Journal of Cleaner Production 241 (2019) 118284

Contents lists available at ScienceDirect

Journal of Cleaner Production

journal homepage: www.elsevier.com/locate/jclepro

A new approach using an open-source low cost system for monitoring

and controlling biogas production from dairy wastewater
Ianny Andrade Cruz a, Luciano de Melo b, Ariadne Nunes Leite a,
~o Victor Melquiades Sa
Joa dia Hortense Torres c,
tiro a, Larissa Renata Santos Andrade a, Na
Rebeca Yndira Cabrera Padilla , Ram N. Bharagava , Renan Figueiredo Tavares a, c,
d e

Luiz Fernando Romanholo Ferreira a, c, *

a
Graduate Program in Process Engineering, Tiradentes University, Av. Murilo Dantas, 300, Farola^ndia, 49032-490, Aracaju, SE, Brazil
b ~o Caf
Federal Institute of Sergipe, Rod. Joa ^ncia, SE, Brazil
e Filho, 264, 49200-000, Esta
c
Institute of Technology and Research. Av. Murilo Dantas, 300, Farola^ndia, 49032-490, Aracaju, SE, Brazil
d
Federal University of Matogrosso do Sul, Av. Senador Filinto Müller, 1555, 79074-460, Campo Grande, MS, Brazil
e
Laboratory for Bioremediation and Metagenomics Research (LBMR), Department of Microbiology (DM), Babasaheb Bhimrao Ambedkar University (A
Central University), Vidya Vihar, Raebareli Road, Lucknow, 226 025, Uttar Pradesh, India

a r t i c l e i n f o a b s t r a c t

Article history: Biogas plants can effectively treat various sources of organic wastes and recover energy from biomass.
Received 26 March 2019 However, anaerobic digestion (AD) is highly sensitive to process disturbances, which can affect the biogas
Received in revised form production efficiency. Online process monitoring/controlling can be used to ensure efficient and stable
26 June 2019
biogas production, but automated instruments are still associated with high costs. Therefore, this study
Accepted 3 September 2019
developed a low-cost system, using the open-source platform Arduino and its components, to monitor/
Available online 6 September 2019
control some AD variables. To test the developed system (DS), some AD variables of the dairy wastewater
handling editor; Mingzhou Jin inoculated with sewage sludge digestion was monitored/controlled in the liquid phase (pH, temperature)
and in the gas phase (pressure, methane yield, biogas volume). The batch experiment was conducted in
Keywords: mesophilic conditions (38
C), with an inoculum/substrate ratio of 1:2 for 21 days, and it was also
Anaerobic digestion evaluated in terms of volatile solids (VS) and chemical oxygen demand (COD) removal. The DS main-
Biogas production tained the desired pH and temperature conditions and informed an average cumulative biogas and
Process monitoring/controlling methane concentration of 675.2 mL and 51.46%, respectively. The VS and COD removal rate obtained after
Low-cost system
the digestion was 45.35% and 80.1%, respectively. At the end of AD, an ecotoxicity test using Lactuca sativa
Platform arduino
seeds was performed and high digestate concentrations exhibited toxicity. Finally, the apparatus con-
Phytotoxicity
struction was feasible for the proposed work, indicating its economic potential viability.
© 2019 Elsevier Ltd. All rights reserved.

1. Introduction quality fertilizer (Deng et al., 2017; Xu et al., 2018). Furthermore, AD

Anaerobic digestion (AD) has become an attractive alternative
for energy production because of its environmental advantages
(Grando et al., 2017). During AD, various types of biomass and
organic wastes are converted into biogas (60%e70% methane, 30%e
40% carbon dioxide, and traces of others gases such as hydrogen
and ammonia), leaving a rich material that can be used as a high
* Corresponding author. Institute of Technology and Research. Av. Murilo Dantas,
^ndia, 49032-490, Aracaju, SE, Brazil.
300, Farola
E-mail addresses: romanholobio@gmail.com, luiz.fernando@souunit.com.br,
luiz_fernando@itp.org.br (L.F. Romanholo Ferreira).
process can also minimize deforestation for fire in developing
countries, take organic wastes away from landfills and replace part
of fossil fuels, hence, reducing greenhouse gases emissions and
odors (Nguyen e Khanal, 2018).
Nevertheless, AD is a complex process due to the involvement of
diverse microbial communities supporting biochemical reactions
(hydrolysis, acidogenesis, acetogenesis, methanogenesis) and it
may suffer from instability (Nguyen et al., 2015). Then, it is indis-
pensable monitoring the AD for improving its stability and effi-
ciency (Boe et al., 2010; Li et al., 2018), as well as developing cost-
effective online monitoring methods (Wu et al., 2019).
Parameters like pH, volatile fatty acids (VFA) concentrations,

https://doi.org/10.1016/j.jclepro.2019.118284
0959-6526/© 2019 Elsevier Ltd. All rights reserved.
2 I.A. Cruz et al. / Journal of Cleaner Production 241 (2019) 118284
biogas composition, biogas yield, and alkalinity are usually used as
AD indicators (Jin et al., 2017; Lützhøft et al., 2014). There are
several off-line methods for monitoring AD both liquid and gas
phases such as gas chromatography (GC) (Ward et al., 2011), pH
titration (Lützhøft et al., 2014), high performance liquid chroma-
tography (HPLC) and mid-infrared spectroscopy (Falk et al., 2015).
However, these methods are expensive, time consuming and
commonly tested manually. There are also some studies of online
monitoring systems based on the above-named methods (Boe e
Angelidaki, 2012), but they usually require complex equipment
and meticulous maintenance, or even a challenging pretreatment.
Biolectrochemical systems have become an alternative for the on-
line monitoring of these processes (Jin et al., 2017), but the devel-
opment of efficient and cost-effective biosensors still needs
improvements.
Yu et al. (2016) state that the gas-liquid phase monitoring makes
possible indicating the digester state by simple parameters such as
the liquid-phase pH and biogas flow rate of the gas-phase, so they
developed a system which combined biogas-pH monitoring and
controlling thresholds. While Bernardi et al. (2017) developed a
medium scale plant which monitor in real time the system tem-
perature, pH, biogas and subsequently the methane through gas
chromatograph. The above studies used programmable logic con-
trollers (PLC), which is more expensive and usually used in in-
dustries, but it is possible to efficiently monitor the AD process
using low-cost and accessible electronics and the platform Arduino.
The Arduino is an open-source electronic platform which allows
the building of scientific instruments, using the open-source
hardware and software, and reducing the cost of researches
(Maraba e Bulur, 2017). It has been efficiently used to monitor and
control experimental equipment in automation processes by the
scientific community. Among the various studies that have used it,
Mesas-Carrascosa et al. (2015) performed the temperature and
relative humidity of the air monitoring in agriculture, Resende et al.
(2017) constructed a low-cost plant for biodiesel production, Rosa
et al. (2017) developed an apparatus that controls an electro-
chemical pretreatment, and Ahmed et al. (2017) developed a sys-
tem to measure only the methane production in the AD process,
which is different of the present study that monitors more vari-
ables, such as pressure, biogas volume, pH and temperature.
Therefore, one of the shortcomings of AD technologies is the
high capital cost associated with reactors which are usually made of
expensive materials (Budzianowski, 2016). Besides studies state
that process automation and control should be integrated as part of
the bioreactor design (Khanal et al., 2017), instrumentation (in both
hardware and software aspects) is still associated with high costs
(Maraba e Bulur, 2017). Thus, this study used the platform Arduino
and some low-cost electronic components, to construct an auto-
mated system capable of online monitoring/controlling some in-
dicators of the AD process in the liquid phase (pH, temperature)
and in the gas phase (pressure, methane yield, biogas volume).
Furthermore, the AD of dairy wastewater inoculated with sewage
sludge was performed, and at the end of the digestion period, a
toxicity assessment of the digestate was realized using the bio-
indicator Lactuca sativa.
2. Materials and methods
2.1. Apparatus construction
This plant is composed of a Mariotte vessel having a capacity of
about 2 L (working volume), closed with rubber stoppers in which
five holes were open, in order to attach the pH and temperature
sensors, to allow biogas output and to add an acid or basic solution
(pH control). Heating is ensured by a thermostatically regulated
heater, that goes on/off whenever temperature exceeds the set
thresholds, holding the vessel temperature in the desired range.
And the reactor mixing is accomplished by a system of magnetic
stirring which offers a mixing velocity of 100e1500 rpm and is
manually adjustable.
The digester is also provided by a temperature sensor (DS18B20)
that control the heater functioning and a suitable pH probe (Arduo
eletro) with a transducer for Arduino that control the peristaltic
pumps to add an acid or basic solution.
Pressure is checkable due to a differential pressure sensor
(MPX5050) at the biogas output line. Moreover, the software is
provided by a series of biogas releases and delays in order to pre-
vent critical overpressure or under pressure. Then, the produced
biogas passes by a non-dispersive infrared sensor (CH4/GP) that
enable the methane concentration measurement. Finally, the
biogas goes to a receptor vessel which is above a load sensor (AF)
sensible for strain gauge that informs the produced biogas volume.
Despite the temperature sensor has been already calibrated in
its fabrication, the other sensors (pH, pressure, load and methane)
were calibrated in this research. Some standard solutions with
known pH were used to calibrate the pH sensor and obtain its
response values. To obtain the response values of the pressure and
load sensor, different pressures were applied and monitored by a
manometer, and different standard weights were used, respec-
tively. Finally, to calibrate the methane sensor, a sample of CH4
standard gas (99,98%) was injected into the sensor, and its response
values obtained. The obtained response values for each sensor were
used to construct scatterplots, and then, obtain equations of the
type y ¼ ax þ b (linear regression). Statistically, for each curve, a
discrepancy of 5% represented by the variation coefficient was
accepted, above that the values were discarded, and the procedure
redone. Thus, all equations were added in the software and
uploaded in the Arduino, so all the values obtained by the sensors
are presented in their respective units.
Some Arduino-compatible modular electronics known as
Nanoshields (Circuitar) are also used, because they make the sys-
tem more robust. The ADC 4e20 Nanoshield, for example, allows to
measure the methane sensor with 4 mAe20 mA output with high
precision and resolution. The Load Cell Nanoshield is connected to
the load sensor and permits high-precision and high-resolution
load cell measurement. With the MicroSD Nanoshield is possible
to record all the data obtained in a microSD card, and with the
RTCMem Nanoshield a real time clock is added to the system.
The system software is written in the Arduino Integrated
Development Environment (IDE) and is responsible for incorpo-
rating multifunction of equipment automation control, data
acquisition, recording and display. In addition, a liquid crystal
display (LCD), some light-emitting diodes (LEDs) and a sound sys-
tem are also implemented to improve the user experience. The
system is also equipped with a USB communication interface that
provides the data export process to a Personal Computer (PC). The
schematic diagram of the reactor used in this study is shown in
Fig. 1.
During the process, effluents samples can be collected, as well as
at the end of the process, the vessel drain can be effectuated by
means of a manual valve present in the bottom.
2.2. Functioning
As previously mentioned, temperature and pH are the main
parameters that control the prototype automatism. So, feeding runs
if the pH value goes beyond the set thresholds, loading so, alkaline
feed or acid one according to the registered pH increase or decrease.
Analogously, substrate heating is controlled by a temperature
probe, then, if substrate temperature goes bellow the set value the
 I.A. Cruz et al. / Journal of Cleaner Production 241 (2019) 118284
Fig. 1. Flow and logic diagram of the experimental setup.
thermostats run turning the heating on, otherwise, they go off until
the vessel cooling.
The produced biogas is determined by means of volume
displacement (Moukazis et al., 2017), done automatically using a
bottle containing a liquid above the load sensor. So, as far as the
biogas is produced, it goes to a receptor vessel that is full of a liquid
and expel it; the load sensor measures the expelled liquid mass and
thanks to the density equation saved in the firmware, the system
presents the expelled liquid volume which is the same of the biogas
produced. Methane quantity included in the biogas is measured,
through a quantitative analysis, due to the CH4 sensor that informs
its concentration in percentage.
All the monitoring data is collected by the Arduino, then
recorded in a microSD card and showed in a liquid crystal display
(LCD) in real-time. The process record can be extracted to the PC at
any time.
2.3. Raw material selection and inoculation
In this study it was used a dairy wastewater (DW) from a cheese
factory located in Campo do Brito/SE (10º440 38.000 S 37º290 42.300 W)
which uses traditional technologies for cheese manufacture. The
DW characteristics are shown in Table 1.
For the anaerobic digestion process startup, anaerobic sludge
(AS) from a municipal wastewater treatment plant located in Barra
dos Coqueiros/SE (10º530 58.800 S 37º01057.500 W) has been used. This
inoculum was collected, sieved (<5 mm) to remove larger particles
and stored at 4
C in a cold chamber until its use. The properties of
the inoculum used for this study are presented in Table 2.
Table 1
Main characteristics of dairy wastewater.
Analysis Units
pH e 4.48
Total solids (TS) (mg/L)
Volatile solids (VS) (mg/L)
Chemical Oxygen Demand (COD) (mgO2/L)
Biochemical Oxygen Demand (BOD) (mgO2/L)
Oil and grease (mg/L)
Total nitrogen (mg/L)
Total phosphorus (mg/L)
Sulfates (mg/L)
Potassium (mg/L)
3
Table 2
Characterization of the inoculum used in the experiments.
Analysis Inoculum
Total solids (mg/L) 915
Volatile solids (mg/L) 270
Chemical Oxygen Demand (mg/L) 1985.5
pH value 6.92
Temperature (oC) 30
2.4. Characterization methods
The characterization of the feed, the reactive mixture and the
biogas were performed according to the following methods: total
solids (TS), volatile solids (VS), total chemical oxygen demand
(COD), total Kjeldahl nitrogen (TKN) and fat content were deter-
mined in accordance with the Standard Methods for the Exami-
nation of Water and Wastewater (Apha, 2012).
Temperature and pH were measured offline using a pH meter
Spenccer and online using a sensor (DS18B20) and an electrode
(Arduo Eletro), respectively.
Ecotoxicological bioassays were carried out according to meth-
odology proposed by Vilar et al. (2018).
2.4.1. Ecotoxicological assays using Lactuca sativa
In order to assess the toxicological response, lettuce seeds (Lac-
tuca sativa) were used as toxicity level bioindicators. The biotests
applied in this paper were developed according to the methodology
proposed by Vilar et al. (2018). Three replicates were tested on a set of
dilutions (12.5%, 25%, 50%, 75%, 100%, containing 20 lettuce seeds
each) using the raw dairy wastewater and the digestate (after the
anaerobic digestion) samples. Negative control with 0.2 M NaCl so-
lution and a positive control with distilled water were used. Using
Petri plates, the lettuce seeds were put on a layer of filtration-type
paper containing solutions (samples or distilled water) and prop-
erly covered by aluminum foil to maintain it in dark conditions. Then,
they were germinated inside an incubator at constant temperature of
22 ± 2
C. The germination was registered after 120 h of exposure and
germination rate (GR) calculated. The GR% is determined using the
Equation (1), where SGA is the number of seeds germinated in the
sample and SGC is the number of seeds germinated in the control.
N
SGA
%GR ¼ *100 (1)
N
SGC
2.5. Experimental procedure
Prior to incubation, the reaction vase was flushed with nitrogen
to ensure an anaerobic ambient. Then, the inoculum and substrate
were mixed to the required ratio of 1:2 (Koch et al., 2017) and
inoculated in the reactor manually. It operated at mesophilic tem-
perature conditions (approximately 38
C) with a hydraulic reten-
tion time of about 20 days (Kumari et al., 2018), with agitation set at
300 rpm. The pH was automatically adjusted to 6.5e7.8 by adding
Values solutions of 1M NaOH and 4M HCl through peristaltic pumps, as
ideal condition for methanogenic bacteria (Choong et al., 2017).
52 640 Besides that, effluent samples were collected twice a week
46 465 during the experiments for analyzing pH, COD, TS and VS.
78 057.3
42 650
820 3. Results and discussion
1200
369.9 3.1. Equipment monitoring
276
4.56
To evaluate the DS economic viability, it was made a quotation
4 I.A. Cruz et al. / Journal of Cleaner Production 241 (2019) 118284

request of a commercial bioreactor (CB) with same functionalities,
such as a set of peristaltic pumps, a measurement and actuation
device, a reaction vessel and its support, temperature controlling,
an agitation mechanism, connectors and a control software. The
final value of it was approximately 25 000.00 US dollars, but it is
necessary to point out some additional characteristics of the CB as
its vessel capacity of 7 L, an aeration module, and that is made of
glass and stainless steel. Nevertheless, to add a biogas (CH4, CO2, O2
and H2S) analyzer module in the CB, its price would increase to
approximately 34 000 US dollars.
Nonetheless, the purchase of all components cited in Fig. 1, and
the DS total construction had an average cost of 1560.00 US dollars.
It is worth to note that the CB may have a higher efficiency, but for
academic purposes the DS shows a better economic viability.
3.1.1. pH
pH is a very important parameter in the anaerobic digestion
because it shows the process stability and performance (Kumari
et al., 2018). The anaerobic digestion process includes four steps
namely hydrolysis, acidogenesis, acetogenesis and methanogenesis
and the bacteria present in these stages largely depend on system
pH where anaerobic digestion occurs (Mao et al., 2015).
The ideal pH range in the AD process is between 6.5 and 7.8
which is sufficient for the survival of methanogenic bacteria and

(a)

8.00

7.00

6.00

5.00
pH

4.00

3.00
Online method (DS)
2.00
Offline method
1.00

0.00
0 5 10 15 20
Time, days

(b)

40.0

39.0
Temperature, oC

38.0

37.0
Online method (DS)
Offline method
36.0
0 5 10 15 20
Time, days
Fig. 2. pH (a) and temperature (b) variation of the effluent during the digestion period (Mean values of three independent replications; vertical bars represent standard errors).
 I.A. Cruz et al. / Journal of Cleaner Production 241 (2019) 118284
organic matter degradation (Choong et al., 2017). However, due to
the dairy wastewater acidic and oily nature which resulted in more
acid formation, the pH was initially in the range of 5e6, how Fig. 2
(a) shows. Aiming to avoid a pH larger decrease, 1M NaOH has been
used as the alkali solution utilized for stabilizing the pH value just
as carried out by Streitwieser (2017). In their study, Kumari et al.
(2018) evaluated the co-digestion of dairy wastewater with cow
manure, which had an initial pH of 6.86, indicating that its pH was
sufficient for bacteria survival, and inhibition by acidification was
not a problem. Sivakumar et al. (2012), had to adjust the influent
(spoiled milk) pH by adding aqueous solutions of sodium bicar-
bonate, and notticed that lower pH resulted in lower volume biogas
production, which indicates the need of the pH controlling.
Observing Fig. 2 (a), it is also possible to see that up to day 11 the
pH was between 5 and 6, which can be interpreted as the AD initial
stages (hydrolysis, acidogenesis, acetogenesis). However, from day
12 the pH started to increase, reaching the ideal range for meth-
anogenic bacteria and, as can be seen in Fig. 4, it is during this
period that methane production actually occurred, thus indicating
the process methanogenic phase.
Every time samples were collected from the reactor, pH of them
was measured in a common pH meter (Spenccer) with intention of
comparing these values with those obtained by the DS, in order to
demonstrate the equipment efficiency. In Fig. 2 (a) it is also possible
to see the comparison between these values which were noticeably
close.
3.1.2. Temperature
AD processes can be accomplished over three temperature
ranges: psychrophilic (10e30
C), mesophilic (20e50
C) and
thermophilic (35e75
C) (Leite et al., 2017). The treatment becomes
more difficult as temperature drops below 20
C (Bowen et al.,
2014), so mesophilic and thermophilic are preferred in most
anaerobic experiments, once higher temperature may lead to a
higher biogas production (Leung e Wang, 2016). However, ther-
mophilic AD has some disadvantages such as decreased stability,
larger investments and higher net energy inputs, what makes the
mesophilic digestors more common, mainly in tropical regions
(Mao et al., 2015).
The optimum temperature of digestion can vary depending on
the effluent composition, but it is important to maintain it at an
approximately constant level for achieving a maximum gas pro-
duction rate (Kothari et al., 2014). Therefore, it is necessary to
control the temperature during the AD process, and the built
800
Cumula ve
700
Cumula ve biogas yield, mL
Daily
600
500
400
300
200
100
0 0
0 5
Time, days
Fig. 3. Cumulative and daily yields during anaerobic digestion of dairy wastewater.
5
system was able to do it as Fig. 2 (b) shows.
The temperature of aliquots collected in the reactor was also
measured outside the system and the results are shown in Fig. 2 (b)
just like happened for pH, the values compared were noticeably
close for temperature. However, it is important to note that the
environment can influence in the measurement results, making the
system results more reliable.
3.1.3. Biogas and methane yield
The cumulative and daily biogas yield are shown in Fig. 3. On the
first day, one peak of daily biogas yield (1.378 mL/day) was
observed, due to the dairy wastewater higher COD content that
could be quickly biodegraded in the first day. However, biogas
production rate dropped after this peak, and no biogas was pro-
duced from day 3e12, indicating an apparent severe inhibition. This
biogas production inhibition was likely caused by the substrate
high digestibility that led to volatile fatty acids (VFA) over-
production and inhibited the methanogenesis process (Abudi et al.,
2016). After 9 days of methanogenic bacteria self-recovery, the
biogas production started again. From Fig. 3, it also can be seen that
at the end of the digestion process, the total cumulative biogas yield
was 675.2 mL.
In Fig. 4 can be noticed that the methane production started
after the 14th day, with the highest peak of methane content
(51.46%) at the 21st day of digestion. In their study, Lovato et al.
(2016) got 79.1% of methane content in the co-digestion of syn-
thetic cheese whey and glycerin, while the present work just used
the real dairy wastewater without glycerin addition, which could
lead to a higher biogas production.
3.2. COD removal during AD process
COD indicates the measure of organic pollutants present in the
sample (Kumari et al., 2018). COD concentration with respect to
time is shown in Fig. 5. The decrease in the COD concentration
during the anaerobic digestion was observed, and it represents a
decrease in the dairy wastewater organic pollutant. At the end of 21
days digestion period, the process obtained a COD removal effi-
ciency of 80.1%. In their studies, Kumari et al. (2018) obtained a COD
removal of 86.56% after the co-digestion of dairy wastewater and
cow manure at mesophilic temperature, and Lovato et al. (2016)
achieved 89% of COD removal during the co-digestion of syn-
thetic cheese whey and glycerin. It is worth noting that in study
carried out by Kumari et al. (2018) an inoculum composed of bovine
2.5
Daily biogas yield, mL/day
2
1.5
1
0.5
10 15 20
6 I.A. Cruz et al. / Journal of Cleaner Production 241 (2019) 118284

80.00
% CH4
70.00

Methane concentra on, %

60.00
50.00
40.00
30.00
20.00
10.00
0.00
0 5 10 15 20
Time, days

Fig. 4. Biogas methane concentration obtained by the built system using the NDIR sensor (Mean values of three independent replications; vertical bars represent standard errors).

50,000
45,000
COD concentra on, mg/L

40,000 COD
35,000
30,000
25,000
20,000
15,000
10,000
5,000
0
0 5 10 15 20 25
Time, days

Fig. 5. COD concentration with time for the effluent (dairy wastewater þ anaerobic sludge) coming out of the reactor (Mean values of three independent replications; vertical bars
represent standard errors).

manure was used, which contains a different microbial diversity
than the used in the present work, which modifies the process
efficiency of COD removal. While Lovato et al. (2016) worked with a
synthetic cheese whey plus glycerin, characterized by lower COD
(5000 mgCOD/L) when compared with the real effluent, which could
explain the greater COD reduction.
they obtained a VS removal of 17.4% and 16%, respectively, which
indicates an increase in efficiency when the co-digestion is per-
formed, what did not happen in the present work which used just
one substrate (mono-digestion).
3.4. Toxicity

3.3. Volatile solids removal
The reduction in volatile solids (VS) is one of the most valuable
parameters for evaluating the AD efficiency and it indicates the
substrate biodegradability (Abudi et al., 2016; Leung e Wang, 2016).
As shown in Fig. 6 the VS concentration had a decrease and at the
end of the process 45.35% VS removal has been reached through the
mono-digestion of dairy wastewater. Pavi et al. (2017) reported
54.6% as the highest rate of VS removal in the co-digestion of
organic fraction of municipal solid waste (OFMSW) and fruit and
vegetable waste (FVM), but in mono-digestion of OFMSW and FVW,
Besides biogas, the AD also has as product a residue called
digestate which is a mixture of partially degraded organic matter,
inorganic compounds and microbial biomass (Alburquerque et al.,
2012). This digestate can be used as biofertilizer in agriculture,
but its direct application may be limited because it may contain
heavy metals, infrequent parasites and seeds or weeds (Stefaniuk
et al., 2016). So, ecotoxicity analyses of digestate before its use in
agriculture can predict its environmental impact or a necessity for
additional treatments (Tigini et al., 2016). Fig. 7 shows the results
for the ecotoxicological test made using Lactuca sativa seeds.
According to a study from Cesaro et al. (2015), analyzing organic
 I.A. Cruz et al. / Journal of Cleaner Production 241 (2019) 118284 7

35,000

30,000

Vola e Solids, mg/L

25,000

20,000

15,000

10,000
VS
5,000

0
0 5 10 15 20 25
Time, days

Fig. 6. Volatile solids (VS) concentration with time for the effluent (dairy wastewater þ anaerobic sludge) coming out of the reactor (Mean values of three independent replications;
vertical bars represent standard errors).

(a) (b)

(c) (d)

Fig. 7. Dairy wastewater germination index (a) and relative root growth (b). Initial mixture (dairy wastewater þ sewage sludge) and digestate germination index (c) and relative
root growth (d).
8 I.A. Cruz et al. / Journal of Cleaner Production 241 (2019) 118284
residues phytotoxicity using L. sativa seeds, all results for a germi-
nation index (GI) below 40% indicate seed inhibition sensibility, the
values between 80 and 120% are not considered significant, and the
values above 120% are considered as growing stimulus. Therefore,
the results from this study indicate no toxicity for the dairy
wastewater in natura, as can be seen in Fig. 7 (a), while Fig. 7 (b)
shows a slightly growth (10% higher) when compared to the posi-
tive control, which evidences the wastewater performance as
inducer in the root growth.
In Fig. 7 (c), no germination was observed for the initial mixture
(dairy wastewater þ sewage sludge) concentrations equal or
greater than 75%, nor for digestate concentrations equal or greater
than 50%. The seed germination index around 70% indicates a slight
inhibition in the seed germination for initial mixture concentra-
tions of 12.5%, 25% and 50%. However, the initial mixture concen-
trations of 12.5% and 25% served as inducer to the root growth, as
can be seen in Fig. 7 (d), whereas the 50% concentration presented a
growth lower than the one presented by the positive control. This
slight inhibition in the seed germination and the reduced growth,
for the 50% mixture concentration, may be explained by the pres-
ence of toxic substances in the sewage sludge that was used as
inoculum.
For the digestate concentrations of 12.5% and 25%, the germi-
nation index was around 80% which would be characterized as a
non-significant value for germination. However, when looking at
Fig. 7 (d) it is noted that root growth decreased gradually until the
absence of growth in the 50% concentration of digestate. This
episode may have occurred due to the presence of some AD
products, Khanal et al. (2017), for example, state that in the AD of
some industrial effluents, such as dairy, there is ammonia genera-
tion in high concentration, which could have affected the root
growth.
Toxicity studies for dairy wastewater are scarce in literature, so
this is an important report for this effluent to study the phytotox-
icity effects using the L. sativa seeds. But Tigini et al. (2016) evalu-
ated pig slurry digestate through some ecotoxicity tests and found
out that it was very toxic and needed a pretreatment before its use
as fertilizer. This result was similar to the one obtained in the
present work, as can be seen in Fig. 7 (c, d), where higher per-
centages of the digestate resulted in no germination, indicating the
need of some pretreatment before the use of high digestate con-
centrations as biofertilizer.
4. Conclusions
A system to monitor and control some variables of AD process
was developed using some low acquisition cost sensors, actuators
and the platform Arduino. This proposed system was designed as
an alternative to higher cost equipment, improving the biogas
production quality and decreasing management costs. The moni-
toring and control processes kept the desired pH and temperature
conditions, demonstrating satisfactory results, and the total cu-
mulative biogas yield achieved was 675.2 mL.
It is critical to perform more tests for comparing the developed
system efficiency with commercial reactors ones, as well as making
the system scale up. Furthermore, it is interesting to monitor more
variables, such as volatile fatty acids and gases like CO2 and H2S, as
way of monitoring more closely the anaerobic digestion process
and enhance the biogas production. Besides that, a deeper eco-
nomic feasibility analysis is important to motivate future imple-
mentation of the developed system.
Acknowledgments
The authors would like to thank the Institute of Technology and
Research at Tiradentes University (ITP-UNIT), the Foundation of
Support to Research and Technological Innovation of the State of
Sergipe [FAPITEC/SE], the Coordination for the Improvement of
Higher Education Personnel (CAPES) under the Finance Code 001
and the National Council for Scientific and Technological Devel-
opment (CNPq) for financial support (process no 315405/2018e0;
421147/2016e4; 445203/2014-5).
References
Abudi, Z.N., Hu, Z., Sun, N., Xiao, B., Rajaa, N., Liu, C., Guo, D., 2016. Batch anaerobic
co-digestion of OFMSW (organic fraction of municipal solid waste), TWAS
(thickened waste activated sludge) and RS (rice straw): influence of TWAS and
RS pretreatment and mixing ratio. Energy 107, 131e140.
Ahmed, W.A., Aggour, M., Naciri, M., 2017. Biogas Control : methane production
monitoring using arduino. Int. J. Biotechnol. Bioeng. 11 (2), 130e133.
Alburquerque, J.A., Fuente, L.A., De, C., Ferrer-Costa, A., Carrasco, L., Cegarra, J.,
Abad, M., Bernal, M.P., 2012. Assessment of the fertiliser potential of digestates
from farm and agroindustrial residues. Biomass Bioenergy 40, 181e189.
Apha. Standard Methods for the Examination of Water and Wastewater. ([s.l: s.n.]).
Bernardi, B., Benalia, S., Zema, A.D., Tamburino, V., Zimbalatti, G., 2017. An auto-
mated medium scale prototype for anaerobic co-digestion of olive mill waste-
water. Info. Process. Agri. 4 (4), 316e320.
Boe, K., Angelidaki, I., 2012. Pilot-scale application of an online VFA sensor for
monitoring and control of a manure digester. Water Sci. Technol. 66 (11),
2496e2503.
Boe, K., John, D., Steyer, J., Angelidaki, I., 2010. State indicators for monitoring the
anaerobic digestion process. Water Res. 44 (20), 5973e5980.
Bowen, E.J., Dol, J., Davenport, R.J., Read, F.L., Curtis, T.P., Curtis, T.P., 2014. Low-
temperature Limitation of Bioreactor Sludge in Anaerobic Treatment of Do-
mestic Wastewater. Water Science & Technology, pp. 1004e1013.
Budzianowski, W.M., 2016. A review of potential innovations for production, con-
ditioning and utilization of biogas with multiple-criteria assessment. Renew.
Sustain. Energy Rev. 54, 1148e1171.
Cesaro, A., Belgiorno, V., Guida, M., 2015. Compost from organic solid waste: quality
assessment and European regulations for its sustainable use. Resour. Conserv.
Recycl. 94, 72e79.
Choong, Y.Y., Chou, K.W., Norli, I., 2017. Strategies for improving biogas production
of palm oil mill effluent (POME) anaerobic digestion: a critical review. Renew.
Sustain. Energy Rev. 82, 2993e3006. January.
Deng, L., Liu, Y., Zheng, D., Wang, L., Pu, X., Song, L., Wang, Z., Lei, Y., Chen, Z.,
Long, Y., 2017. Application and development of biogas technology for the
treatment of waste in China. Renew. Sustain. Energy Rev. 70, 845e851.
February.
Falk, H.M., Reichling, P., Andersen, C., Benz, R., 2015. Online monitoring of con-
centration and dynamics of volatile fatty acids in anaerobic digestion processes
with mid-infrared spectroscopy. Bioproc. Biosyst. Eng. 38 (2), 237e249.
Grando, R.L., Souza Antune, A.M.D.E., Fonseca, F.V.D.A., S anchez, A., Barrena, R.,
Font, X., 2017. Technology overview of biogas production in anaerobic digestion
plants: a European evaluation of research and development. Renew. Sustain.
Energy Rev. 80, 44e53. February.
Jin, X., Li, X., Zhao, N., Angelidaki, I., Zhang, Y., 2017. Bio-electrolytic sensor for rapid
monitoring of volatile fatty acids in anaerobic digestion process. Water Res. 111,
74e80.
Khanal, S.K., Giri, B., Nitayavardhana, S., Gadhamshetty, V., 2017. Anaerobic Bio-
reactors/Digesters: Design and Development. Elsevier B.V. [s.l.].
Koch, K., Lippert, T., Drewes, J.E., 2017. The role of inoculum's origin on the methane
yield of different substrates in biochemical methane potential (BMP) tests.
Bioresour. Technol. 243, 457e463.
Kothari, R., Pandey, A.K., Kumar, S., Tyagi, V.V., Tyagi, S.K., 2014. Different aspects of
dry anaerobic digestion for bio-energy: an overview. Renew. Sustain. Energy
Rev. 39, 174e195.
Kumari, K., Suresh, S., Arisutha, S., Sudhakar, K., 2018. Anaerobic co-digestion of
different wastes in a UASB reactor. Waste Manag. 77, 545e554.
Leite, W., Magnus, B.S., Guimar~ aes, L.B., Gottardo, M., Belli Filho, P., 2017. Feasibility
of thermophilic anaerobic processes for treating waste activated sludge under
low HRT and intermittent mixing. J. Environ. Manag. 201, 335e344. November.
Leung, D.Y.C., Wang, J., 2016. An overview on biogas generation from anaerobic
digestion of food waste. Int. J. Green Energy 13 (2), 119e131.
Li, L., Peng, X., Wang, X., Wu, D., 2018. Anaerobic digestion of food waste: a review
focusing on process stability. Bioresour. Technol. 248, 20e28, 174.
LOVATO, G., RATUSZNEI, S.M., RODRIGUES, J.A.D., 2016. Co-digestion of whey with
glycerin in an AnSBBR for biomethane production. Appl. Biochem. Biotechnol.
178, 126e143.
Lützhøft, H.C.H., Boe, K., Fang, C., Angelidaki, I., 2014. Comparison of VFA titration
procedures used for monitoring the biogas process. Water Res. 54, 262e272.
Mao, C., Feng, Y., Wang, X., Ren, G., 2015. Review on research achievements of
biogas from anaerobic digestion. Renew. Sustain. Energy Rev. 45, 540e555.
Maraba, D., Bulur, E., 2017. Design and construction of an automated OSL reader
with open source software and hardware. Radiat. Meas. 106, 632e637.
Mesas-Carrascosa, F.J., Verdú Santano, D., Meron ~ o, J.E., Sa
nchez De La Orden, M.,
García-Ferrer, A., 2015. Open source hardware to monitor environmental
 I.A. Cruz et al. / Journal of Cleaner Production 241 (2019) 118284
parameters in precision agriculture. Biosyst. Eng. 137, 73e83.
Moukazis, I., Pellera, F.-M., Gidarakos, E., 2017. Slaughterhouse By-Products Treat-
ment Using Anaerobic Digestion. Waste Management.
Nguyen, D., Gadhamshetty, V., Nitayavardhana, S., Kumar, S., 2015. Automatic pro-
cess control in anaerobic digestion technology : a critical review. Bioresour.
Technol. 193, 513e522.
Nguyen, D., Khanal, S.K., 2018. A little breath of fresh air into an anaerobic system:
how microaeration facilitates anaerobic digestion process. Biotechnol. Adv. 36,
1971e1983.
Pavi, S., Kramer, L.E., Gomes, L.P., Miranda, L.A.S., 2017. Biogas production from co-
digestion of organic fraction of municipal solid waste and fruit and vegetable
waste. Bioresour. Technol. 228, 362e367.
Resende, I.T.F., Coriolano, D.L., Andrade, V.C.V., Gonçalves, D.P., Oliveira, A.C.M.,
Santana, H.N., Santos, A.J., Figueiredo, R.T., 2017. Biodiesel production by ther-
mal solar heating in a low cost plant using waste cooking oil. Int. J. Machanica.
Eng. Robotics. Res. 6 (2), 160e163.
Rosa, T.R., Betim, F.S., Ferreira, R.D.E.Q., 2017. Development and application of a
labmade apparatus using open-source “arduino” hardware for the electro-
chemical pretreatment of boron-doped diamond electrodes. Electrochim. Acta
231, 185e189.
Sivakumar, P., Bhagiyalakshmi, M., Anbarasu, K., 2012. Anaerobic treatment of
spoiled milk from milk processing industry for energy recovery e A laboratory
to pilot scale study. Fuel 96, 482e486.
Stefaniuk, M., Oleszczuk, P., Bartmin  ski, P., 2016. Chemical and ecotoxicological
9
evaluation of biochar produced from residues of biogas production. J. Hazard
Mater. 318, 417e424.
Streitwieser, D.A., 2017. Comparison of the anaerobic digestion at the mesophilic
and thermophilic temperature regime of organic wastes from the agribusiness.
Bioresour. Technol. 241, 985e992.
Tigini, V., Franchino, M., Bona, F., Varese, G.C., 2016. Is digestate safe? A study on its
ecotoxicity and environmental risk on a pig manure. Sci. Total Environ.
551e552, 127e132.
Vilar, D.S., Carvalho, G.O., Pupo, M.M.S., Aguiar, M.M., Torres, N.H., Ame rico, J.H.P.,
Cavalcanti, E.B., Eguiluz, K.I.B., Salazar-Banda, G.R., Leite, M.S., Ferreira, L.F.R.,
2018. Vinasse degradation using Pleurotus sajor-caju in a combined biological e
electrochemical oxidation treatment. Separ. Purif. Technol. 192, 287e296.
Ward, A.J., Bruni, E., Lykkegaard, M.K., Feilberg, A., Adamsen, A.P.S., Jensen, A.P.,
Poulsen, A.K., 2011. Real time monitoring of a biogas digester with gas chro-
matography , near-infrared spectroscopy , and membrane-inlet mass spec-
trometry. Bioresour. Technol. 102 (5), 4098e4103.
Wu, D., Li, L., Zhao, X., Peng, Y., Yang, P., Peng, X., 2019. Anaerobic digestion : a
review on process monitoring. Renew. Sustain. Energy Rev. 103, 1e12.
Xu, F., Li, Y., Ge, X., Yang, L., Li, Y., 2018. Anaerobic digestion of food waste e chal-
lenges and opportunities. Bioresour. Technol. 247, 1047e1058. September.
Yu, D., Liu, J., Sui, Q., Wei, Y., 2016. Biogas-pH automation control strategy for
optimizing organic loading rate of anaerobic membrane bioreactor treating
high COD wastewater. Bioresour. Technol. 203, 62e70.