Journal of Environmental Management 241 (2019) 587–602

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Journal of Environmental Management

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

Research article

ADM1-based mechanistic model for the role of trace elements in anaerobic T

digestion processes
L. Frunzoa,∗, F.G. Fermosob, V. Luongoa, M.R. Matteia, G. Espositoc
a
Department of Mathematics and Applications “Renato Caccioppoli”, University of Naples Federico II, via Cintia I, Monte S. Angelo, 80126, Naples, Italy
b
Instituto de la Grasa (C.S.I.C.), Campus Universidad Pablo de Olavide, Edificio 46, Ctra. de Utrera km. 1, 41013, Sevilla, Spain
c
Department of Civil, Architectural and Environmental Engineering, University of Naples Federico II, via Claudio 21, 80125, Naples. Italy

ARTICLE INFO ABSTRACT

Keywords: An original mechanistic model able to describe the fate of trace elements (TE) in anaerobic digestion systems has
Trace elements been synthetized from mass balance equations. The model takes into account the main biochemical and physico-
Anaerobic digestion chemical processes affecting TE bioavailability and it is aimed at evaluating the effect that the combination of
Mathematical modelling such processes exerts on the system performance. Five main modules have been introduced: biochemistry,
Ordinary differential equations
physico-chemistry, sorption, complexation and precipitation. The model is based on mass conservation princi-
ples and is formulated as a set of ordinary differential equations for the soluble and particulate components
constituting the system. Model applications of two illustrative cases are provided. The first case is based on
experimental results and examines the effect of TE depletion in an AD process of food waste (FW). The second
case shows the effects of different metal supplements on methane production and biogas composition. The
simulation results confirm that the model can fairly be used to predict the effect of TE dynamics and bioa-
vailability, by considering biological, chemical and physicochemical processes in AD environments.

1. Introduction Fermoso et al., 2015; Ortner et al., 2014).

Anaerobic Digestion (AD) has been widely used to treat high volatile
solids (VS) content substrates and results in substrate stabilization and
high net energy recovery (Esposito et al., 2012). Optimal supply of
Trace Elements (TE) is a prerequisite for microbial growth and AD
microbial metabolism. Consequently, the deficiency of TE leads to
significant limitation of microbial activity that can result in the com-
plete failure of AD process (Van Hullebush et al., 2016). TE such as iron
(Fe), cobalt (Co), nickel (Ni), selenium (Se), molybdenum (Mo) and/or
tungsten (W), play a key role in the growth of anaerobic microorgan-
isms, being part of essential enzymatic co-factors such as methyl-
coenzyme M, carbon monoxide dehydrogenase, or coenzyme M methyl-
transferase, involved in the AD processes (Zhang et al., 2015a).
TE dynamics and bioavailability in AD systems are affected by
biological, chemical and physical processes. TE are subjected to com-
plex interactions, with the biomass and the liquid phase (e.g., pre-
cipitation, organic complexation, bio-sorption, adsorption or inorganic
complexation), which affect the TE bio-uptake and the process stability.
Predicting the fate of TE in anaerobic processes has received increased
attention across the research community (Van Hullebush et al., 2016;
The definition of a mathematical model able to consider the com-
plex dynamics and bioavailability of TE in anaerobic environment is
highly relevant for both academia and industries. It can allow the un-
derstanding of the main processes affecting TE in AD systems while
providing an user-friendly tool for TE requirements in full-scale appli-
cations.
In 2002 the International Water Association (IWA) Task Group for
Mathematical Modelling of Anaerobic Digestion Processes developed a
comprehensive mathematical model known as Anaerobic Digestion
Model 1 (ADM1) (Batstone et al., 2002). The ADM1 is a well-known and
widely studied mathematical model, which can describe the conversion
of complex organic compounds in methane (CH4). ADM1 simulates the
main biochemical (related to the microbial community) and physico-
chemical (related to the liquid or gaseous state of the variables) pro-
cesses for a reasonable modelling of several intermediate compounds
formation leading to the final production of CH4. Although the ADM1
model predicts the dynamic concentrations of biochemical components
fairly, its predictions are limited due to the insufficient consideration of
physicochemical and biochemical processes (Zhang et al., 2015a).
During the last decade, many modifications/manipulations of the

Corresponding author.
∗

E-mail addresses: luigi.frunzo@unina.it (L. Frunzo), fgfermoso@ig.csic.es (F.G. Fermoso), vincenzo.luongo@unina.it (V. Luongo),
mariarosaria.mattei@unina.it (M.R. Mattei), giovanni.esposito@unicas.it (G. Esposito).

https://doi.org/10.1016/j.jenvman.2018.11.058
Received 16 July 2018; Received in revised form 3 November 2018; Accepted 15 November 2018
Available online 22 April 2019
0301-4797/ © 2018 Elsevier Ltd. All rights reserved.
L. Frunzo, et al.
original mathematical model have been developed in order to introduce
specific process affecting the anaerobic conversion of organic sub-
strates, such as sulphur (S) degradation and kinetics (Fedorovich et al.,
2003), or CaCO3 precipitation (Batstone and Keller, 2003). ADM1 ex-
tensions have also been proposed to remove the discrepancies in both
carbon and nitrogen balances (Blumensaat and Keller, 2005), to con-
sider the effects of particle size distribution in anaerobic digestion of
complex organics (Esposito et al., 2011a), to improve the physico-
chemical ADM1 framework by incorporating more inorganic compo-
nents (metal ions and phosphates), or to include precipitation processes
(Zhang et al., 2009; Flores-Alsina et al., 2016).
Up to date, an overall model, or ADM1 extension, able to take into
account the several processes affecting TE in AD systems is missing. In
order to include TE dynamics and their effects on AD systems, ADM1
should be extended accounting for the main biological, chemical and
physical processes where TE are involved. More precisely, it is note-
worthy to include the following dynamic processes, divided in three
main categories:
Biochemical processes:
− release of metal ions, sulphur (S) and phosphorus (P) during hy-
drolysis of complex organic matter, and their uptake during biolo-
gical conversion processes;
− inhibition and stimulation effects of TE on methanogenesis reac-
tions;
Chemical processes:
− precipitation processes of metal ions taking into account the asso-
ciation/dissociation processes of carbonate, phosphate and sulphide
species;
− complexation reactions of metal and soluble components;
− metal sorption on particulate components, e.g. inert and microbial
biomasses;
Physical processes:
− liquid-gas transfer of hydrogen sulphide.
The aim of this study is to propose and apply a mechanistic model
based on ADM1 able to simulate the TE dynamics in AD systems. The
model has been synthesized to mechanistically describe the main bio-
chemical and physicochemical processes affecting TE availability and to
analyse the impact that the combination of such processes exerts on the
AD process stability and efficiency.
2. Most relevant processes affecting TE in AD systems
2.1. Input of TE in AD environment
TE in an AD environment usually come from two different sources,
i.e. TE content in the feedstock and TE supplementation as external
addition. In many cases, the release of TE from the feedstock due to the
decomposition of the organic matter and associated compounds fulfil
the micronutrient requirements for a successful process performance. A
significant example has been recently published by Lee et al. (2018),
who highlighted that the degradation of TE containing biomasses sup-
ports microbial activities during the AD process. If TE are released over
their toxic level on microorganisms, they contribute to the deterioration
of the AD performance. High TE concentrations may also limit the
proper use of digestate as fertilizer and cause environmental pollution
(Ortner et al., 2015). If TE are released below a threshold value for
microbial requirement, external addition is needed to provide the
minimum concentrations of such essential elements. This strategy is
highly recommended for AD of organic substrates with a low TE content
such as food waste and stillage (Choong et al., 2016). A wide literature
588
Journal of Environmental Management 241 (2019) 587–602
is available on the supplementation of single elements such as Fe, Ni
and Co (Fermoso et al., 2009). However, recent experimental activities
highlighted that a correct combination of TE supplementation can be
more beneficial than single element addition strategies (Moestedt et al.,
2016). According to Thanh et al. (2016), TE can be supplemented
continuously, pre-loaded or pulsed into anaerobic reactors. Moreover,
the dosing strategy of the AD systems should take into account the
operational parameters of the process, such pH, Hydraulic Retention
Time (HRT) and Organic Loading Rate (OLR), the temperature regime,
the digestion operating mode (single or double stage digestion), the
type of microbial biomass operating the process, and the reactor con-
figuration. Indeed, all these parameters can directly or indirectly in-
fluence TE speciation and thus their availability to microorganisms.
2.2. Effect of TE on biomass growth
The microbial demands for TE in anaerobic environments are as
diverse as the involved microorganisms and their functions (Wintsche
et al., 2016). Indeed, the anaerobic microorganisms have different op-
timal growth conditions with different physiologies, growth kinetics,
nutrient requirements, and sensitivity levels to environmental changes
(Mudhoo and Kumar, 2013). Several studies highlighted that TE affect
all the trophic groups due to their functions in enzyme complexes
(Wintsche et al., 2016). In AD systems, many TE, such as cobalt, nickel,
iron, zinc, molybdenum and/or tungsten, are part, in the form of central
ions conferring catalythic functions, of the essential enzymes driving
methanogenesis reactions (Zandvoort et al., 2006). In particular, Ni, Co
and Fe are essential cofactors of carbon monoxide dehydrogenase,
acetyl-CoA decarbonalyse, methyl-H4SPT:HS-CoM methyltransferase,
methyl-CoM reductase and other enzymes involved in the acetoclastic
methanogenesis pathway. In experimental studies, some TE have been
found to affect the microbial activity of propionate oxidizing bacteria as
they utilize the hydrogenase enzyme (Romero-Güiza et al., 2016). It is
assumed that methanogens groups, both hydrogenotrophic and acet-
oclastic, are usually characterized by slower growth rates compared to
the other microbial groups and could be considered as the most sensi-
tive microbial groups to TE limitations (Thanh et al., 2016). In addition,
many TE can have an inhibitory effect on the microbial metabolism
depending on the concentration level as they can deactivate enzymes by
reacting with their functional groups, denature proteins and compete
with essential cations (Zhang et al., 2009).
The stimulatory and inhibitory concentration ranges of TE on
anaerobic microorganisms are disparate; for a general overview refer to
the following recent studies (Thanh et al., 2016; Choong et al., 2016;
Romero-Güiza et al., 2016; Glass and Orphan, 2012). Such incon-
sistency in the beneficial and inhibitory levels of TE is mainly related to
the difference in the background values and TE distributions in the
feedstocks, in the diversity of the digestate texture as well as in the
characteristics of the specific microflora operating the biochemical
conversion (Cao et al., 2018). Moreover, the use of TE as AD additives,
may smooth the inhibition of the anaerobic biomass by reducing the
concentration of H2S in the biogas through precipitation processes
(Romero-Güiza et al., 2016).
2.3. Fate of sulfur and phosphorous compounds
Sulfur and Phosphorous compounds are crucial factors in AD sys-
tems as they are extremely reactive towards TE and they contextually
constitute fundamental nutrients for the anaerobic microbial consortia.
It has been reported that the dry weight of bacteria may be constituted
by 1–3% and around 1% of phosphorous and sulphur, respectively
(Gerardi, 2006). In addition, sulfur and phosphorous are involved in
various physicochemical processes, such as precipitation, affecting TE
availability and dynamics in AD systems. To account for the fate of TE
in AD systems, the understanding of sulfur and phosphorous dynamics
is a fundamental step as inorganic ligands, including phosphate and
L. Frunzo, et al.
sulphide, are assumed to control the chemical speciation of TE
(Callander and Barford, 1983a,b).
P may exist in two different forms, i.e. inorganic and organic, and
usually can be found in wastewaters in the prevalent form of ortho-
phosphate (PO43−), which represents the readily bioavailable form and
the product of poly-phosphate (H2P2O7) and organic phosphorous hy-
drolysis and mineralization (Sanchez et al., 2000, Gerardi, 2006). It is
an essential nutrient for a wide range of living organisms (De-Bashan
and Bashan, 2004; Kodera et al., 2013). It is often stated that anaerobic
conversion of organic substrates promotes the mobility and availability
of phosphorous during AD (Möller and Stinner, 2010; Bachmann et al.,
2011). Moreover, the bioaccumulation/sequestration process of phos-
phorous compounds in anaerobic environments by microbial consortia
usually competes with the precipitation process, due to the presence of
binding compounds such as TE (Keating et al., 2016; Möller and Müller,
2012). For this reason, AD has been used as an efficient strategy for
phosphorous removal leading to bioaccumulation and/or precipitation
in the form of metallic compounds, e.g. struvite crystallization, al-
lowing phosphorous recovery from digestate (De-Bashan and Bashan,
2004).
S compounds in a digester can be found in many different forms, as
in the gas, liquid and solid phase. Many industrial processes generate
high COD and high sulphur content (S-SO42-) wastewaters, which can
be used as feedstock to AD systems. Under anaerobic conditions, SRB
oxidize organic acids by using S-SO42- as final electron acceptor, thus
generating H2S and other reduced sulfur compounds (Fedorovich et al.,
2003, Möller and Müller, 2012; Yekta et al., 2014). Moreover, it has
been reported that proteins contain at least one of the sulphur amino
acids, i.e. cysteine and methionine, releasing thiol groups (-SH) during
anaerobic degradation (Gerardi, 2006). The presence of H2S, and its
anionic species HS− and S2−, affects the availability of the TE
(Gustavsson et al., 2013) and has inhibitory effects on hydro-
genotrophic methanogens (Batstone et al., 2002), acetate utilization
and, more generally, on overall microbial activities as well as methane
generation (Hilton and Oleszkiewicz, 1988; Lin, 1993).
2.4. Precipitation
Typically, AD environments are characterized by the presence of
abundant inorganic ligands, such as carbonate, phosphate and/or sul-
phide, which are assumed to control the speciation of TE by the for-
mation of poorly soluble metal precipitates (Yekta et al., 2017, Thanh
et al., 2017). Carbonate, phosphate and sulphide interact also with the
light metals, usually contained in the digestion substrates (e.g. calcium,
magnesium, potassium), leading to the formation of significant quan-
tities of struvite, hydroxyapatite and calcite. Sulphide concentration has
been recognized as a primary factor in the definition of TE bioavail-
ability as it regulates the formation of metal sulphides, which are
characterized by a very low solubility product resulting in a remarkably
low free metal ion concentration (Barrera et al., 2015; Thanh et al.,
2016). Sulphide chemistry has been claimed as the main regulator of
the trace elements bioavailability in the AD system (Fermoso et al.,
2009). In presence of sulphide, TE bioavailability has been found to
decrease in the order Fe > Ni > Co, leading to a suboptimal biogas
formation. Fe is often supplied in larger quantities than Ni and Co to
promote the precipitation of FeS and the consequent consumption of
the metabolite biosulphide produced by sulfate reducing bacteria, as
well as the adsorption and co-precipitation of Ni and Co on FeS. Indeed,
it is a common practice in full-scale anaerobic digestion to supply Fe in
the form of FeCl2 or FeCl3 in order to reduce the H2S content in the
biogas composition and thus lower the cost for H2S removal (Choong
et al., 2016). The precipitation estimation in the context of AD is largely
limited to sulfide species. Sequential extraction has largely been used to
assess the different fractions of TEs in AD systems and in some studies
the effect of precipitation/dissolution on methane production has been
explicitly investigated. For example, when metals were added
589
Journal of Environmental Management 241 (2019) 587–602
continuously at a rate of 0.05–0.2 μmol h−1, it was observed an inter-
mittent decrease in methane production (Gonzalez-Gil et al., 1999).
These rates of TE addition corresponded to TE concentrations of 1–4 μM
and are in accordance with TE requirements based on the biomass yield
and TE content of microorganisms. The authors thus concluded that
when the amount of substrate is not limiting, metal dissolution can be
the rate-limiting step. It was argued that readily dissolved precipitates
provide the required nutrients for an exponential increase in methane
production rate by the methylotrophic microorganisms. However, if the
metal uptake rate is higher than the related dissolution rate, then a TE
limited environment is created. Conversely, it was found that growth
limitation due to slow dissolution rates of sulfides is unlikely to happen
(Jansen et al., 2007). Indeed, it was found that the methanogenic ac-
tivity of Methanosarcina barkeri decreased with increase in presence of
total sulfide precipitates in the reactor. The authors argued that the
decrease in methane production may not be due to an increase in sul-
fide in the system as the consumption rates of Ni and Co did not slowed
down with increase in sulfide. Recently, it was shown by sequential
extraction that in an AD system (for slaughterhouse waste) a significant
portion of dosed TEs is not bioavailable (Ortner et al., 2015). However,
the study could not set a clear trend for carbonate bound and sulfide/
organic bound fractions in the sequential extraction experiments.
Whereas, the dissolved and exchangeable fractions increased with in-
crease in dose of TEs which also corresponds to increase methane
production and decrease in VFA accumulation.
Although mineral sulphides represent the major precipitate in
anaerobic environments at the pH range usually adopted for anaerobic
digestion (Gustavsson, 2012, Liu et al., 2015; Roussel and Carliell-
Marquet, 2016), TE precipitates of carbonate and phosphate can be
present in AD systems in significant amounts and have to be taken into
account for a correct prediction of TE bioavailability. Chemical P pre-
cipitation is based on the formation of insoluble phosphate salts (e.g.
struvite, hydroxyapatite, vivianite) and it is considered as a valuable
method for the recovery of P from digested phosphate-rich sludge
(Cheng et al., 2017). Iron can be added to promote phosphorous re-
covery in AD and vivianite crystallization has been recognized as more
competitive for ferrous ions over siderite (FeCO3) formation, which
reduces the demand of iron source to the system or omits a degassing
step for the removal of bicarbonates as an undesired ferrous sink
(Saidou et al., 2009).
2.5. Complexation
Different forms of TE are able to perform a large number of bio-
chemical processes in the liquid phase, such as formation of inorganic
and organic complexes, prior to make contact with the microbial bio-
mass (Thanh et al., 2016). Chelation of TE occurs due to their capability
of interacting with many functional groups, i.e. carboxyl, hydroxyl and
amino groups, available in AD environments. It should be noted that the
formation of soluble metal organic and inorganic complexes might have
a beneficial effect on TE bioavailability as it increases the level of total
soluble metals in an anaerobic digester competing with precipitation
kinetics (Callander and Barford, 1983a,b; Fermoso et al., 2008). The
addition of a chelating agent leads to the formation of specific TE
compounds that could be differently up-taken by the microbial biomass
depending on their nature and physical characteristics. The following
chelating agents have been investigated: nitrilotriacetic acid (NTA) (Hu
et al., 2008), ethylenediaminetetraacetic acid (EDTA) (Vintiloiu et al.,
2013), and ethylenediamine-N,N′-disuccinic acid ([S,S]-EDDS) (Zhang
et al., 2015c). In all these works, the addition of the chelating agents
resulted in an increased methane production due to the improvement in
the element bioavailability (Choong et al., 2016). However, the effect of
such chelating agents to humans and the environment must be further
assessed. Other chelating agents are represented by the soluble-micro-
bial products and extracellular polymeric substances which enhance TE
bioavailability through metal binding and complexation (Choong et al.,
L. Frunzo, et al. Journal of Environmental Management 241 (2019) 587–602

Fig. 1. Schematic representation of the conversion pathways in the mathematical model.

2016).
2.6. Sorption
Sorption is one of the main mechanisms affecting TE bioavailability.
It involves a series of processes, such as complex formation, chelation of
metals, ion exchange, adsorption, inorganic micro-precipitation and
translocation of metal into the cell, which lead to TE immobilization
and sequestration on inorganic and/or organic particulate compounds,
which reduce the bioavailability of TE for the microbial species.
Biosorption has been recognized as a promising technology for the re-
moval of several TE from contaminated wastewaters (Frunzo, 2017)
due to the high efficiency, cost effectiveness and affinity with metals,
metalloids and other pollutants from many biological materials (Gadd,
2009). Indeed, the potential of many living organisms and agricultural
wastes as biosorbents has been largely studied and reviewed (Wang and
Chen, 2009). Since most microorganisms live in the form of biofilms,
the different nature of the cell agglomerates and the presence/compo-
sition of EPS further contribute to biosorption (Flemming and
Wingender, 2010; Comte et al., 2006; Papirio et al., 2017). Weakly
acidic functional groups present in the cell wall and EPS are involved in
the mechanism proposed above. In addition to those groups, poly-
saccharides and amino acids constituting the basic structure of the cell
wall contain polarizable groups (sites), such as phosphate, carboxyl,
hydroxyl and amino-groups, which are capable of interaction with ca-
tions. These sites may contribute to the reversible TE-binding capacity
of the biomass of granular sludge (Van Hullebush et al., 2003). In
systems involving bacterial colonies forming biofilms, the higher con-
centration of EPS directly results in increasing biosorption yields. The
distribution, immobilization, and remobilization of TE depend on the
sorption properties of the biofilms, determined by many factors in-
cluding pH, redox potential, and concentration of ligands present
within the biofilm matrix (Thanh et al., 2016).
3. Mathematical model
soluble and particulate components constituting the system. The model
has been synthesized to mechanistically include the most relevant
processes affecting TE in AD systems and to analyse the impact that the
combination of such processes exerts on the AD process stability and
conversion efficiency. The processes have been implemented as sys-
tematic additions to the original ADM1. In general form, the model is
formulated in terms of four groups of state variables:
1) n1 soluble components in liquid phase Si , including the compounds
deriving from the hydrolysis of the complex organic matter, all the
forms (ionic, free and total) of the bio-products from the biochem-
ical reactions carried out by the diverse microbial groups con-
stituting the anaerobic consortium, the TE affecting the AD system
dynamics and, inorganic cations and anions;
2) n2 n1 particulate components Xi , representing the concentration of
the microbial groups carrying out the biochemical reactions, the
complex organic matter fed to the AD system, and the particulate
material deriving from the hydrolysis and disintegration steps;
3) n1 gas components Sgas, i (i.e. hydrogen, carbon dioxide, methane,
hydrogen sulphide), which are in equilibrium with the corre-
sponding components in the liquid phase;
4) m3 particulate components Xp, i , representing the mineral pre-
cipitates deriving from the mineral precipitation process involving
specific soluble compounds Si .
3.1. Mass balance equations
The differential equations governing TE, substrates, products and
bacterial groups dynamics involved in the AD processes take the fol-
lowing form:
dVliq Si
= qin Sin, i qout Si + Vliq i A, i (t , S) Vliq T , i (t , S , Sgas)
dt
m1 m
+ Vliq j = 1 i, j j
(t , S, X) Vliq j =21 S, j (t , S , X)
+ Vliq
m3 m
(t , S, Xp) Vliq j =41 C , j (t , S), i = 1, …, n1, t > 0, (1)
j = 1 i, j P , j

The proposed extended anaerobic digestion model accounting for dVliq Xi

= qin Xin, i qout Xi + Vliq
m1
j=1 i, j j (t , S , X),
TE dynamics in AD systems is based on mass conservation principles dt
i = n1 + 1, …, n2 , t > 0, (2)
and is formulated as a set of ordinary differential equations for the

590
 Table 1
Biochemical rate coefficients ( i, j ) and kinetic rate equations ( j ) for the soluble components of the modified ADM1 including TEs dynamics.

Component→ 1 2 3 4 5 6 7 8
L. Frunzo, et al.

Process ↓
Ssu Saa Sfa Sva Sbu Spro Sac SH2

1 Disintegration of OFMSW
2 Hydrolysis of RB 1
Carbohydrates
3 Hydrolysis of SB 1
Carbohydrates
4 Hydrolysis of RB Proteins 1
5 Hydrolysis of SB Proteins 1
6 Hydrolysis of RB Lipids 1-ffa,li ffa,li
7 Hydrolysis of SB Lipids 1-ffa,li ffa,li
8 Uptake of Sugars −1 (1-Ysu) fbu,su (1-Ysu) fpro,su (1-Ysu) fac,su (1-Ysu) fH2,su
9 Uptake of AminoAcids −1 (1-Yaa) fva,aa (1-Yaa) fbu,aa (1-Yaa) fpro,aa (1-Yaa) fac,aa (1-Yaa) fH2,aa
10 Uptake of LCFA −1 (1-Yfa) 0.7 (1-Yfa) 0.3
11 Uptake of Valerate −1 (1-YC4) 0.54 (1-YC4) 0.31 (1-YC4) 0.15
12 Uptake of Butyrate −1 (1-YC4) 0.8 (1-YC4) 0.2
13 Uptake of Propionate −1 (1-Ypro) 0.57 (1-Ypro) 0.43
14 Uptake of Propionate by −1 (1-YpSRB) 0.57
pSRB
15 Uptake of Acetate −1
16 Uptake of Acetate by aSRB −1
17 Uptake of Hydrogen −1
18 Uptake of Hydrogen by −1
hSRB
19 Decay of Xsu

591
20 Decay of Xaa
21 Decay of Xfa
22 Decay of XC4
23 Decay of Xpro
24 Decay of XpSRB
25 Decay of Xac
26 Decay of XaSRB
27 Decay of XH2
28 Decay of XhSRB
Monosaccarides Amino Acids Long Chain Fatty Acids Total Valerate Total Butyrate Total Propionate Total Acetate Liquid Hydrogen
(kg COD m−3) (kg COD m−3) (kg COD m−3) (kg COD m−3) (kg COD m−3) (kg COD m−3) (kg COD m−3) (kg COD m−3)

9 10 11 12 13 14 15 16 Rate rj
−3−1
[kgCODmd /
−3−1
SCH4 SIC SIN SIP SIS SSO SI SMe kmol md ]

1 fSI,Xc 1
2 2
3 3
4 f_IS f_Maa 4
5 f_IS f_Maa 5
6 f_IP 6
7 f_IP 7
8 i = 1 9,11 16,38 55 Ci i,8
-(Ysu)Nbac

9 i = 1 9,11 16,38 55 Ci i,9

Naa-(Ysu)Nbac

(continued on next page)

Journal of Environmental Management 241 (2019) 587–602
 Table 1 (continued)

9 10 11 12 13 14 15 16 Rate rj
−3−1
[kgCODmd /
L. Frunzo, et al.

−3−1
SCH4 SIC SIN SIP SIS SSO SI SMe kmol md ]

10 -(Yfa)Nbac 10
11 -(YC4)Nbac 11
12 -(YC4)Nbac 12
13 i = 1 9,11 16,38 55 Ci i,13
-(Ypro)Nbac

14 i = 1 9,11 16,38 55 Ci i,14

-(YpSRB)Nbac (1-YpSRB)0.43/64 -(1-YpSRB)0.43/64

15 (1-Yac) i = 1 9,11 16,38 55 Ci i,15

-(Yac)Nbac
-(Yac)Mbac
16 i = 1 9,11 16,38 55 Ci i,16
-(YaSRB)Nbac (1-YaSRB)/64 -(1-YaSRB)/64
-(YaSRB)Mbac
17 (1-Yac) i = 1 9,11 16,38 55 Ci i,17
-(YH2)Nbac
-(YH2)Mbac
18 i = 1 9,11 16,38 55 Ci i,18
-(YhSRB)Nbac (1-YhSRB)/64 -(1-YhSRB)/64
-(YhSRB)Mbac
19 i = 1 9,11 16,38 55 Ci i,19 i = 11,39 54 Ni i,19

20 i = 1 9,11 16,38 55 Ci i,20 i = 11,39 54 Ni i,20

21 i = 1 9,11 16,38 55 Ci i,21 i = 11,39 54 Ni i,21

592
22 i = 1 9,11 16,38 55 Ci i,22 i = 11,39 54 Ni i,22

23 i = 1 9,11 16,38 55 Ci i,23 i = 11,39 54 Ni i,23

24 i = 1 9,11 16,38 55 Ci i,24 i = 11,39 54 Ni i,24

25 i = 1 9,11 16,38 55 Ci i,25 i = 11,39 54 Ni i,25

26 i = 1 9,11 16,38 55 Ci i,26 i = 11,39 54 Ni i,26

27 i = 1 9,11 16,38 55 Ci i,27 i = 11,39 54 Ni i,27

28 i = 1 9,11 16,38 55 Ci i,28 i = 11,39 54 Ni i,28

Liquid Methane Inorganic Carbon Inorganic Nitrogen Inorganic Phosphorous Total sulphides Total sulphates Soluble Inerts Soluble Metal
(kg COD m−3) (kmol C m−3) (kmol N m−3) (kmol P m−3) (kmol S m−3) (kmol S m−3) (kg COD m−3) (kmol Me m−3)
Journal of Environmental Management 241 (2019) 587–602
L. Frunzo, et al. Journal of Environmental Management 241 (2019) 587–602

dVgas Sgas, i
= qgas Sgas, i + Vliq reaction rates adopted in the proposed model will be detailed in the
T , i (t , S, Sgas),
dt
following sections.
i = 1, …, n1, t > 0, (3)
dVliq Xp, i 3.2. Biochemical reaction rates
dt
= qin X pin, i qout Xp, i + Vliq P , i (t , S, Xp),
i = 1, …, m3, t > 0, (4) The large applicability and high versatility of the original ADM1 is
where: due to the simple structure of the mathematical model and the rea-
sonable prediction of the main products deriving from the anaerobic
n1 denotes the number of soluble components; conversion of complex organic matter. Several extensions have been
n2 n1 denotes the number of particulate components; proposed over the last decade to specifically focus on relevant processes
m1 denotes the number of biochemical processes taken into account; and biological interactions occurring in AD due to specific substrate
m2 denotes the number of sorption processes taken into account; compositions or reactor configurations. In agreement with the sche-
m3 denotes the number of precipitation processes taken into ac- matic approach of ADM1, five groups of biochemical reactions have
count; been individuated to account for TE dynamics in AD (Fig. 1): i) the
m4 denotes the number of complexation processes taken into ac- disintegration of complex organic compounds XC in carbohydrates Xch ,
count; proteins Xp , lipids Xli and, particulate inerts XI , ii) the hydrolysis of
these compounds in monosaccharides Xsu , amino acids Xaa , and fatty
i, j is the stoichiometric coefficient of species i on biochemical
process j; acids Xfa and XC4 , and the contextual release of inorganic carbon XIC ,
nitrogen XIN , phosphorous XIP , sulfur XIS and TEs SMe2 + , iii) the acid-
i, j is the stoichiometric coefficient of species i on precipitation
process j; ogenesis, iv) the acetogenesis and v) the methanogenesis steps.
The disintegration step has been modelled by using a surface based
i is the stoichiometric coefficient for the acid base reaction invol-
ving the ith soluble component; kinetic approach Eq. (10) which explicitly takes into account the spe-
Si denotes the ith soluble component, S = (S1, …, Sn1) ; cific surface area of organic compounds (Esposito et al., 2011b). The
Xi denotes the ith particulate component, X = (Xn1+ 1, …, Xn2 ) ; biodegradability characteristics of the hydrolysable substrates (carbo-
Sgas, i denotes the ith component in gas form, Sgas = (Sgas,1, …, Sgas, n1) ; hydrates, proteins and lipids) have been taken into account by the
Xp, i denotes the ith precipitate, Xp = (Xp,1, …, Xp, m3 ) ; addition of supplementary state variables accounting for the rapid
j (t , S , X) represents the rate of the jth biochemical process;
( Xch R , Xpr R and Xli R ) or slow ( Xch S , Xpr S and Xli S ) biodegrad-
S , j (t , S , X) represents the rate of the jth sorption process;
ability of these compounds (Esposito et al., 2011a,b). First order ki-
netics have been used to describe the hydrolysis of these compounds in
P , j (t , S , X p) represents the rate of the jth precipitation process;
anaerobic environments (Eqs. (11)–(16)).
C , j (t , S) represents the rate of the jth complexation process;
According to (De Gracia et al., 2006; Flores-Alsina et al., 2016), the
A, i (t , S) represents the acid base kinetic rate equation for the ith
soluble component; fate of sulphur and phosphorous have been included into the original
model by the addition of new state variables, namely total sulphates
T , i (t , S , S gas) represents the gas transfer rate for the ith soluble
component. SSO , total sulphides SIS and total phosphorous SIP , as reported in the
modified biochemical matrix of the proposed model (Tables 1 and 2).
Note that although the total amount of TE in anaerobic effluents is The oxidized sulphur is usually supplemented to the system through
normally used as a practical index of TE bioavailability, the effective industrial wastewaters. The last two compounds instead originate from
biouptakable amount of TE mainly depends on the concentrations of the the hydrolysis of proteins and amino acids (Shi and Chance, 2011) and
free metal ions within the reactor. For this reason, new dynamic state then assume different forms according to the acid base reactions re-
variables, which account for the concentration of the generic free metal ported in the following section. Similarly, TE are supposed to be sup-
ion in liquid phase, have been introduced for the mathematical mod- plemented to the anaerobic system or contained in the digestion sub-
elling of all the biological and physicochemical phenomena affecting TE strates. The supplementation is modelled as an inlet flux to the reactor
bioavailability in anaerobic systems. Moreover, a charge balance ac- and it is expressed as a function of the inlet flow rate and the con-
counting for all the ionic species is needed to completely define the centration of the supplemented metal ion. The TE contained in the
model as the pH of the system represents a major biological inhibition feedstock are supposed to be included mainly in the protein fraction of
factor. Such equation can be written in general form as follows: the digestion substrate. The main reason relies on the predominance of
p q
metal ions within the functional groups characterizing these com-
Qi+ Qi = 0, p + q < n1, pounds. TE release into the liquid phase derives from the hydrolysis of
i=1 i=1 (5) proteins and the amino acids bioconversion. Therefore, a specific pro-
duction rate has been considered in the mass balance equation related
where
to the generic metal ion. According to other studies (Fedorovich et al.,
p defines the number of cationic components;
2003; Batstone, 2006; Barrera et al., 2015), sulphate reduction has been
q defines the number of anionic components;
incorporated in the original ADM1 including the inhibitory effect of the
Qi+ represents the cationic equivalent concentration of species ith;
produced sulphide on methanogenic groups causing potential toxicity
Qi represents the anionic equivalent concentration of species ith.
and corrosion (Flores-Alsina et al., 2016). The behaviour of sulphate
The differential algebraic system obtained by coupling Eq. (3.5) to
reducing bacteria (SRB) competing for different intermediates, i.e.
the mass balance equations (3.1)–(3.4) can be solved after introducing
propionate, acetate and hydrogen, has been included by adding the new
the following initial conditions:
state variables XpSRB , XaSRB and XhSRB (Barrera et al., 2015). According
Si (0) = Si0, i = 1, …, n1, (6) to Fedorovich et al. (2003), dual term Monod type kinetics have been
used to describe the reduction of SSO by SRB using the above-mentioned
Xi (0) = Xi0 , i = n1 + 1, …, n2, (7) electron donors. In addition, the biomass decay processes related to SRB
0 have been included in the model and described as first order kinetics.
Sgas, i (0) = Sgas , i, i = 1, …, n1, (8)
To cover the discrepancy between the nitrogen content in the biomasses
Xp, i (0) = X p0, i , and the feeding composite material in the original ADM1, additional
i = 1, …, m3. (9)
terms related to the decay processes have been included for the in-
The biochemical, acid/base, gas transfer, sorption, precipitation organic nitrogen as in Blumensaat and Keller (2005). The inhibition

593
 Table 2
Biochemical rate coefficients ( i, j ) and kinetic rate equations ( j ) for the particulate components of the modified ADM1 including TEs dynamics.

Component→ 38 39 40 41 42 43 44 45 46
L. Frunzo, et al.

Process ↓
XC Xch_R Xch_S Xpr_R Xpr_S Xli_R Xli_S Xsu Xaa

1 Disintegration of OFMSW −1 fCh,Xc fr fCh,Xc fs fPr,Xc fr fPr,Xc fs fLi,Xc fr fLi,Xc fs

2 Hydrolysis of RB −1
Carbohydrates
3 Hydrolysis of SB −1
Carbohydrates
4 Hydrolysis of RB Proteins −1
5 Hydrolysis of SB Proteins −1
6 Hydrolysis of RB Lipids −1
7 Hydrolysis of SB Lipids −1
8 Uptake of Sugars Ysu
9 Uptake of AminoAcids Yaa
10 Uptake of LCFA
11 Uptake of Valerate
12 Uptake of Butyrate
13 Uptake of Propionate
14 Uptake of Propionate by pSRB
15 Uptake of Acetate
16 Uptake of Acetate by aSRB
17 Uptake of Hydrogen
18 Uptake of Hydrogen by hSRB
19 Decay of Xsu 1 −1
20 Decay of Xaa 1 −1
21 Decay of Xfa 1

594
22 Decay of XC4 1
23 Decay of Xpro 1
24 Decay of XpSRB 1
25 Decay of Xac 1
26 Decay of XaSRB 1
27 Decay of XH2 1
28 Decay of XhSRB 1
Organic Composites R.Biodegradable S.Biodegradable R.Biodegradable S.Biodegradable R.Biodegradable S.Biodegradable Sugars Amino Acids
(kg COD m−3) Carbohydrates Carbohydrates Proteins Proteins Lipids Lipids Degraders Degraders
(kg COD m−3) (kg COD m−3) (kg COD m−3) (kg COD m−3) (kg COD m−3) (kg COD m−3) (kg COD m−3) (kg COD m−3)

47 48 49 50 51 52 53 54 55 Rate rj
−3
[kgCODm-
−1
Xfa XC4 Xpro XpSRB Xac XaSRB Xh2 XhSRB XI d /
−3−1
kmol md ]

1 fXi,Xc 1
2 2
3 3
4 4
5 5
6 6
7 7
8 8
9 9
10 Yfa 10
11 YC4 11
12 YC4 12
Journal of Environmental Management 241 (2019) 587–602
L. Frunzo, et al. Journal of Environmental Management 241 (2019) 587–602

terms for the uptake rates of the diverse microbial groups have been

−3−1
]
adopted from Barrera et al. (2015). TE have been proposed to affect
−3
[kgCODm-

kmol md
Rate rj

/
mainly the methanogenic activity, both hydrogenotrophic and acet-
−1

21
20
14

17

24
13

15
16

18
19

22
23

25
26
27
28
oclastic as methanogens have been proved to be strongly affected by TE
d

deprivation (Wintsche et al., 2016). Based on these reasonings, the

microbial kinetics of hydrogenotrophic and acetoclastic methanogens

Particulate Inerts
have been modified by including an inhibition function which depends

(kg COD m−3)

on the concentration of the free metal ions in the liquid phase (IMe2 +).
Such function accounts for the inhibition of methanogenic activity due
to TE deprivation and it has been modelled as a secondary substrate
55

XI

inhibition term. The functional forms of the inhibition terms Ii are re-
ported in Table 3. Furthermore, a consumption term has been in-
troduced in the mass balance for the generic free metal ion in liquid
phase to take into account the microorganisms' uptake of TEs. All the

(kg COD m−3)

SRB Hydrogen
kinetic rate equations i have been following listed according to the

Degraders
ADM1 and its further extensions (Esposito et al., 2011b; Frunzo et al.,
XhSRB

YhSRB

2012; Barrera et al., 2015):

−1
54

1 = Ksbk Ca , (10)
Hydrogen Degraders
(kg COD m−3) 2 = Khyd _ ch _ R X ch _ R , (11)

3 = Khyd _ ch _ S Xch _ S , (12)

4 = Khyd _ pr _ R Xpr _ R , (13)

−1
Xh2

Yh2
53

5 = Khyd _ pr _ S Xpr _ S , (14)

SRB Acetate Degraders

6 = Khyd _ li _ R Xli _ R , (15)

(kg COD m−3)

7 = Khyd _ li _ S Xli _ S , (16)

Ssu
XaSRB

YaSRB

= Km, su Xsu I1,

−1

8
(17)
52

K ssu + Ssu

Saa
9 = Km, aa Xaa I1,
(18)
Acetate Degraders

K saa + Saa
(kg COD m−3)

Sfa
10 = Km, fa Xfa I2,
K sfa + Sfa (19)
−1
Xac

Yac
51

Sva 1
11 = Km, c 4 XC 4 I3,
K sc4 + Sva 1 + Sbu /Sva (20)

Sbu 1
SRB Propionate

(kg COD m−3)

12 = Km, C 4 XC 4 I3,
K sc4 + Sbu 1 + Sva/ Sbu (21)
Degraders

Spro
XpSRB

YpSRB

= Km, pro Xpro I3,

−1

13
50

K spro + Spro (22)

Propionate Degraders

Spro SSO
14 = Km, pSRB XpSRB I6,
K spro + Spro K sSO4 pSRB + SSO (23)
(kg COD m−3)

Sac
15 = Km, ac Xac I4 ,
K sac + Sac (24)
Xpro

Ypro

−1
49

Sac SSO
16 = Km, aSRB XaSRB I6,
K sac + Sac K sSO4 aSRB + SSO (25)
Valerate and Butyrate

Sh2
(kg COD m−3)

17 = Km, h2 Xh2 I5,

K sh2 + Sh2 (26)
Degraders

Sh2 SSO
= Km, hSRB XhSRB I6,
−1
XC4

18
48

K sh2 + Sh2 K sSO4 hSRB + SSO (27)

(kg COD m−3)

= K dec, Xsu Xsu , (28)

Table 2 (continued)

19
Long Chain
Fatty Acids
Degraders

20 = K dec, Xaa Xaa , (29)

−1
Xfa
47

21 = K dec, Xfa Xfa , (30)

13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28

22 = K dec, Xc4 X c 4 , (31)

595
L. Frunzo, et al. Journal of Environmental Management 241 (2019) 587–602

Table 3
Functional form of the inhibition terms adopted in the proposed model.

Inhibition Term Expression Used for Reference

IpH Uptake of sugars, amino acids, LCFA, VFAs, hydrogen and sulphate reduction. Batstone et al. (2002)
( )
2
pH pHUL
exp 3 , pH < pHUL
pHUL pHLL

1, pH > pHUL
IIN , lim 1 Uptake of sugars, amino acids, LCFA, VFAs and hydrogen. Batstone et al. (2002)
1 + K S , IN / SIN
IH2 1 Uptake of LCFA and VFAs. Batstone et al. (2002)
1 + S H2 / KI , H2
IH2 S 1 Uptake of VFAs, hydrogen and sulphate reduction. Barrera et al. (2015)
1 + S H2 S / KI , H2 S
INH3 1 Uptake of acetate. Batstone et al. (2002)
1 + SNH3 / KI , NH3
IMe2 + 1 Uptake of acetate and hydrogen. This Study
1+K /S
I , Me2 + Me2 +

23 = K dec, Xpro Xpro , (32) H2 S H+ + HS , pka = 7.05, (46)

24 = K dec, XpSRB XpSRB , (33) HS H+ + S 2 , pka = 14, (47)

25 = K dec, Xac Xac , (34) H2 PO4 H+ + HPO42 , pka = 7.21, (48)

26 = K dec , XaSRB XaSRB , (35) HPO42 H+ + PO43 , pka = 12.32. (49)

27 = K dec, Xh2 Xh2 , (36) The rate coefficients and the kinetic rate equations for all the acid
base reactions implemented in the proposed model have been for-
28 = K dec, XhSRB XhSRB . (37) mulated as suggested by Rosen and Jeppsson (2006) and are reported in
Table 4. Note that for each acid base pair Si/ Si + 1, where Si corresponds
The inhibition terms in Eqs. (17)–(27) have been modified from
to the acidic component, the kinetic rate expressions are related as
Barrera et al. (2015) to consider the effect of TE on the microbial ki-
follows:
netics. They are formulated as follows:
A, i + 1 = A, i , (50)
I1 = IpH · IIN , lim, (38)
= KA / Bva (Sva (S H + + K a, va) Sva) , (51)
I2 = IpH · IIN , lim·IH2, (39) A,17

I3 = IpH · IIN , lim·IH2 ·IH2 S , (40) A,19 = KA / Bbu (Sbu (S H + + K a, bu) Sbu ), (52)

I4 = IpH · IIN , lim·INH3·IH2 S · IMe2 +, (41) A,21 = KA / Bpro (Spro (S H + + K a,pro) Spro), (53)
I5 = IpH ·IIN , lim· IH2 S ·IMe2 +, (42) A,23 = KA / Bac (Sac (S H + + Ka,ac) Sac ) , (54)
I6 = IpH ·IH2 S . (43) = KA / Bco2 (Shco3
A,25 (S H + + K a,co2) + K a, co2 Sco32 SIC ), (55)

3.3. Acid-base process rate A,26 = KA / Bhco3 (Sco32 (S H + + Ka,hco3) + K a,hco3 Sco2 SIC ) , (56)

= KA / Bnh4 (Snh3 (S H + + K a, nh3) Snh4), (57)

Additional acid-base reactions have been implemented in the pro- A,28

posed model in order to take into account specific ionic forms of the
= KA / h2po4 (Shpo42 (S H + + K a,h2po4) + K a,h2po4 Spo43 SIP ) , (58)
soluble components, reacting with the bioavailable forms of TE in A,30

physicochemical processes such as precipitation and complexation. For = KA / hpo4 (Spo43

what concerns inorganic carbon, the acid-base pair HCO3 /CO32 has A,31 (S H + + Ka,hpo4) + K a,hpo4 Sh2po4 SIP ) , (59)
been introduced according to Eq. (44), due to the relevant percentage of = KA / h2s (Shs
A,33 (S H + + K a, h2s) + K a,h2s S s2 SIS ) , (60)
carbonate precipitates usually found in AD effluents (Langerak et al.,
1999). According to Barrera et al. (2015), the acid-base reaction in- = KA / hs (S s2 (S H + + K a,hs) + K a,hs Sh2s SIS ) ,
A,34 (61)
volving the undissociated sulphuric acid H2 SO4 has been neglected as it
can be considered completely dissociated in water (pKa < −2). To A,36 = KA / Bso (Sso42 (S H + + Ka,hso4) Sso4 ) , (62)
account for SRB metabolism and sulphur rich wastewaters, the che-
mical equilibrium related to the pair HSO4 / SO42 has been included
according to Eq. (45). In relation to reduced sulphur compounds, the 3.4. Gas-transfer process rate
acid-base pairs H2 S /HS and HS / S 2 have been considered according
to Eqs. (46) and (47). The sulphide ions S 2 have been introduced to The liquid-gas transfer processes for the variables
effectively model the formation of sulphide precipitates. Similarly, the SH 2, SCH 4, SIC , SIS in the liquid phase have been considered. The for-
acid-base reactions accounting for the equilibrium pairs H2 PO4 /HPO42 mulation of the kinetic rate expressions follows the approach of Rosen
and HPO42 / PO43 have been implemented according to Eqs. (48) and and Jeppsson (2006) for SH 2, SCH 4, SIC and Barrera et al. (2015) for SIS
(49). The dissociation of H3 PO4 has not been considered due to the low (see Table 5). For all the other components in liquid phase T , i has been
pKa characterizing such reaction (pKa = 2.14). considered equal to zero.

HCO3 H+ + CO32 , pka = 10.33, (44) T ,8 = KLa (Sh2 16KH , h2 pgas, h2 ), (63)

HSO4 H+ + SO42 , pka = 1.99, (45) T ,9 = KLa (Sch4 64KH , ch4 pgas, ch4 ), (64)

596
L. Frunzo, et al. Journal of Environmental Management 241 (2019) 587–602

Table 5
Liquid phase yield coefficients and rate equations ( T ,i ) for the liquid-gas
[kmol m−3d−1] transfer processes implemented in the proposed model.

Component→ 8 9 10 13 Rate rj
Process ↓ [kgCODm−3d−1]
Rate rj

SH2 SCH4 SIC SIS

A,17
A,19
A,21
A,23
A,25
A,26
A,28
A,30
A,31
A,33
A,34
A,36
T8 H2 Transfer −1 T,8
2-

T9 CH4 Transfer −1
SSO4

−1
T,9
37

T10 CO2 Transfer −1 T,10

T12 H2S Transfer −1
−

T,12
SHSO4
36

1
for the acid base reactions in the differential implementation of the modified ADM1 including TEs dynamics.

T ,10 = KLa (Sco2 KH , co2 pgas, co2 ), (65)

2-

−1
35

SS

= KLa (Sh2s 48KH , h2s pgas, h2s ), (66)

−

T ,12
−1
SHS
34

1
SH2S

3.5. Sorption process rate

33

1
3-

The sorption processes have been implemented in the proposed

SPO4

−1
32

model by following the approach introduced in D'Acunto et al. (2015)

and D'Acunto et al. (2018). A non-reversible mechanism for TE sorption
SHPO42-

has been considered. The generic TE SMe2 + has been supposed to adsorb
−1
31

on the various particulate components, including the microbial bio-

masses constituting the anaerobic consortium and the inert biomass.
SH2PO4−

Each of these components is characterized by a specific concentration of

30

binding sites, which can be found in two states: occupied or free. The
1

fraction of free binding sites evolves over time due to the production of
Snh3

−1
29

new biomass and the progressive sorption of TE. Conversely, the frac-
tion of occupied binding sites can only increase over time as the sorp-
+
Snh4

tion phenomenon has been modelled as a non-reversible process. The

28

rate characterizing the jth sorption process has been formulated as

2-

follows:
Sco3

−1
27

¯X
S, j = K s, Xn1+ 1 + j SMe2 + (1 n1+ 1 + j ) N Xn1+ 1 + j Xn1+ 1 + j , j = 1, …, m2 , (67)
Shco3-

−1
26

where
1

Ks, Xn1+1 + j represents the sorption constant on the particulate com-

SCO2
25

ponent Xn1+ 1+ j ;
1

n1+ 1 + j denotes the fraction of occupied binding sites on the com-

¯X
−1
-
Sac
24

ponent Xn1+ 1+ j ;
N Xn1+ 1 +j is the site density of the component Xn1+ 1+ j .
Shac
23

Note that the sorption process on the complex organic matter

1

XC = Xn1+ 1 has not been included in the proposed model. The number
Spro-

−1
22

of sorption processes implemented in the present model coincides with

the number of particulate components (except the complex organic
Shpro

matter) taken into account in the biochemical reactions

21

1
A, i )

m2 = n2 (n1 + 1) = 17 .
Sbu-

−1
Biochemical rate coefficients ( i ) and kinetic rate equations (

20

3.6. Precipitation process rate

Shbu
19

Diverse precipitation processes affect TE availability in AD systems

−1
-
Sva
18

(Zhang et al., 2015a). The precipitation is supposed to occur when the

solution is oversaturated for the soluble components involved in the
Shva
17

specific precipitation process. It depends on the specific solubility

product (Ksp), representing the maximum number of undissociated ions
Dihydrogen sulphide acid base
Hydrogen Phosphate acid base
Inorganic nitrogen acid base

remaining in equilibrium with the undissolved phase. According to

Carbon dioxide acid base

Zhang et al. (2015a), the precipitation rates have been formulated as

Bicarbonate acid base
Propionate acid-base

kinetic based rate equations, whose expression is reported as in the

Phosphate acid base

Sulphide acid base

Sulphate acid base
Butyrate acid-base
Valerate acid-base

Acetate acid base

following equation:
Component→

1 v
Process ↓

v
+ +
1
prec = kr , m + A v
v
[([M m ]v ) [Aa ]v ] v ksp, m ,Av
v+

(68)
Table 4

A17
A19
A21
A23
A25
A26
A28
A31
A32
A34
A35
A37

where:

597
L. Frunzo, et al. Journal of Environmental Management 241 (2019) 587–602

Table 6
Precipitation rate coefficients ( i, j ) and rate equations ( P, j ) for the precipitation processes implemented in the proposed model.

Component→ 8 10 12 13 P,1 P,2 P,3 Rate P, j

Process ↓ [kmol m−3d−1]
2+
SMe SIC SIP SIS XMeCO3 XMe3(PO4)2 XMeS

P1 Carbonate precipitation −1 −1 1 P,1

P2 Phosphate precipitation −3 −2 1 P,2
P3 Sulphide precipitation −1 −1 1 P,3

pH 10-3 Sulfide Speciation

9 1
S 2-
8 H 2S

[Mol L -1]
HS -
pH

7 0.5

5 0
0 20 40 60 80 100 120 0 20 40 60 80 100 120
Time [d] Time [d]
Cumulative biogas production 10-5 Iron
1.5 4
CH 4gas freeFe
CH 4 [LL -1d -1]

[Mol L -1]

1 3

0.5 2

0 1
0 20 40 60 80 100 120 0 20 40 60 80 100 120
Time [d] Time [d]
10-8 IronSulfide Pr & AA
3 8
FeS Pr
S 2- 6 AA
[gCOD L -1]
[Mol L ]

2
-1

4
1
2

0 0
0 20 40 60 80 100 120 0 20 40 60 80 100 120
Time [d] Time [d]
Fig. 2. Experimental values (square) and model predictions (continuous lines).

m v+ A v Concentrations of cations and anions coefficients related to the precipitation processes implemented in this
kr , m + A v is the precipitation rate constant model are reported in Table 6.
v
ksp, m +, A v is the solubility product 1/2 2
v
P ,1 = KP,1 ((SMe2 + SCO32 )1/2 kspk ,1 ) , (69)
v+ and v are the total number of cationic and anionic species and
v = v+ + v . 3 2 1/5 1/5 2
= KP,2 ((SMe 2+ S 3 ) kspk ,2 ) ,
P ,2 PO 4 (70)
For each precipitating compound, the free cations and anions, ty-
1/2 2
pically inorganic carbon, inorganic sulphur and inorganic phosphorous, P ,3 = KP,3 ((SMe2 + S S2 )1/2 kspk ,3 ) . (71)
dynamically react following specific kinetics, which are regulated by
the charge balance and the precipitation and solubility constant,
namely kr , m + A v .and ksp, m +, A v , respectively. In the proposed model, 4. Application and numerical results
v v
three precipitation rates have been included for the generic trace ele-
ment SMe2 + . The latter is supposed to precipitate in the form of carbo- FW is one of the most promising waste organic substrates to be used
nate, sulphide and phosphate according to Eqs. (69)–(71). The rate as feedstock for AD and renewable energy production. It has been es-
timated that FW production in China could approximately reach

598
L. Frunzo, et al.
Table 7
Operational parameters used for the numerical simulations.
Parameter Unit
Digester volume mL
Headspace volume mL
Influent FW rate mL d−1
Total FW Carbohydrates fraction – 0.65
Total FW Proteins fraction – 0.20
Total FW Lipids fraction – 0.05
Total FW Inert fraction – 0.10
Temperature °C
Sulfur content of Proteins mol gCOD−1
Sulfur content of Lipids mol gCOD−1
Iron content of AA mol gCOD−1
Iron content of Lipid mol gCOD−1
Iron content of inoculum mol gCOD−1
Iron supply mol L−1
Henry's law coefficient for H2S mol bar−1
Gas-Liquid H2S transfer coefficient d−1
FeS solubility product (Pr_FeS) mol L−1
FeS precipitation rate constant (Ksp_FeS) d−1
Stimulation constant of Fe2+ (KI,Fe) mol L−1
1.4 × 108 tons per year in 2020, which is equivalent to 10 million tons
of coal based on energy conversion by electricity production (Ye et al.,
2018). Despite the advantages deriving from the conversion of carbon-
rich substrate via AD, nowadays the key issues of FW bio-conversion lie
on the instability and unefficiency of AD reactors in long-term appli-
cations (Zhang and Jahng, 2012). This is mainly due to the lack of some
nutrients (i.e. TE) that are limiting for the anaerobic microorganisms
metabolism and induce VFA accumulation and poor process perfor-
mance for FW anaerobic conversion (Romero-Güiza et al., 2016). To
overcome this problem, co-digestion with TE-rich substrates (such as
piggery wastewater) and TE external supplementation have been
identified as promising strategies for the optimization of FW anaerobic
conversion (Zhang et al., 2015b, Romero-Güiza et al., 2016).
The availability of a mathematical model able to predict TE defi-
ciency and the optimal supplementation strategy is a fundamental re-
quirement for the management of AD bio-reactors at full scale. For this
reason, the proposed mathematical model has been applied to two il-
lustrative cases: the first one is based on the experimental results
achieved by Zhang and Jahng (2012), who examine the effect of trace
metals limitation in an AD process of FW. The second case investigates
the effects of different iron dosing on methane production and it is
aimed at individuating the optimal external iron supplementation in
long-term operations. In both cases, iron has been considered as the
only limiting TE due to its recognized high requirements in AD systems
(Choong et al., 2016).
Application 1: Effect of iron limitation in an AD process of FW.
The first numerical application has been based on the experimental
results of Zhang and Jahng (2012). In their work, the authors carried-
out different semi-continuous AD experiments for FW conversion aimed
at identifying the role of the addition of an individual trace element or a
solution containing different TE in AD bio-reactors.
To this aim, seven reactors were operated in parallel. Six reactors
were supplied with individual (or combined) TE solutions while one
rector was operated for 95 days without any trace element addition. By
analysing the behaviours of the six reactors, the authors highlighted the
remarkable effect of the low iron content in the FW that differently
from Co, Mo and Ni, plays a crucial role for the stability of the process.
The experiment related to the reactor without TE addition (cfr.
Reactor R1) has been qualitatively reproduced in silico (Fig. 2). The
operational parameters used for the numerical simulations have been
reported in Table 7.
The values of the kinetic and stoichiometric parameters as well as
the Henry's constants have been selected according to the suggested
599
Journal of Environmental Management 241 (2019) 587–602
Simulation Set Simulation Set 2
1
150 150
30 30
5 5
0.65
0.20
0.05
0.10
37 37
0.075 0.0075
0.015 0.015
0.0001 0.0001
0.00075 0.00075
0.255 0
0 5*10−3-1*10−7
0.1 0.1
200 200
104 104
18 18
0.0724 0.0724
parameter value for mesophilic AD (Batstone et al., 2002). In addition,
the kinetic disintegration constant Ksbk and the hydrolysis kinetic
constants for all simulations have been derived from literature (Esposito
et al., 2012). The organic substrate has been fractionated as 50%
readily biodegradable and 50% slowly biodegradable. The kLa coeffi-
cients for liquid-to-gas transfer of methane and hydrogen have been
adopted according to Esposito et al. (2011a) and (2011b), respectively,
whereas the kLa value of 200 d−1 (Barrera et al., 2015) has been applied
for hydrogen sulfide.
In accordance to the experimental evidence, iron represents the
limiting factor leading to the reactor failure after 80 days of operation.
To properly model the effect of iron deprivation on AD process, a
specific inhibition term has been introduced as reported in Table 3. The
expression for IFe2 + has been calibrated by setting a kinetic constant
KI , Fe2 + to the value of 0.0724 mol L−1.
The model outputs have been reported in Fig. 2. Numerical simu-
lations demonstrated the model capability of predicting pH, sulphide
speciation, methane production, free iron, iron sulphide, protein and
amino acid concentrations. Notably, the pH evolution over time and the
methane production predicted by the model have been compared with
the experimental data. The model reproduces in a reasonable way both
the methane and pH profiles and it was able to accurately predict the
process failure after about 80 operation days.
Due to the semi-continuous operation, the iron concentration de-
creases and the low iron content of the FW gradually determines the
failure of the reactor. Indeed, the inhibition of methanogenesis results
in VFA accumulation and fast decrease of the reactor pH. The pH value
drops from 7.5 to 5; the sulphide speciation changes and the amino acid
hydrolysis efficiency decreases. Indeed, when the pH reaches the value
of 5, the sulphide is mainly present in the form of H2S, which leads to
the increase of H2S gas production (data not shown) and the consequent
inhibition of amino acid hydrolysis. The latter causes an additional
decrease of free iron content into the reactor.
Application 2: Effect of iron supply in an AD process of FW.
Figs. 3 and 4 show the results of model simulations (Simulation Set
2, Table 7) performed to assess the effect of different external iron
supply on AD performance. This model application accounts for the
effect of increasing free iron concentration on sulfide speciation and
biogas composition during the biological anaerobic conversion of FW.
The external addition of iron in the range of
0.005–0.000005 mol L−1 has been investigated to explore the dynamic
effects of iron addition and optimize the TE dosing strategy. To reach a
stable initial condition (day 0), 100 simulation days have been
L. Frunzo, et al. Journal of Environmental Management 241 (2019) 587–602

CH 4 production Free Iron concentration

2 10-2

1.5

Fe2+ [Mol L -1]

CH 4 [LL -1d -1]

10-3

10-4
0.5

Increasing Iron supply

0 10-5
-100 0 100 200 -100 0 100 200
Time [d] Time [d]

H 2S production
10-4 10-6 Iron Sulfide concentration
5 3
Increasing Iron supply
2.5
4
H 2S [LL -1 d-1]

FeS [Mol L -1]

2
3
1.5
2
1

1
0.5
Increasing Iron supply
0 0
-100 0 100 200 -100 0 100 200
Time [d] Time [d]
0.005M Fe In 0.0005M Fe In 0.00005M Fe In 0.000005M Fe In

Fig. 3. Methane production, free iron concentration, hydrogen sulphide gas production and iron sulphide concentration with increasing iron supplementation.

considered for all the simulation sets.
As it is possible to notice (Fig. 3), in all the experiments the methane
production profile denotes the same trend as increasing iron supple-
mentation does not affect methanogenic activity. Indeed, the minimum
free iron requirement for methanogenic bacteria is available during all
the simulation tests. On the other hand, the increase of iron supply
directly results in the increase of iron sulphide concentration, with
consequent precipitation of sulfur compounds and a significant de-
crease of H2S gas production during AD of FW.
The potential use of the presented model as a predictive tool for TE
deficiency in AD has been further highlighted by investigating the effect
of lower external iron additions (in the range of
0.000005–0.0000001 mol L−1) to the simulated bioreactor (Fig. 4).
Due to extremely low free iron concentrations, methane production
profiles collapsed after 85, 100 and 130 days when the initial iron
concentration was set to 0.000001, 0.0000005 and 0.0000001 mol L−1,
respectively. According to this evidence, iron sulphide concentration
and precipitation was not significant and very low H2S gas production
were observed before the AD reactor failure. Noteworthy, to definitely
reach an appropriate calibration and validation of the model for the
prediction of real scale reactor failures under very low TE concentration
regime, sophisticated experimental techniques for the data collection
and ad-hoc experimental activities are still required.
5. Conclusion
A mathematical model able to consider the complex dynamics of
TEs in anaerobic systems has been presented. The model was im-
plemented as an original extension of ADM1 and it is able to account for
the main processes affecting TE evolution in AD bio-reactors. Two
different applications have been presented: the first relates to a real
experimental case where TE starvation occurs during the AD of FW. The
second application has been aimed at evaluating the biogas composi-
tion and hydrogen sulphide production due to different TE external
additions. Due to the complex interactions among the biological, che-
mical and physicochemical processes involved in TE speciation, the
model can be used as a predictive tool to obtain a deeper understanding
of TE bioavailability and to develop an optimal supplementation
strategy for full scale applications.

600
L. Frunzo, et al. Journal of Environmental Management 241 (2019) 587–602

CH 4 production Free Iron concentration

1.5 10-3

Fe2+ [Mol L -1]

CH 4 [LL -1d -1]
1 10-4

0.5 10-5

0 10-6
-100 0 100 200 -100 0 100 200
Time [d] Time [d]

H 2S production
10-3 10-7 Iron Sulfide concentration
2 1.5

1.5
H 2S [LL -1 d-1]

FeS [Mol L -1]

1

0.5
0.5

0 0
-100 0 100 200 -100 0 100 200
Time [d] Time [d]
0.000005M Fe In 0.000001M Fe In 0.0000005M Fe In 0.0000001M Fe In

Fig. 4. Methane production, free iron concentration, hydrogen sulphide gas production and iron sulphide concentration with lowering iron supplementation.

Acknowledgments
This work has been developed in the framework of the COST Action
1302 (“European Network on Ecological Roles of Trace Metals in
Anaerobic Biotechnologies”) supported by COST (European
Cooperation in Science and Technology). L. Frunzo and M.R. Mattei
acknowledge the support from the Progetto Giovani G.N.F.M. 2017
"Analisi di Sistemi Biologici Complessi". L. Frunzo also acknowledges
the support from the project VOLAC - Valorization of OLive oil wastes
for sustainable production of biocide-free Antibiofilm Compounds,
funded by CARIPLO foundation.
References
Bachmann, S., Wentzel, S., Eichler-Löbermann, B., 2011. Codigested dairy slurry as a
phosphorus and nitrogen source for Zea mays L. and Amaranthus cruentus L. J. Plant
Nutr. Soil Sci. 174 (6), 908–915.
Barrera, E.L., Spanjers, H., Solon, K., Amerlinck, Y., Nopens, I., Dewulf, J., 2015.
Modeling the anaerobic digestion of cane-molasses vinasse: extension of the
Anaerobic Digestion Model No. 1 (ADM1) with sulfate reduction for a very high
strength and sulfate rich wastewater. Water Res. 71, 42–54.
Batstone, D.J., 2006. Mathematical modelling of anaerobic reactors treating domestic
wastewater: rational criteria for model use. Rev. Environ. Sci. Bio/Technol. 5 (1),
57–71.
Batstone, D.J., Keller, J., 2003. Industrial applications of the IWA anaerobic digestion
model No. 1 (ADM1). Water Sci. Technol. 47 (12), 199–206.
Batstone, D.J., et al., 2002. The IWA anaerobic digestion model no 1 (ADM1). Water Sci.
Technol. 45, 65–73.
Blumensaat, F., Keller, J., 2005. Modelling of two-stage anaerobic digestion using the
IWA Anaerobic Digestion Model No. 1 (ADM1). Water Res. 39 (1), 171–183.
Callander, I.J., Barford, J.P., 1983a. Precipitation, chelation, and the availability of me-
tals as nutrients in anaerobic digestion. I. Methodology. Biotechnol. Bioeng. 25 (8),
1947–1957.
Callander, I.J., Barford, J.P., 1983b. Precipitation, chelation, and the availability of me-
tals as nutrients in anaerobic digestion. II. Applications. Biotechnol. Bioeng. 25 (8),
1959–1972.
Cao, W., Wang, M., Liu, M., Zhang, Z., Sun, Z., Miao, Y., Hu, C., 2018. The chemical and
dynamic distribution characteristics of iron, cobalt and nickel in three different
anaerobic digestates: effect of pH and trace elements dosage. Bioresour. Technol.
269, 363–374.
Cheng, X., Wang, J., Chen, B., Wang, Y., Liu, J., Liu, L., 2017. Effectiveness of phosphate
removal during anaerobic digestion of waste activated sludge by dosing iron (III). J.
Environ. Manag. 193, 32–39.
Choong, Y.Y., Norli, I., Abdullah, A.Z., Yhaya, M.F., 2016. Impacts of trace element
supplementation on the performance of anaerobic digestion process: a critical review.
Bioresour. Technol. 209, 369–379.
Comte, S., Guibaud, G., Baudu, M., 2006. Biosorption properties of extracellular poly-
meric substances (EPS) resulting from activated sludge according to their type: so-
luble or bound. Process Biochem. 41 (4), 815–823.
D'Acunto, B., Esposito, G., Frunzo, L., Mattei, M.R., Pirozzi, F., 2015. Mathematical
modeling of heavy metal biosorption in multispecies biofilms. J. Environ. Eng. 142
(9), C4015020.
D'Acunto, B., Frunzo, L., Mattei, M.R., 2018. On a free boundary problem for biosorption
in biofilms. Nonlinear Anal. Real World Appl. 39, 120–141.
De Gracia, M., Sancho, L., García-Heras, J.L., Vanrolleghem, P., Ayesa, E., 2006. Mass and
charge conservation check in dynamic models: application to the new ADM1 model.
Water Sci. Technol. 53 (1), 225–240.
De-Bashan, L.E., Bashan, Y., 2004. Recent advances in removing phosphorus from was-
tewater and its future use as fertilizer (1997–2003). Water Res. 38 (19), 4222–4246.
Esposito, G., Frunzo, L., Panico, A., Pirozzi, F., 2011a. Modelling the effect of the OLR and
OFMSW particle size on the performances of an anaerobic co-digestion reactor.
Process Biochem. 46 (2), 557–565.
Esposito, G., Frunzo, L., Panico, A., Pirozzi, F., 2011b. Model calibration and validation
for OFMSW and sewage sludge co-digestion reactors. Waste Manag. 31 (12),
2527–2535.
Esposito, G., Frunzo, L., Giordano, A., Liotta, F., Panico, A., Pirozzi, F., 2012. Anaerobic
co-digestion of organic wastes. Rev. Environ. Sci. Bio/Technol. 11 (4), 325–341.
Fedorovich, V., Lens, P., Kalyuzhnyi, S., 2003. Extension of enaerobic digestion model No.

601
L. Frunzo, et al.
1 with processes of sulfate reduction. Appl. Biochem. Biotechnol. 109 (1–3), 33–45.
Fermoso, F.G., Bartacek, J., Jansen, S., Lens, P.N.L., 2009. Metal supplementation to
UASB bioreactors: from cell-metal interactions to full-scale application. Sci. Total.
Environ. 407 (12), 3652–3667.
Fermoso, F.G., Collins, G., Bartacek, J., O'Flaherty, V., Lens, P., 2008. Acidification of
methanol-fed anaerobic granular sludge bioreactors by cobalt deprivation: induction
and microbial community dynamics. Biotechnol. Bioeng. 99 (1), 49–58.
Fermoso, F.G., Van Hullebusch, E.D., Guibaud, G., Collins, G., Svensson, B.H., Carliell-
Marquet, C., Frunzo, L., 2015. Fate of trace metals in anaerobic digestion. In: Biogas
Science and Technology. Springer International Publishing, pp. 171–195.
Flemming, H.C., Wingender, J., 2010. The biofilm matrix. Nat. Rev. Microbiol. 8 (9), 623.
Flores-Alsina, X., Solon, K., Mbamba, C.K., Tait, S., Gernaey, K.V., Jeppsson, U., Batstone,
D.J., 2016. Modelling phosphorus (P), sulfur (S) and iron (Fe) interactions for dy-
namic simulations of anaerobic digestion processes. Water Res. 95, 370–382.
Frunzo, L., 2017. Modeling Sorption of Emerging Contaminants in Biofilms. arXiv pre-
print arXiv:1706.04541.
Frunzo, L., Esposito, G., Pirozzi, F., Lens, P., 2012. Dynamic mathematical modeling of
sulfate reducing gas-lift reactors. Process Biochem. 47 (12), 2172–2181.
Gadd, G.M., 2009. Biosorption: critical review of scientific rationale, environmental im-
portance and significance for pollution treatment. J. Chem. Technol. Biotechnol. 84
(1), 13–28.
Gerardi, M.H., 2006. Wastewater Bacteria, vol. 5 John Wiley & Sons.
Glass, J., Orphan, V.J., 2012. Trace metal requirements for microbial enzymes involved in
the production and consumption of methane and nitrous oxide. Front. Microbiol.
3, 61.
Gonzalez-Gil, G., Kleerebezem, R., Lettinga, G., 1999. Effects of nickel and cobalt on
kinetics of methanol conversion by methanogenic sludge as assessed by on-line CH4
monitoring. Appl. Environ. Microbiol. 65 (4), 1789–1793.
Gustavsson, J., 2012. Cobalt and Nickel Bioavailability for Biogas Formation. PhD
Thesis. University of Linkoping, Sweden.
Gustavsson, J., Yekta, S.S., Sundberg, C., Karlsson, A., Ejlertsson, J., Skyllberg, U.,
Svensson, B.H., 2013. Bioavailability of cobalt and nickel during anaerobic digestion
of sulfur-rich stillage for biogas formation. Appl. Energy 112, 473–477.
Hilton, B.L., Oleszkiewicz, J.A., 1988. Sulfide-induced inhibition of anaerobic digestion.
J. Environ. Eng. 114 (6), 1377–1391.
Hu, Q.H., Li, X.F., Du, G.C., Chen, J., 2008. Effect of nitrilotriacetic acid on bioavailability
of nickel during methane fermentation. Chem. Eng. J. 143 (1–3), 111–116.
Jansen, S., Gonzalez-Gil, G., van Leeuwen, H.P., 2007. The impact of Co and Ni speciation
on methanogenesis in sulfidic media-biouptake versus metal dissolution. Enzym.
Microb. Technol. 40 (4), 823–830.
Keating, C., Chin, J.P., Hughes, D., Manesiotis, P., Cysneiros, D., Mahony, T., Smith, C.J.,
Mc Grath, J.W., O’Flaherty, V., 2016. Biological phosphorus removal during high-
rate, low-temperature, anaerobic digestion of wastewater. Front. Microbiol. 7, 226.
Kodera, H., Hatamoto, M., Abe, K., Kindaichi, T., Ozaki, N., Ohashi, A., 2013. Phosphate
recovery as concentrated solution from treated wastewater by a PAO-enriched bio-
film reactor. Water Res. 47 (6), 2025–2032.
Langerak, E.P.A.V., Beekmans, M.M.H., Beun, J.J., Hamelers, H.V.M., Lettinga, G., 1999.
Influence of phosphate and iron on the extent of calcium carbonate precipitation
during anaerobic digestion. J. Chem. Technol. Biotechnol. Int. Res. Process Environ.
Clean Technol. 74 (11), 1030–1036.
Lee, J., Park, K.Y., Cho, J., Kim, J.Y., 2018. Releasing characteristics and fate of heavy
metals from phytoremediation crop residues during anaerobic digestion.
Chemosphere 191, 520–526.
Lin, C.Y., 1993. Effect of heavy metals on acidogenesis in anaerobic digestion. Water Res.
27 (1), 147–152.
Liu, Y., Wang, Q., Zhang, Y., Ni, B.J., 2015. Zero valent iron significantly enhances me-
thane production from waste activated sludge by improving biochemical methane
potential rather than hydrolysis rate. Sci. Rep. 5, 1–6. https://doi.org/10.1038/
srep08263.
Moestedt, J., Nordell, E., Yekta, S.S., Lundgren, J., Martí, M., Sundberg, C., Björn, A.,
2016. Effects of trace element addition on process stability during anaerobic co-di-
gestion of OFMSW and slaughterhouse waste. Waste Manag. 47, 11–20.
Möller, K., Müller, T., 2012. Effects of anaerobic digestion on digestate nutrient avail-
ability and crop growth: a review. Eng. Life Sci. 12 (3), 242–257.
Möller, K., Stinner, W., 2010. Effects of organic wastes digestion for biogas production on
mineral nutrient availability of biogas effluents. Nutrient Cycl. Agroecosyst. 87 (3),
395–413.
Mudhoo, A., Kumar, S., 2013. Effects of heavy metals as stress factors on anaerobic di-
gestion processes and biogas production from biomass. Int. J. Environ. Sci. Technol.
10 (6), 1383–1398.
Ortner, M., Rachbauer, L., Somitsch, W., Fuchs, W., 2014. Can bioavailability of trace
nutrients be measured in anaerobic digestion? Appl. Energy 126, 190–198.
602
Journal of Environmental Management 241 (2019) 587–602
Ortner, M., Rameder, M., Rachbauer, L., Bochmann, G., Fuchs, W., 2015. Bioavailability
of essential trace elements and their impact on anaerobic digestion of slaughterhouse
waste. Biochem. Eng. J. 99, 107–113.
Papirio, S., Frunzo, L., Mattei, M.R., Ferraro, A., Race, M., D'Acunto, B., Esposito, G.,
2017. Heavy metal removal from wastewaters by biosorption: mechanisms and
modeling. In: Sustainable Heavy Metal Remediation. Springer, Cham, pp. 25–63.
Romero-Güiza, M.S., Vila, J., Mata-Alvarez, J., Chimenos, J.M., Astals, S., 2016. The role
of additives on anaerobic digestion: a review. Renew. Sustain. Energy Rev. 58,
1486–1499.
Rosén, C., Jeppsson, U., 2006. Aspects on ADM1 Implementation within the BSM2
Framework. Department of Industrial Electrical Engineering and Automation, Lund
University, Lund,, Sweden, pp. 1–35.
Roussel, J., Carliell-Marquet, C., 2016. Significance of vivianite precipitation on the
mobility of iron in anaerobically digested sludge. Front. Environ. Sci. 4, 1–12.
https://doi.org/10.3389/fenvs.2016.00060.
Saidou, H., Korchef, A., Moussa, S.B., Amor, M.B., 2009. Struvite precipitation by the
dissolved CO2 degasification technique: impact of the airflow rate and pH.
Chemosphere 74 (2), 338–343.
Sanchez, E., Borja, R., Weiland, P., Travieso, L., Martin, A., 2000. Effect of temperature
and pH on the kinetics of methane production, organic nitrogen and phosphorus
removal in the batch anaerobic digestion process of cattle manure. Bioprocess Eng. 22
(3), 247–252.
Shi, W., Chance, M.R., 2011. Metalloproteomics: forward and reverse approaches in
metalloprotein structural and functional characterization. Curr. Opin. Chem. Biol. 15
(1), 144–148.
Thanh, P.M., Ketheesan, B., Yan, Z., Stuckey, D., 2016. Trace metal speciation and
bioavailability in anaerobic digestion: a review. Biotechnol. Adv. 34 (2), 122–136.
Thanh, P.M., Ketheesan, B., Yan, Z., Stuckey, D., 2017. Effect of Ethylenediamine-N, N′-
disuccinic acid (EDDS) on the speciation and bioavailability of Fe2+ in the presence
of sulfide in anaerobic digestion. Bioresour. Technol. 229, 169–179.
van Hullebusch, E.D., Guibaud, G., Simon, S., Lenz, M., Yekta, S.S., Fermoso, F.G., Jain,
R., Duester, L., Roussel, J., Guillon, E., Skyllberg, U., Almeida, C.M.R., Pechaud, Y.,
Garuti, M., Frunzo, L., Esposito, G., Carliell-Marquet, C., Ortner, M., Collins, G., 2016.
Methodological approaches for fractionation and speciation to estimate trace element
bioavailability in engineered anaerobic digestion ecosystems: an overview. Critic.
Rev. Eviron. Sci. Technol. 46 (16), 1324–1366.
Van Hullebusch, E.D., Zandvoort, M.H., Lens, P.N., 2003. Metal immobilisation by bio-
films: mechanisms and analytical tools. Rev. Environ. Sci. Biotechnol. 2 (1), 9–33.
Vintiloiu, A., Boxriker, M., Lemmer, A., Oechsner, H., Jungbluth, T., Mathies, E.,
Ramhold, D., 2013. Effect of ethylenediaminetetraacetic acid (EDTA) on the bioa-
vailability of trace elements during anaerobic digestion. Chem. Eng. J. 223, 436–441.
Wang, J., Chen, C., 2009. Biosorbents for heavy metals removal and their future.
Biotechnol. Adv. 27 (2), 195–226.
Wintsche, B., Glaser, K., Sträuber, H., Centler, F., Liebetrau, J., Harms, H., Kleinsteuber,
S., 2016. Trace elements induce predominance among methanogenic activity in
anaerobic digestion. Front. Microbiol. 7, 2034.
Ye, M., Liu, J., Ma, C., Li, Y.Y., Zou, L., Qian, G., Xu, Z.P., 2018. Improving the stability
and efficiency of anaerobic digestion of food waste using additives: a critical review.
J. Clean. Prod. 192, 316–326.
Yekta, S.S., Svensson, B.H., Björn, A., Skyllberg, U., 2014. Thermodynamic modeling of
iron and trace metal solubility and speciation under sulfidic and ferruginous condi-
tions in full scale continuous stirred tank biogas reactors. Appl. Geochem. 47, 61–73.
Yekta, S.S., Skyllberg, U., Danielsson, Å., Björn, A., Svensson, B.H., 2017. Chemical
speciation of sulfur and metals in biogas reactors–Implications for cobalt and nickel
bio-uptake processes. J. Hazard. Mater. 324, 110–116.
Zandvoort, M.H., Van Hullebusch, E.D., Fermoso, F.G., Lens, P.N.L., 2006. Trace metals in
anaerobic granular sludge reactors: bioavailability and dosing strategies. Eng. Life
Sci. 6 (3), 293–301.
Zhang, L., Jahng, D., 2012. Long-term anaerobic digestion of food waste stabilized by
trace elements. Waste Manag. 32 (8), 1509–1515.
Zhang, L., Keller, J., Yuan, Z., 2009. Inhibition of sulfate-reducing and methanogenic
activities of anaerobic sewer biofilms by ferric iron dosing. Water Res. 43 (17),
4123–4132.
Zhang, Y., Piccard, S., Zhou, W., 2015a. Improved ADM1 model for anaerobic digestion
process considering physico-chemical reactions. Bioresour. Technol. 196, 279–289.
Zhang, W., Wu, S., Guo, J., Zhou, J., Dong, R., 2015b. Performance and kinetic evaluation
of semi-continuously fed anaerobic digesters treating food waste: role of trace ele-
ments. Bioresour. Technol. 178, 297–305.
Zhang, W., Zhang, L., Li, A., 2015c. Enhanced anaerobic digestion of food waste by trace
metal elements supplementation and reduced metals dosage by green chelating agent
[S, S]-EDDS via improving metals bioavailability. Water Res. 84, 266–277.