ren et al., 2018; Zhang et al., 2014). Using food waste
as a co-substance to achieve co-digestion with TFD can solve the prob­ The batch experiment was carried out with 100 mL anaerobic
lem of insufficient microbial substrate caused by the slow degradation fermentation bottles as the reaction vessel, and their work volume was
rate of TFD. On the other hand, it can also adjust the C/N ratio to pro­ 50 mL, which was convenient for observing the change in the material
mote the degradation of antibiotic fermentation dregs itself. Further­ state. The substrate to inoculum ratio (SIR) was 1:1 (VS ratio) to
more, there is no need for co-digestion to build a new reactor for each maintain successful operation based on our previous research (Wu et al.,
substrate, and different types of substrates can use the same reactor, 2015), and the reactors were flushed with nitrogen for 1 min to produce
thereby reducing the reactor construction cost to a certain extent anaerobic conditions. After sealing, the bottles were placed in an air
(Chuenchart et al., 2020; Ma et al., 2019). bath oscillator for batch anaerobic fermentation (37 ◦ C, 120 rpm). Five
This study evaluates the feasibility of TFDs and food waste under experimental groups were set up, and their VS ratios of TFD and food
different volatile solid (VS) mixing ratios (10:0, 7:3, 5:5, 3:7 and 0:10) waste were 10:0, 7:3, 5:5, 3:7, and 0:10, respectively. The five experi­
under mesophilic anaerobic digestion conditions. The effects of co- mental groups were recorded as AD10:0, AD7:3, AD5:5, AD3:7, and
digestion on cumulative methane production, biodegradability, AD0:10. In addition, a blank group with only sludge fermentation was
methane production rate, degradation rate and the process performance set up. All the data in this study were obtained after deducting the
were studied. The digestion process was modelled and kinetics analysis control group. The basic information on the substrate and inoculum in
were carried out. Correlation analysis and principal component analysis each experimental group is shown in Table 2. Ten parallel experiments
(PCA) were used to study the relationship between methane production were carried out in each group, and the methane production of each

2
M. Gao et al. Bioresource Technology 325 (2021) 124693

Fig. 1. Cumulative methane production and biodegradability under different substrate mixing ratios. TFD: tylosin fermentation dreg; FW: food waste; CMY: cu­
mulative methane yield (mL/g-VS); TMY: theoretical methane yield (mL/g-VS) as calculated by Eqs. (1)–(3).

reactor was measured at intervals, and the average value was taken as
(4a + b − 2c − 3) 22.4 × 1000
the experimental result. Then, one of the reactors in each group was TMY(mL/gVS) = × (2)
8 12a + b + 16c + 14
opened by a destructive sampling method to test pH, ammonia nitrogen,
VFA, alkalinity, and other parameters. The sampling time was 0, 1, 4, 8, CMY
15, 21, 25, 28, 32, 40, 69 days. Biodegradability(%) = × 100% (3)
TMY
Note: CMY refers to the cumulative methane yield (mL⋅g-VS− 1), and
2.3. Analysis method TMY refers to the theoretical methane yield (mL⋅g-VS− 1).
In this study, three commonly used models: the modified Gomepertz
2.3.1. Routine analysis model (Eq. (4)), the first-order kinetic model (Eq. (5)), and cone model
The methane volume was measured using the alkali solution (3 (Eq. (6)) were selected to simulate the cumulative methane production
mol⋅L− 1 NaOH) drainage method. Pass the produced biogas into an of different experimental groups (Yu et al., 2018a).
inverted measuring cylinder filled with alkali solution, stand still for 1 { }
(RM × e)
min, during which CO2 was completely absorbed, and then read the M(t) = fd × exp − exp[ (λ − t) + 1] (4)
fd
reduced volume of alkali solution that is also the volume of methane
(ignoring a very small amount of hydrogen). The total solid and volatile
M(t) = [1 − exp( − kh × t)] (5)
solid contents were tested according to the State Environmental Pro­
tection Administration (2002). The pH value was measured by pHS-3C fd
type digital acidity (INESA Scientific Instrument Co., Ltd, Shanghai, M(t) = n (6)
1 + (kh × t)−
China). The alkalinity was determined by bromocresol green methyl red
indicator titration. Total ammonia nitrogen was measured by ultra­ where M(t) is the cumulative methane production at a given time (mL⋅g-
violet–visible spectrophotometry (HJ 535-2009). Total volatile fatty VS− 1), fd is the final methane production (mL⋅g-VS− 1), kh is the hydro­
acids (TVFAs) includes acetic, propionic, butyric, iso-butyric, valeric, lysis rate constant (d− 1); RM is the maximum methane production rate
and isovaleric acids. The measurement methods of ethanol and VFA was (mL⋅g-VS− 1⋅d− 1), λ is the duration of lag phase (d), t is the time (d), e is
described by Ma et al. (2020). Firstly, samples were centrifuged for 10 the exponential constant, taking 2.7183, and n is the shape factor.
min at 12,000 r⋅min− 1. Secondly, the supernatant was diluted and In accordance with the co-substrate ratios of the substrates and the
filtered with 0.2 µm microporous membrane. The samples were then biogas yields obtained during the separate digestion, the estimated
determined on Shimadzu CP3800 gas chromatography equipped with a biogas yields for each experimental group were obtained using Eq. (7)
DB-FFAP capillary column (30 m × 0.53 mm × 0.5 µm, Agilent Tech­ (Zhen et al. 2016).
nologies, Inc., Beijing, China) and a flame ionization detector. The
tylosin concentration was determined by HPLC after purification and Bestimated(t) = Bmeasured− FW(t) × X1 + Bmeasured− TFD(t) × X2 (7)
extraction and the detection limit was 1 mg⋅kg− 1. Statgraphics 18 soft­
ware was used for the correlation analysis, R language software was where Bestimated(t) (mL⋅g-VS− 1) is the estimated biogas yield at a given
used for the PCA, and origin 2018 was used for data processing and time, Bmeasured− FW(t) (mL⋅g-VS− 1) is the measured biogas yield of the FW
mapping of other data. alone, Bmeasured− TFD(t) (mL⋅g-VS− 1) is the measured biogas yield of the
TFD alone (mL⋅g-VS− 1), X1 (%) is the percentage of the FW in the co-
2.3.2. Biodegradability, kinetic and synergy substrates, X2 (%) is the percentage of the TFD in the co-substrates
According to the method of Ma et al. (2019), the biodegradability and t (day) is the digestion time.
was calculated as follows: The synergies from the co-digestion of FW and TFD were calculated
using Eq. (8).
4a− b− 2c+3 4a+b− 2c− 3 4a− b+2c+3
Ca Hb Oc N+ H2 O→ CH4 + CO2 +NH3 ∑
4 8 8 Bmeasured(t)
Synergy = ∑ × 100% (8)
(1) Bestimated(t)

3
M. Gao et al.
Fig. 2. The daily methane yield at different mixing ratios and its box diagram. TFD: tylosin fermentation dreg; FW: food waste.
If synergy > 100%, then co-digestion has a positive synergistic effect.
If synergy < 100%, then co-digestion has a negative synergistic effect. If
synergy = 100%, then no synergistic effect exists (Kim et al., 2019).
3. Results and discussion
3.1. Methanogenic performance
3.1.1. Cumulative methane production and biodegradability
Methane production is the key parameter for evaluating the perfor­
mance of an anaerobic reactor. Fig. 1 shows the cumulative methane
production and biodegradability of the co-digestion with TFD and FW at
different mixing ratios. As shown in Fig. 1 (a), the cumulative methane
production in different groups is significantly different (p > 0.05). The
highest was the FW digested alone (487 mL⋅g-VS− 1), while the lowest
was the TFD digested alone (256 mL⋅g-VS− 1). In the co-digestion group
(294–399 mL⋅g-VS− 1), the higher the mixing ratio of FW, the higher the
cumulative methane production. The cumulative methane production of
the five groups ranked as AD0:10 > AD3:7 > AD5:5 > AD7:3 > AD10:0
(TFD:FW). The cumulative methane production of the co-digestion
groups was 14.8–55.5% higher than that of single TFD digestion.
Therefore, adding food waste to the TFD digestion process can signifi­
cantly improve the methane production performance of the anaerobic
system. However, it is worth noting that this kind of improvement was
mainly due to the higher methane potential of food waste than the TFD.
On the contrary, the synergy of the co-digestion group was only 86.9%-
95.5% according to the Eqs. (7)-(8), indicating that the synergy of TFD
and food waste was poor. The reasons may include: (1) The previous
research showed that when the sludge inoculation ratio during the
anaerobic fermentation of food waste was approximately 1, the anaer­
obic fermentation performance of food waste was stable, the fermenta­
tion system didn’t produce acidification, and the methane production
reaches the theoretical level (Wu et al., 2015). Thus, the co-digestion of
TFD and food waste was difficult to further increase the methane po­
tential of any single substrate. (2) Due to the low C/N and high tylosin
concentration of TFD, it reduced the methane production potential of
food waste to a certain extent. (3) Although the addition of food waste
may increase the methane production potential of TFD, the increase was
less than the inhibition of TFD on the methane production potential of
food waste. According to Table 1 and Eqs. (1)–(3), the theoretical
methane production and biodegradability of different substrates can be
calculated. As shown in Fig. 1(b), the biodegradability of the five
experimental groups reached more than 79.4%, which indicated that the
substrates of the five experimental groups were basically consumed, and
4
Bioresource Technology 325 (2021) 124693
this was a successful anaerobic digestion experiment (Wu et al., 2018).
The theoretical methane production of TFD was 301 mL⋅g-VS− 1, which
was significantly lower than that of FW (513 mL⋅g-VS− 1), but its
biodegradability was good, reaching 85.1%, indicating that TFD is a
good substrate for anaerobic digestion. The cumulative methane pro­
duction and theoretical methane production increased with the increase
in the proportion of FW added, but the biodegradability fluctuated only
slightly (the fluctuation range was − 5.70–3.70%), indicating that co-
digestion had little effect on biodegradability.
Fig. 1 Cumulative methane production and biodegradability under
different substrate mixing ratios. TFD: tylosin fermentation dreg; FW:
food waste; CMY: cumulative methane yield (mL⋅g-VS− 1); TMY: theo­
retical methane yield (mL⋅g-VS− 1) as calculated by Eqs. (1)–(3).
3.1.2. Methane production rate
Fig. 2 shows the daily methane production rate at different mixing
ratios. The daily methane production rate can reflect the peak situation
of methane production, and different methane production peaks can
reflect the difference in anaerobic fermentation kinetics (Zhen et al.,
2016). The emergence of the methane production peak was due to the
presence of many components that can be easily used by microorgan­
isms in the reactor. The first gas production peak may be due to the pre-
existing soluble components in the raw materials, which can be quickly
converted into methane. With the decrease in substrates that metha­
nogens can directly use, macromolecular substances in raw materials
will be gradually degraded by hydrolysis bacteria to produce new sol­
uble components, and then produce new gas production peaks (Zhen
et al., 2016). However, previous studies only qualitatively determined
the number of gas production peaks from the change in the daily gas
production rate, which has a certain degree of inaccuracy.
Fig. 2 The daily methane yield at different mixing ratios and its box
diagram. TFD: tylosin fermentation dreg; FW: food waste.
This study found that the abnormal values of daily gas production
rate of different experimental groups were obtained from Fig. 2 (b) (i.e.,
the values beyond the upper limit of the box plot and have been circled
by a gray ellipse), then, the occurrence time of these abnormal values
was determined from Fig. 2 (a), and the number of gas production peaks
was determined according to the occurrence time distribution. This
method can accurately summarize the number and occurrence time of
the gas production peak in different reactors. In this study, the highest
gas production peak appeared in the five experimental groups on the
first day, and the second gas production peak appeared at the 26th, 25th,
24th, and 15th days of AD7:3, AD5:5, AD3:7, and AD0:10 groups,
respectively, while the 10:0 group maintained stable gas production
M. Gao et al. Bioresource Technology 325 (2021) 124693

Table 3 (Yu et al., 2018b). In this study, the R2 values of these three models were
Kinetic paraments using three different models to simulate the methane pro­ generally above 0.97, which indicated that these models fit accurately,
duction of different mixing ratio groups. and the results are reliable (Yu et al., 2018b). The hydrolysis rate con­
Paraments AD10:0 AD7:3 AD5:5 AD3:7 AD0:10 R2 stant (Kh) is an indicator of biodegradability and fermentation efficiency
kh a
0.014 0.018 0.025 0.027 0.060 0.978 ~ 0.993
of substrates. The higher the value, the faster the degradation rate of
kh b 0.019 0.023 0.036 0.043 0.082 0.986 ~ 0.995 organic matter and the methane production rate (Koch et al., 2015). Kh
λc (d) 0.874 0.229 0.654 2.05 1.38 0.994 ~ 0.998 values from the first-order kinetic model and cone model were consis­
Rm c(mL/d) 5.26 6.36 8.06 11.1 22.2 0.994 ~ 0.998 tent in this study. The Kh value of the TFD single digestion was only
a: the first-order kinetic mode; b: the cone mode; c: the modified Gomepertz 23.3% of FW digested alone, and all the Kh values of the co-digestion
model. group were improved. However, due to the limitation of the TFD, the
maximum value was only about half of the single digestion of FW, which
from 10 to 45 days without any gas production peak. The components in indicated that the co-digestion of FW and TFD had no significant positive
the FW are easy to hydrolyze, so when it is digested alone, the second gas effect on the hydrolysis rate constant. Lag stage (λ) is an inherent bio­
production peak appears at the earliest. However, the mycelium, re­ logical phenomenon that reflects the delayed response of organisms to
sidual medium, metabolic intermediates, and organic solvents contained environmental changes (Li et al., 2016b), which can be obtained using
in TFD were difficult to use for hydrolytic bacteria, and it was difficult to the modified Gomepertz model. It can be found that the extreme values
form a large number of soluble components in a short time (Cai et al., of this parameter appeared in the co-digestion group, the minimum
2017). Therefore, it was difficult to form a second gas production peak; value appeared in the AD7:3 group (0.229 d), and the maximum value
thus, AD 0:10 maintains a long continuous and stable gas production. appeared in the AD3:7 group (2.05 d). The lag time of the FW alone-
When FW was introduced for co-digestion, the second gas production digestion was longer than that of the TFD alone, which meant that the
peak appeared, which indicated that co-digestion improved the disso­ mixed ratio of substrate may be an important factor affecting the lag
lution efficiency of organic matter and the biodegradability of co- time. Under the appropriate mixing ratio, co-digestion can significantly
substrates, and induced or accelerated the heterogeneous degradation shorten the lag period.
process.
3.2. System stability factors
3.1.3. Kinetic analysis
In this study, three common models were selected: the modified In the anaerobic digestion process, pH, TVFA, alkalinity, TVFA/
Gomepertz model (Eq. (4)), the first-order kinetic model (Eq. (5)), and alkalinity, total ammonia nitrogen (TAN), various organic acids and
cone model (Eq. (6)). By simulating the cumulative methane production ethanol have been proven to be important factors affecting system sta­
in Fig. 1 (a), the key indicators reflecting anaerobic digestion efficiency, bility and cumulative methane production (Ma et al., 2020). These
such as the hydrolysis rate of particulate organic matter, duration of lag factors can significantly affect the chemical environment of the system,
period, and maximum gas production rate, can be calculated (Table 3) thereby affecting the growth, metabolism, and methane production of
effective microorganisms (Ma et al., 2020; Ren et al., 2018).

Fig. 3. Changes of pH value, TVFA, alkalinity, TVFA/alkalinity in different digestion groups over time.

5
M. Gao et al.
Fig. 4. Correlation analysis of anaerobic digestion factors. CMY: cumulative methane yield; TVFA: total volatile fatty acid; TAN: total ammonia nitrogen.
Fig. 3 reflects the changes of pH, TVFA, alkalinity, TVFA/alkalinity
in different groups during digestion process. The sampling points were
0, 1, 4, 8, 15, 21, 25, 28, 32 , 40, 69 days. It can be seen from Fig. 3(a)
that the initial pH values of the five groups were all around 7.74, and all
dropped to the lowest point on the first day, Subsequently, the pH values
gradually recovered and almost all returned to the initial state on the
25th day, and finally rose to between 7.91 and 8.09. In the batch
anaerobic digestion process, the pH value almost always decreased first
and then increased, which was because the easily degradable compo­
nents in the substrate were rapidly utilized by the hydrolysis acidifica­
tion bacteria (Wu, et al., 2015). In this study, the higher the proportion
of the food waste, the greater the drop in the pH value on the first day,
because there were more pre-existing soluble components in the food
waste, which also led to the correspondingly highest daily methane
production rate on that day (Fig. 2 (a)). The change trend of TVFA (Fig. 3
(b)) was the opposite of pH, showing a trend of rising first and then
falling and the maximum appeared on the 4th day. The higher the
proportion of TFD, the higher the TVFA concentration at the highest
point, which may be because the residual tylosin in the TFD inhibited
the further utilization of organic acids by methanogens (Mitchell et al.,
2013). At the end of the fermentation, the TVFA concentration of each
group was similar, maintaining around 1.25 g⋅L− 1. The metabolism of
ammonia will produce bicarbonate alkalinity, and the protein content in
TFD and food waste were both high, causing the alkalinity of each group
gradually increased (Fig. 3(c)), which also showed that there was no
acidification inhibition in the entire anaerobic digestion process. TVFA/
6
Bioresource Technology 325 (2021) 124693
alkalinity is a commonly used evaluation index of system stability, and
usually affected by the SIR in the batch experiment. TVFA/alkalinity in
the batch experiment usually affected by the SIR. Choosing a suitable
SIR can not only maximize the processing efficiency of the reactor, but
also ensure its stable operation. Fig. 3(d) reflects that the TVFA/alka­
linity ratio of each group has no significant difference (p < 0.05), and it
increased first and then decreased. In the batch anaerobic digestion, the
suitable TVFA/alkalinity ratio has a pretty wide range and its threshold
is generally 0.8–1.0 in batch reactors (Wang et al., 2012). Although the
maximum value reached 1.65, that was a short-lived situation unique to
batch reactors and did not mean that the system was unstable.
Fig. 3 Changes of pH value, TVFA, alkalinity, TVFA/alkalinity in
different digestion groups over time
Fig. 4 shows the heat map of the Spearman correlation coefficient,
PCA diagram between cumulative methane production (CMY) and
various factors, and the relationship of CMY and ammonia nitrogen. In
Fig. 4 (a), it was found that pH, alkalinity, and propionic acid/acetic acid
were positively correlated with CMY, while various organic acids, TVFA,
and TVFA/alkalinity were negatively correlated with CMY. The corre­
lation coefficient between CMY and propionic acid/acetic acid was
greater than 0.9, and the correlation coefficient with valeric acid was
less than − 0.9. It is worth noting that TVFA/alkalinity, a widely vali­
dated index of system stability, had a correlation coefficient greater than
0.9, with acetic acid and butyric acid, indicating that acetic acid and
butyric acid were more sensitive to system changes. The correlation
coefficients of some organic acids were all greater than 0.9 (acetic and
M. Gao et al.
Fig. 5. Removal of tylosin in liquid and solid phases at different mixing ratios.
butyric acids, isovaleric and iso-butyric acids, butyric and valeric acids),
which was related to their transformation pathway in the anaerobic
digestion reaction. Then, the factors with strong correlation were
selectively retained, and the remaining factors were analyzed by PCA to
obtain Fig. 4 (b). Ma et al. (2020) can be used to interpret PCA diagrams.
The contribution rate of the two principal component factors (PCA1 +
PCA2) was 81.8%, which indicated that we have successfully identified
the principal components of the influencing factors.
The four factors of pH, acetic acid, TVFA, and alkalinity had the
strongest correlation with CMY, but the strong correlation of alkalinity
was due to the continuous increased as the anaerobic digestion process
proceeded, which indicated that pH, TVFA, and acetic acid were the
most important parameters that need to be focused and adjusted during
the co-digestion of TFD and food waste. In this study, the higher the pH
(7.02 ~ 8.16), or the lower the TVFA (1.00 ~ 3.78 g⋅L− 1L) and acetic
acid concentration (0.221 ~ 1.46 g⋅L− 1), the better the conversion of the
substrates into methane. Moreover, it is worth noting that ammonia
nitrogen is also an important factor affecting the stability of the anaer­
obic digestion system (Wu et al., 2018), especially for some substrates
with high protein content (i.e. low C/N). It can be seen from Fig. 4(c)
that the ammonia nitrogen concentrations at the end of digestion pro­
cess had a significant negative correlation with the CMY (R2 = 0.9505).
The result showed that the higher the proportion of TFD in the co-
substrates or the lower the C/N, the higher the ammonia nitrogen con­
centration in the digestate at the end of digestion, causing the inhibition
for the methane production performance to a certain extent (Rajagopal
et al., 2013). Furthermore, the TFD used in this study contains a certain
amount of tylosin and high protein content. During anaerobic digestion,
it will metabolize more protein and produce more ammonia nitrogen.
Therefore, antibiotic inhibition and ammonia nitrogen inhibition may
be two concerns for anaerobic digestion of antibiotic fermentation
residues.
Fig. 4 Correlation analysis of anaerobic digestion factors. CMY: cu­
mulative methane yield; TVFA: total volatile fatty acid; TAN: total
ammonia nitrogen.
7
Bioresource Technology 325 (2021) 124693
3.3. Removal of tylosin
Fig. 5 showed that the initial amount of tylosin of the four fermen­
tation groups containing TFD were 149–491 mg⋅L− 1, mostly in the solid
phase, and only about 20% in the liquid phase. The content of tylosin in
solid and liquid phase decreased obviously after anaerobic digestion, the
total removal rate of tylosin was 68.2%–83.7%, and the residual tylosin
was 47.2–83.6 mg⋅L− 1. The concentration of tylosin in the solid phase
decreased more, and the removal rate reached 79.4%–89.9%, possibly
because tylosin in the solid phase could be transferred to the liquid
phase by dissolution. The removal rate of tylosin in the liquid phase was
50.4%–58.4% except for the AD3:7 group, and the residual amount was
between and 22.9–40.1 mg⋅L− 1. The removal rate of tylosin in the
anaerobic digestion process is often related to its initial concentration.
When the initial concentration is low, the final residual tylosin is less
and the removal rate is higher (Mitchell et al., 2013; Yin et al., 2019). In
this study, TFD still remained a certain degree of tylosin after anaerobic
digestion treatment, which was related to its high initial concentration.
In conclusion, anaerobic digestion can remove tylosin in TFD to a certain
extent, even if the initial concentration was as high as 500 mg⋅L− 1, the
removal rate was still more than 68%. In the biological process,
biodegradation and hydrolysis are the main ways to remove antibiotics
(Wang et al., 2018). However, macrolide antibiotics are stable at
25–50 ◦ C and relatively neutral pH (6–8) (Paesen et al., 1995). In this
study, the temperature and pH of anaerobic fermentation were around
37 ◦ C and 7.5, respectively. Therefore, in the stable mesophilic anaer­
obic digestion process, the main removal method of tylosin is the
decomposition of anaerobic microorganisms.
Fig. 5 Removal of tylosin in liquid and solid phases with different
mixing ratios.
4. Conclusion
When adding food waste to the tylosin fermentation dreg for co-
digestion, the cumulative methane production increased by 14.8%–
55.5% compared to TFD alone. In the process of stable mesophilic
M. Gao et al.
anaerobic digestion, the main removal method of tylosin was the
decomposition by anaerobic microorganisms. Even if the initial con­
centration was as high as 500 mg⋅L− 1, the removal rate of tylosin could
still reach more than 68%. This study confirmed that fermentation dregs
can be co-digested with food waste in the same reactor, which has
practical significance for the anaerobic digestion of fermentation dregs.
CRediT authorship contribution statement
Ming Gao: Conceptualization, Formal analysis, Writing - original
draft. Min Yang: Methodology, Investigation, Writing - original draft.
Xinxin Ma: Visualization. Dong Xie: Investigation. Chuanfu Wu: Su­
pervision, Project administration. Qunhui Wang: Funding acquisition.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgements
This study was financially supported by the National Key R&D Pro­
gram (2018YFC1900903) and the Beijing Natural Science Foundation
Program (8192028). We would like to thank Editage (www.editage.cn)
for English language editing.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.biortech.2021.124693.
References
Cai, C., Liu, H., Wang, B., 2017. Performance of microwave treatment for disintegration
of cephalosporin mycelial dreg (CMD) and degradation of residual cephalosporin
antibiotics. J. Hazard. Mater. 331, 265–272. https://doi.org/10.1016/j.
jhazmat.2017.02.034.
Chen, W., Geng, Y., Hong, J., Jua, H.W., Xu, C., Yu, N., 2017. Life cycle assessment of
antibiotic mycelial residues management in China. Renew. Sust. Energ. Rev. 79,
830–838. https://doi.org/10.1016/j.rser.2017.05.120.
Chuenchart, W., Logan, M., Leelayouthayotin, C., Visvanathan, C., 2020. Enhancement
of food waste thermophilic anaerobic digestion through synergistic effect with
chicken manure. Biomass Bioenerg. 136, 105541. https://doi.org/10.1016/j.
biombioe.2020.105541.
Kim, J., Baek, G., Kim, J., Lee, C., 2019. Energy production from different organic wastes
by anaerobic co-digestion: Maximizing methane yield versus maximizing synergistic
effect. Renew. Energy 136, 683–690. https://doi.org/10.1016/j.
renene.2019.01.046.
Koch, K., Helmreich, B., Drewes, J.E., 2015. Co-digestion of food waste in municipal
wastewater treatment plants: effect of different mixtures on methane yield and
hydrolysis rate constant. Appl. Energy 137, 250–255. https://doi.org/10.1016/j.
apenergy.2014.10.025.
Li, C., Zhang, G., Zhang, Z., Ma, D., Wang, L., Xu, G., 2015. Hydrothermal pre-treatment
for biogas production from anaerobic digestion of antibiotic mycelial residue. Chem.
Eng. J. 279, 530–537. https://doi.org/10.1016/j.cej.2015.05.073.
Li, C., Zhang, G., Zhang, Z., Ma, D., Xu, G., 2016a. Alkaline thermal pre-treatment at mild
temperatures for biogas production from anaerobic digestion of antibiotic mycelial
residue. Bioresour. Technol. 208, 49–57. https://doi.org/10.1016/j.
biortech.2016.02.064.
Li, Y., Jin, Y., Li, J., Li, H., Yu, Z., 2016b. Effects of thermal pre-treatment on the
biomethane yield and hydrolysis rate of kitchen waste. Appl. Energy 172, 47–58.
https://doi.org/10.1016/j.apenergy.2016.03.080.
Ma, X., Yu, M., Yang, M., Gao, M., Wu, C., Wang, Q., 2019. Synergistic effect from
anaerobic co-digestion of food waste and Sophora flavescens residues at different co-
substrate ratios. Environ. Sci. Pollut. Res. 26 (36), 37114–37124. https://doi.org/
10.1007/s11356-019-06399-x.
Ma, X., Yu, M., Song, N.a., Xu, B., Gao, M., Wu, C., Wang, Q., 2020. Effect of ethanol pre-
fermentation on organic load rate and stability of semi-continuous anaerobic
View publication stats
Bioresource Technology 325 (2021) 124693
digestion of food waste. Bioresour. Technol. 299, 122587. https://doi.org/10.1016/
j.biortech.2019.122587.
Mitchell, S.M., Ullman, J.L., Teel, A.L., Watts, R.J., Frear, C., 2013. The effects of the
antibiotics ampicillin, florfenicol, sulfamethazine, and tylosin on biogas production
and their degradation efficiency during anaerobic digestion. Bioresour. Technol.
149, 244–252. https://doi.org/10.1016/j.biortech.2013.09.048.
Ni, B.-J., Zeng, S., Wei, W., Dai, X., Sun, J., 2020. Impact of roxithromycin on waste
activated sludge anaerobic digestion: Methane production, carbon transformation
and antibiotic resistance genes. Sci. Total Environ. 703, 134899. https://doi.org/
10.1016/j.scitotenv.2019.134899.
Paesen, J., Cypers, W., Busson, R., Roets, E., Hoogmartens, J., 1995. Isolation of
decomposition products of Tylosin using liquid-chromatography. J Chromatography
A 699 (1-2), 99–106. https://doi.org/10.1016/0021-9673(94)01269-K.
Rajagopal, R., Daniel, I.M., Singh, G., 2013. A critical review on inhibition of anaerobic
digestion process by excess ammonia. Bioresour. Technol. 143, 632–641. https://
doi.org/10.1016/j.biortech.2013.06.030.
Ren, Y., Yu, M., Wu, C., Wang, Q., Gao, M., Huang, Q., Liu, Y., 2018. A comprehensive
review on food waste anaerobic digestion: research updates and tendencies.
Bioresour. Technol. 247, 1069–1076. Doi: 10.1016/j.biortech.2017.09.109.
Administration, S.E.P., 2002. Water and Wastewater Monitoring and Analysis Method,
fourth ed. China Environmental Science Press, Beijing.
Stone, J.J., Clay, S.A., Zhu, Z., Wong, K.L., Porath, L.R., Spellman, G.M., 2009. Effect of
antimicrobial compounds tylosin and chlortetracycline during batch anaerobic swine
manure digestion. Water Res. 43 (18), 4740–4750. https://doi.org/10.1016/j.
watres.2009.08.005.
Wang, L.-H., Wang, Q., Cai, W., Sun, X., 2012. Influence of mixing proportion on the
solid-state anaerobic co-digestion of distiller’s grains and food waste. Biosyst. Eng.
112 (2), 130–137. https://doi.org/10.1016/j.biosystemseng.2012.03.006.
Wang, B., Li, G., Cai, C., Jiang, J., Liu, H., 2018. Assessing the safety of thermally
processed penicillin mycelial dreg following the soil application: Organic matter’s
maturation and antibiotic resistance genes. Sci. Total Environ. 636, 1463–1469.
https://doi.org/10.1016/j.scitotenv.2018.04.288.
Wu, C., Wang, Q., Yu, M., Zhang, X., Song, N., Chang, Q., Gao, M., Sonomoto, K., 2015.
Effect of ethanol pre-fermentation and inoculum-to-substrate ratio on methane yield
from food waste and distillers’ grains. Appl. Energy 155, 846–853. https://doi.org/
10.1016/j.apenergy.2015.04.081.
Wu, C., Huang, Q., Yu, M., Ren, Y., Wang, Q., Sakai, K., 2018. Effects of digestate
recirculation on a two–stage anaerobic digestion system, particularly focusing on
metabolite correlation analysis. Bioresour. Technol. 251, 40–48. https://doi.org/
10.1016/j.biortech.2017.12.020.
Yin, Y., Guo, X., Yang, C., Gao, L., Hu, Y., 2016. An efficient method for tylosin removal
from an aqueous solution by goethite modified straw mass. RSC Adv. 6 (98),
95425–95434. https://doi.org/10.1039/C6RA19172J.
Yin, F., Dong, H., Zhang, W., Zhu, Z., Shang, B., Wang, Y., 2019. Removal of combined
antibiotic (florfenicol, tylosin and tilmicosin) during anaerobic digestion and their
relative effect. Renew. Energy 139, 895–903. https://doi.org/10.1016/j.
renene.2019.03.001.
Yu, M., Gao, M., Wang, L., Ren, Y., Wu, C., Ma, H., Wang, Q., 2018a. Kinetic modelling
and synergistic impact evaluation for the anaerobic co-digestion of distillers’ grains
and food waste by ethanol pre-fermentation. Environ. Sci. Pollut. Res. 25,
30281–30291. Doi: 10. 1007/s11356-018-3027-6.
Yu, M., Wu, C., Wang, Q., Sun, X., Ren, Y., Li, Y., 2018b. Ethanol pre-fermentation of
food waste in sequencing batch methane fermentation for improved buffering
capacity and microbial community analysis. Bioresour. Technol. 248, 187–193.
https://doi.org/10.1016/j.biortech.2017.07.013.
Zhang, C., Su, H., Baeyens, J., Tan, T., 2014. Reviewing the anaerobic digestion of food
waste for biogas production. Renew. Sustain. Energy Rev. 38, 383–392. https://doi.
org/10.1016/j.rser.2014.05.038.
Zhang, G., Li, C., Ma, D., Zhang, Z., Xu, G., 2015. Anaerobic digestion of antibiotic
residue in combination with hydrothermal pre-treatment for biogas. Bioresour.
Technol. 192, 257–265. https://doi.org/10.1016/j.biortech.2015.05.014.
Zhang, B., Wang, M., Wang, B., Xin, Y., Gao, J., Liu, H., 2018. The effects of bio-available
copper on macrolide antibiotic resistance genes and mobile elements during TFDs
co-composting. Bioresour. Technol. 251, 230–237. https://doi.org/10.1016/j.
biortech.2017.12.051.
Zhang, X., Gu, J., Wang, X., Zhang, K., Yin, Y., Zhang, R., Zhang, S., 2019. Effects of
tylosin, ciprofloxacin, and sulfadimidine on mcrA gene abundance and the
methanogen community during anaerobic digestion of cattle manure. Chemosphere
221, 81–88. https://doi.org/10.1016/j.chemosphere.2018.12.043.
Zhen, G., Lu, X., Kobayashi, T., Kumar, G., Xu, K., 2016. Anaerobic co-digestion on
improving methane production from mixed microalgae (Scenedesmus sp., Chlorella
sp.) and food waste: kinetic modeling and synergistic impact evaluation. Chem. Eng.
J. 299, 332–341. https://doi.org/10.1016/j.cej.2016.04.118.
Zhong, W., Li, G., Gao, Y., Li, Z., Geng, X., Li, Y., Yang, J., Zhou, C., 2015. Enhanced
biogas production from penicillin bacterial residue by thermal-alkaline pre-
treatment. Biotechnol. Biotechnol. Equip. 29 (3), 522–529. https://doi.org/
10.1080/13102818.2015.1021277.