Bioresource Technology 341 (2021) 125795

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

Review

Agro-industrial wastewater in a circular economy: Characteristics, impacts

and applications for bioenergy and biochemicals
Walter José Martinez-Burgos a, Eduardo Bittencourt Sydney b,
Adriane Bianchi Pedroni Medeiros a, Antonio Irineudo Magalhães a, Júlio Cesar de Carvalho a,
Susan Grace Karp a, Luciana Porto de Souza Vandenberghe a, b, Luiz Alberto Junior Letti a,
Vanete Thomaz Soccol a, Gilberto Vinícius de Melo Pereira a, Cristine Rodrigues a,
Adenise Lorenci Woiciechowski a, Carlos Ricardo Soccol a, *
a
Federal University of Paraná, Department of Bioprocess Engineering and Biotechnology, Centro Politécnico, 81531-990, Curitiba Paraná, Brazil
b
Federal University of Technology – Paraná, Department of Bioprocess Engineering and Biotechnology, 84016-210, Ponta Grossa Paraná, Brazil

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

• Agro-industrial effluents have signifi­

cant environmental impacts.
• Large volumes of agro-industrial efflu­
ents are generated worldwide by agro-
industries.
• Bioenergy is the most promising way to
valorize agro-industrial wastewater.
• Obtaining biometabolites from effluents
is an alternative to increase profitability.

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

Keywords: The generation of agroindustrial byproducts is rising fast worldwide. The slaughter of animals, the production of
Agribusiness bioethanol, and the processing of oil palm, cassava, and milk are industrial activities that, in 2019, generated
Effluents huge amounts of wastewaters, around 2448, 1650, 256, 85, and 0.143 billion liters, respectively. Thus, it is
Environmental impacts
urgent to reduce the environmental impact of these effluents through new integrated processes applying bio­
Bioenergy
Biochemicals
refinery and circular economy concepts to produce energy or new products. This review provides the charac­
teristics of some of the most important agro-industrial wastes, including their physicochemical composition,
worldwide average production, and possible environmental impacts. In addition, some alternatives for reusing
these materials are addressed, focusing mainly on energy savings and the possibilities of generating value-added
products. Finally, this review considers recent research and technological innovations and perspectives for the
future.

1. Introduction agricultural and forestry products, either for food or non-food purposes
(Rosete, 2020). The agroindustry is the main responsible for the pro­
Agribusiness can be defined as a set of economic activities including duction and processing of food and feed, but also the production of raw
production, processing or industrialization, and commercialization of material for other industrial areas. The agriculture and industrial sectors

* Corresponding author.
E-mail address: soccol@ufpr.br (C.R. Soccol).

https://doi.org/10.1016/j.biortech.2021.125795
Received 2 July 2021; Received in revised form 11 August 2021; Accepted 13 August 2021
Available online 18 August 2021
0960-8524/© 2021 Published by Elsevier Ltd.
W.J. Martinez-Burgos et al. Bioresource Technology 341 (2021) 125795

are responsible for approximately 4% and 28%, respectively, of the Table 1

world’s gross domestic product (World-Bank, 2020). The main products Estimated amounts of agro-industrial effluents generated in the world.
of the sector are derived from sugarcane (alcohol and sugar), cassava Raw material Effluent Effluent generation Wastewater
(flour and starch), cereals (flour and alcohol), oilseeds (palm oil, soy processed ratio generated,
oil), milk and derivatives, and animal slaughter (cattle, pigs, and billion liters
poultry) (FAOSTAT, 2020; IndexMundi, 2020; Statista, 2020; USDA, 142 million tons in Cassava 600L CPW/ton 85
2020). These activities require large amounts of soil, energy and water, 20181( processing cassava (Carvalho
generating significant volumes of solid materials and wastewaters with FAOSTAT, 2020) wastewater (CPW) et al., 2018;
Martinez-Burgos
important polluting characteristics (Saval, 2012). The generated efflu­ et al., 2019)
ents have high levels of organic matter, sometimes with toxic com­ 110 million m3 in Vinasse 15 L Vinasse/L 1650
pounds and even heavy metals, that can impact in the quality of water, 20192(Statista, wastewater (VW) ethanol (Madaleno
soil and air (Azman et al., 2016; Fuess and Garcia, 2014; Rivadeneyra- 2020) et al., 2020; Sydney
et al., 2021)
Domínguez et al., 2013). The high eutrophic and polluting potential of
73 million tons oil Palm oil mill 3.75 ton POME/ton 256
agro-industrial wastewaters requires chemical and biological treatment in 2019 ( effluent (POME) crude palm oil (
before being discarded in the environment (Ferreira et al., 2021). IndexMundi, Cheng et al., 2018)
The physical, chemical, and biological treatment of agro-industrial 2020)
wastes is not an ideal solution. These effluents have the potential to be 720 million tons in Effluent of milk 0.2 L effluent/L 143
2018(FAOSTAT, processing (EMP) processed milk (
used as feedstock and processed to generate bioenergy and other 2020) Vourch et al.,
different bioproducts with significant economic value. This strategy of 2008)
maximizing the use of natural resources by significantly reducing 21.5 million tons in Cheese whey (CW) 0.9 m3 whey/m3 158
environmental impacts is a current demand of society (Dahiya et al., 2014(FAOSTAT, milk processed (
2020) Siso, 1996)
2018). Agro-industrial wastes are generated at high rates and volumes
98.4 million tons of Slaughterhouse 10 m3/ton chicken 984
and have been extensively studied to produce metabolites of industrial chicken effluents (SH) carcasses (Fagnani
interest, such as organic acids, alcohols, proteins, microbial biomass, carcasses in 2019 et al., 2019; USDA,
methane, biohydrogen, biobutanol, etc. (Dahiya et al., 2018; Rosa et al., (USDA, 2020) 2020)
2020). The application of biobased economy has increased greatly in 108.5 million tons 8.3 m3/ton cattle 900
of pig carcasses carcass (Aziz et al.,
recent years, and the identification of the most promising pathways is
in 2019 (USDA, 2019;USDA, 2020)
important to support scientific and technological development for waste 2020)
valorization. This review article focuses on agro-industrial wastewater 62.6 million tons of 9 m3/ton cattle 564
generated in animal slaughter, in the production of sugarcane ethanol, cattle carcasses carcass (Aziz et al.,
in 2019 (USDA, 2019; USDA, 2020)
palm oil, cassava starch, and in milk processing. A critical analysis of the
2020)
most promising strategies was carried out, focusing on risk reduction
and the development of a biobased economy, for the bioconversion of
agro-industrial wastewaters. So, this review presents the panorama of process. In the case of CW, POME, CPW, and VW, the effluents are direct
the amount of generated agro-industrial effluents in the world, their outflows of the process; other effluents such SH and EMP are generated
environmental impacts and the amount of energy that could be gener­ practically during the whole process as a result of cleaning and sani­
ated and/or recovered. tizing operations, as shown in Fig. 1. In the case of SH, it is also
generated by washing the carcasses of the animals; SH can also contain
2. Agro-industrial effluents generated worldwide animal vomit and urine (Fig. 1A). The CW is the liquid fraction obtained
during the coagulation of milk in the cheese manufacture process, after
The slaughter of animals, the production of fuel alcohols and vege­ the separation of the coagulum or micellar phase (Xia et al., 2020), and
table oils, and the processing of milk and cassava are industrial activities can be considered an effluent or not. In the case of POME, an effluent
that generate large amounts of effluents worldwide, as shown in Table 1. produced by the processing of oil palm, it is generated in three process
In 2019, 2.45 and 1.65 billion m3 of effluents were generated at the steps: sterilization, separation, and hydrocyclones (Fig. 1C). On average,
slaughterhouses and the biofuel plants, respectively. According to Aziz the amount of POME generated in these steps is 0.9–1.5, 1.5–2.0 and
et al.(2019), Fagnani et al. (2019) and Mburu et al. (2019) the slaughter 0.1–0.5 tons / ton of crude palm oil, respectively (Ahmed et al., 2015).
of animals is especially worrying for the large amounts of effluents CPW and VW are generated in the centrifugation and distillation steps,
generated and their pollution potential. The main countries that respectively, as represented in Fig. 1D and 1E (Christofoletti et al., 2013;
generate wastes from animals, here named slaughterhouse effluents Moraes et al., 2015; Tran et al., 2015), CPW is the water separated from
(SH), are China, US, European Union, and Brazil, with 24%, 17%, 16%, the starch, either by centrifugation or decanting processes, and VW is the
and 10% of contribution, respectively. In the same way, US, Brazil, the liquid generated when separating ethanol from the fermented wort.
European Union, and China are the main generators of vinasse waste­ The physicochemical characteristics of the effluents are quite
water (VW) from bioethanol production, with 54%, 30%, 5%, and 3% of diverse, even among those of the same origin. For example, in the case of
contribution, respectively. The main producers of cassava processing POME, the chemical oxygen demand (COD) ranged from approximately
wastewater (CPW) are Nigeria, Thailand, the Democratic Republic of 21 to 75 gO2/L, the biochemical oxygen demand (BOD) varied between
Congo and Ghana with 21%, 11%, 10% and 7% of participation, 23 and 42 gO2/L, and the phosphorus content from 7 to 170 mg/L.
respectively. Large volumes of palm oil mill effluent (POME), around Similarly, high variability in other parameters was observed in all ef­
250 billion liters, are also generated worldwide, approximately 82% fluents, as illustrated in Fig. 2. The variability can be attributed to the
originated from Indonesia and Malaysia. Cheese whey (CW) and the type of process and technology and to the characteristics of the raw
effluent of milk processing (EMP) are generated in highest volumes in material: for plants, the variety, nutrition, season, etc., and for animals,
the United States, India, and Brazil. the breed, feed, and environmental conditions (Aziz et al., 2019). The
effluents CPW, VW, CW, and POME contain the highest organic loads
3. Agro-industrial processes, generation of effluents, and their (COD), an average of 55 g O2/L (Fig. 2A). This is explained by the fact
characteristics that these effluents contain high levels of carbohydrates, proteins, and
fat.
Effluents can be generated in different steps or in a single step of the

2
W.J. Martinez-Burgos et al.
Although SH and EMP have the lowest organic loads, around 5 g O2/
L, this value is still considered extremely high, because the environ­
mental legislation has established that COD should not exceed 50 mg
O2/L, associated to pH values between 5 and 9 and a total nitrogen
content below 200 ppm (Hameed et al., 2003). For example, it was
observed that the effluents with a pH closer to neutrality were SH and
EMP, and the most acidic were VW and CW with a pH around 4.5
(Fig. 2E). In this sense, all these effluents need to be treated before being
disposed into the environment. Generally, the temperatures of these
3
Bioresource Technology 341 (2021) 125795
Fig. 1. Simplified process of: A) Slaughterhouse
for animals and slaughterhouse effluents (SH)
generated, adapted from Scarassati et al. (2003); B)
Milk processing, effluent of milk processing (EMP)
generation, and cheese whey (CW), adapted from
FEAM (2013); C) Extraction of palm oil and palm
oil mill effluent (POME) generation, adapted from
Ahmed et al. (2015); D) Starch or cassava flour
processing and generation of cassava processing
waste (CPW), adapted from Tran et al. (2015); E)
Alcohol production and vinasse wastewater (VW)
generation, adapted from Christofoletti et al. (2013)
and Rodrigues Reis and Hu (2017). The operations in
the boxes with oblique lines require energy. Dis­
continued lines indicate effluents that can be used to
generate bioenergy, or other types of metabolites.
*The energy content of each effluent was estimated
using equation 1.
effluents are between 20 and 35 ◦ C, except POME and VW, that leave the
process with temperatures around 85 ◦ C (Moraes et al., 2015; Norfadilah
et al., 2016), and must be cooled before being disposed of.
The energy content of the effluents is mainly attributed to the con­
tent of organic matter, which is used by microorganisms as substrates for
the production of methane or hydrogen; in this work, the calculations
were made considering methane as an energy carrier. Thus, POME,
CPW, CW, and VW are the effluents with the highest energy contents of
281, 195, 155, and 132 kWh/m3, respectively. Although SH and EMP
W.J. Martinez-Burgos et al.
are the wastewaters with the lowest energy content, their use could be
justified by the high volumes generated.
In terms of available substrates, CW contains significant concentra­
tions of lactose, galactose and glucose (36.5 g L-1, 2.31 g L-1 and 1.12 g L-
1
, respectively, as reported by Nielsen et al. (2021) and low fat con­
centrations in the range of 0.1–1.14% (Martínez-Ruano et al., 2019).
EMP contains low concentration of lipids and significant concentration
of lactose. CPW is also a source of carbohydrates (total carbohydrates:
58.2–5.2 g L− 1), reducing sugars (38–0.57 g L− 1), and non-reducing
sugars (22.2–14.98 g L− 1) (Carvalho et al., 2018). The main carbon
sources present in this effluent are starch and maltose (Martinez-Burgos
et al., 2020). POME and SH contain low concentrations of fermentable
carbohydrates (0.23 g L-1 and 0.081 g L-1, respectively) but high levels of
fat (37 g L− 1 and 8.9 g L-1, respectively). However, POME contains
hemicellulose (2.5–8.4 g L-1) and cellulose (3.9–4.3 g L-1), which can be
hydrolyzed to fermentable carbohydrates (Cheng et al., 2021). VW
contains low concentrations of glucose (0.01–0.64 gL-1) and fructose
(0.02–0.77 gL-1), but it does contain reasonable amounts of organic
acids such as lactic acid (0.61–6.45 g L− 1), succinic acid (0.09–3.44 g
L− 1), acetic acid (0.06–3.80 g L− 1), and aconitic acid (0.21–3.79 g L− 1)
(Godoi et al., 2019). CW also contains appreciable amounts of lactic acid
(Martínez-Ruano et al., 2019). These compounds also affect COD and
BOD5, and organic acids are also used as alternative carbon sources by
microorganisms (do Nascimento Junior et al., 2021; Martinez-Burgos
et al., 2021a).
The effluents also contain significant concentrations of other nutri­
ents, such as nitrogen (Fig. 2C) and phosphorus (Fig. 2D), and micro­
nutrients, such as Ca, S, K and Mg (Aziz et al., 2019; Barros et al., 2008;
Nitayavardhana et al., 2013; Norfadilah et al., 2016). Some effluents
also contain proteins. For example, protein concentrations were re­
ported for SH (4.8 g L-1) (Haselroth et al., 2021), CPW (1.5 g L-1) (Car­
valho et al., 2018), POME (460–1580 mg L-1) (Cheng et al., 2021) and
CW (1% w/w) (Damar et al., 2020). Furthermore, the effluents are
sources of amino acids such as alanine, histidine, glycine, isoleucine,
4
Bioresource Technology 341 (2021) 125795
Fig. 2. Variability of the physicochemical
composition of effluents. A) COD; B) BOD; C) N; D)
P; E) pH. CPW: cassava processing waste; VW:
vinasse wastewater; SH: slaughterhouse effluent;
EMP: effluent of milk processing; CW: cheese whey;
POME: palm oil mill effluent; COD: chemical oxygen
demand; BOD: biochemical oxygen demand. Sources:
(Damasceno et al., 2003; Nitschke and Pastore, 2003;
Nitschke and Pastore, 2006; Maróstica and Pastore,
2006; Aguilar et al., 2002; Ahmed et al., 2015; Barros
et al., 2008; Bhatia et al., 2007; Bustillo-Lecompte
and Mehrvar, 2015, 2016; Gelegenis et al., 2007;
Nitayavardhana et al., 2013; Norfadilah et al., 2016;
Robles-González et al., 2012; Rupani et al., 2010;
Saddoud et al., 2007; Singh et al., 2013; Sir­
ianuntapiboon et al., 2005; Sun et al., 2012; Tawfik
et al., 2008; Venetsaneas et al., 2009; Rajab et al.,
2017; Ohimain et al., 2013; Ohimain et al., 2012;
Wood et al., 1979; Awotoye et al., 2011; Borja et al.,
1996; Martinez-Burgos et al., 2021b; Xia et al., 2020;
Godoi et al., 2019; Cheng et al., 2021).
leucine, aspartic acid, glutamic acid, lysine, methionine, proline, serine,
threonine, tryptophan, tyrosine and valine (Cheng et al., 2021; Damar
et al., 2020; da Silva et al., 2017). Some vitamins such as vitamin B2 and
vitamin C have also been found in CPW (Li et al., 2018).
Finally, some of these effluents also contain heavy metals which can
limit their use. For example, VW is one of the effluents with the most
variety of heavy metal traces. According to (Godoi et al., 2019) this
effluent contains As2+,Cd2+,Cr3+, Co2+, Cu2+, Pb2+, Hg2+,Ni2+ and
Zn2+. Traces of Cd2+, Cr3+, Pb2+, Hg2+, Ni2+ and As2+ have been found
in POME (Cheng et al., 2021) and in CPW traces of Cd2+, Cr3+, Cu2+,
Pb2+ and Hg2+ were reported (Olaoye et al., 2020).
4. Environmental impacts of agro-industrial effluents
Generally, agro-industrial effluents contain significant amounts of
phosphorus and nitrogen; so, when they are discarded in water bodies,
they promote the growth of microorganisms, aquatic plants, and
microalgae. In most cases, the emissions resulting from these effluents
promote eutrophication processes, which affect some species and
destabilize ecosystems completely, making the receiving water bodies
improper for other uses (Silva et al., 2005). Effluents can also impact the
soil when they are used in the fertigation. An example of the indis­
criminate use of effluents in fertigation is CPW that increases the salt
content in the soil (Abegunrin et al., 2016). The VW provides large
amounts of chlorides and nitrates that can reach the groundwater
through leaching processes (Fuess and Garcia, 2014), which can cause
soil contamination by toxic ions and metals. Another risk associated
with the use of wastewaters in fertigation is the hydrophobicity that the
soil can acquire, which is generated by the organic matter and the mi­
croorganisms covering the surface (Vogelmann et al., 2013). The
organic overload can clog the pores of the soil, decrease the dissolved
oxygen concentration, and exchange the microbial population for an
undesirable one (Fuess and Garcia, 2014).
Another worrying impact of effluents is the generation of different
W.J. Martinez-Burgos et al. Bioresource Technology 341 (2021) 125795

greenhouse gases. An example is the organic matter of CPW that can be
biodegraded to CH4 and CO2, gases with potent greenhouse effect (Ayu
and Aryati, 2010; Silva et al., 2005), which can be released into the
atmosphere when effluents are improperly disposed. Effluents such as
POME, EMP, VW, and SH also generate greenhouse gases, and the latter
can also release NOx and SOx, responsible for acid rain (Danesh Miah
et al., 2010; Yang et al., 2012). Carmo et al. (2013) found that sugarcane
crops fertilized with VW had higher fluxes of both CO2 and NO2 with
values of approximately 38,000 and 40 mg m-2 day-1, respectively.
Table 2 summarizes other types of impacts associated with agro-
industrial effluents.
5. Integrated production of bioproducts from agro-industrial
effluents
Although they are highly costly, conventional treatments of effluents
still predominate (Martinez-Burgos et al., 2019). This is the case of
coagulation and flocculation processes using coagulants such as
Fe2(SO4)3 and Al2(SO4)3 for the removal of colloidal material, for
example, phosphorus in the form of phosphate, nitrogen in proteins,
peptide bonds, and amino groups (Aguilar et al., 2002; Bhatia et al.,
2007). The simple treatment of effluents is in discordance with new
perspectives of development of a circular economy where waste is
considered a potential raw material.
Anaerobic fermentation of these liquid wastes to produce biogas as
an energy source is often a viable alternative. Non-toxic effluents with
readily fermentable substrates, such as CW, VW, EMP and POME, can be
used as raw materials of many bioproducts (alcohols, organic acids,
solvents, pigments), however their use to produce food and pharma­
ceuticals ingredients must be carefully considered based on an exhaus­
tive study of composition and no examples of pilot/industrial production
is known. Known toxic effluents are CPW, which contains toxic com­
pounds that generate hydrocyanic acid (Carvalho et al., 2018), and SH,
for its high pathogenic microbial load, presence of residues of detergents
and heavy metals (Aziz et al., 2019).
The use of the effluents as biofertilizers is also promising, due to their
nutrient components. In fact, one of the first applications of effluents
was fertigation, i.e., the use of effluents as fertilizers in different types of
crops to supply soil deficiencies of nitrogen, phosphorus, potassium, etc.
CPW, VW, CW and POME have been used as fertilizers in banana, corn,
sugar cane and Schinus molle crops, with beneficial effects (Abou-dahab
et al., 2019; Carrilho et al., 2016; Gómez and Rodríguez, 2000). How­
ever, these practices have been strongly questioned by the effects that
effluents can have on the soil as well as on groundwater (Abegunrin
et al., 2016; Christofoletti et al., 2013; Magalhaes et al., 2015). CW may
contain high concentrations of salts and lactic acid, which affect soil
properties (Carvalho et al., 2013); POME can cause a nutrient imbalance
on the soil (Onyia et al., 2001); SH usually contains high loads of
pathogenic microorganisms (2.4 × 1010 MPN mL− 1 of total coliforms
and 7.70 × 109 MPN mL− 1 of thermotolerant coliforms) (Bustillo-
Lecompte and Mehrvar, 2015; Davarnejad and Nasiri, 2017; Massé and
Masse, 2000), and pig slaughterhouse effluents have hindered the
germination of cucumber and lettuce seeds (Gerber et al., 2017).
5.1. The potential of agro-industrial wastewaters for biogas production
Energy is a universal requirement for industrial processing and is
mostly the first-considered strategy for waste valorization (Waste-to-
Energy). Low-water content solid wastes are generally used for thermal
and electrical production through direct burning. The simplicity and low
cost of this technology challenge the modern technologies. On the other
side, liquid effluents can be converted to bioenergy by different trans­
formation processes. While aerobic bioprocesses result in the production
of microbial biomass and oxidated bioproducts (generally with low
energy content), anaerobic bioprocesses combine a partial reduction of
the effluent’s polluting potential with the production of more reduced
molecules that can be used for energy production. In this context,
methane and biohydrogen have received great attention. While methane
production is considered a mature technology that generates energy
(methane) and fertilizer (digestate) with such high yields and versatility
that make it a primary option for effluents rich in organic matter, bio­
hydrogen is a developing technology that enables the production of
compounds that can be used for energy (hydrogen) and in the chemical
industry (hydrogen and short-chain fatty acids) (Martinez-Burgos et al.,
2021b). Methane from biogas can be purified to biomethane and used in
vehicles replacing fossil-based natural gas, saving the consumption of
vehicular fuels in the agroindustry.
Biomethane production processes reached up to 98% removal of
COD (Faisal and Unno, 2001). According to Ahmed et al. (2015) and Ma
et al. (2019), the digestion of organic matter occurs in four steps: hy­
drolysis, acidogenesis, acetogenesis, and methanogenesis. Hydrogen
and methane are specifically produced in the acetogenesis and

Table 2
Potential environmental impacts generated by agro-industrial effluents.
Type of Impacts Effluents References
CPW VW SH POME CW

Presence of toxic compounds +cyanogenic glycosides Heavy metals –

+(19)Heavy –
(Carvalho et al., 2018)1(Olaoye et al., 2020)2 (Godoi
and (1, 14)Heavy metals (2) (7)
metals et al., 2019) 7 (Cheng et al., 2021)19
Changes in ecosystems +(5) +(7) +(12) +(19) +(24) (Kobya et al., 2006)12 (De Gioannis et al., 2014)24
Changes in soil physical and +(1) +(8) +(13) +(20) +(25) (Gerber et al., 2017)13 (Ali Amat et al., 2015)20 (
chemical properties Decesaro et al., 2021) 25
Changes in soil microbial +(3) +(8) +(13) –
+(26) (Fuess and Garcia, 2014)3 (Martinez-Burgos et al.,
population 2019)14 (Souza et al., 2013)8 (Marone et al., 2017)26
Eutrophication +(1) +(7) +(15) +(19) +(27) (Aguilar et al., 2002)15(Carvalho et al., 2013)27
Water resource depletion +(1) +(9) +(16) +(21) +(27) (Scarpare, 2013)9 (Gunkel et al., 2007)10 (Bustillo-
Increasing the temperature –
+(10) –
+(22) –
Lecompte and Mehrvar, 2015) 16 (Norfadilah et al.,
of the receiving water 2016)21 (Hossain et al., 2016)22
bodies
Unpleasant odors +(5) +(7) +17 +20 +(27) (Ayu and Aryati, 2010)5 (Blazy et al., 2015)17
Depletion of dissolved +(1) +(10) +(18) +(19) +(28)
oxygen in water
Disseminate endemic –
+(7) – – –
(Carmo et al., 2013)11 (Ware and Power, 2017)18 (
diseases Krishnan et al., 2016)23 (Kothari et al., 2017)28
Release of greenhouse gases CH4 (5) CO2,–N2O, CH4, SO2, +(23) +(28)CH4,
CH4 +(11) NH3 +(18) SOx, NOx

*(+) indicates that the effluent generates this impact and the number is the reference **(–) indicates that the effluent does not generate the impact or no related
research was found
CPW: cassava processing waste; VW: vinasse wastewater; SH: slaughterhouse effluent; POME: palm oil mill effluent; CW: cheese whey

5
W.J. Martinez-Burgos et al. Bioresource Technology 341 (2021) 125795

methanogenesis phases, respectively (Moraes et al., 2015). Anaerobic
digestion is an effective way of reusing the effluents, resulting also in the
production of organic acids, such as propionic acid (Haddadin et al.,
1996), lactic acid (Roukas and Kotzekidou, 1991), and acetate, besides
energy in the form of methane and hydrogen (Kothari et al., 2017).
However, the efficiency of digestion and biogas productivity depend, in
a large extent, on the structure of the microbial community, as well as on
environmental factors, such as pH and temperature, which can be
controlled depending on the type of bioreactor.
The species that have received the most attention in biomethane
production are those belonging to the genera Methanosaeta, Meth­
anosarcina, Methanobacterium, Methanobrevibacter, Methanosphaera,
Methanomicrobium, Methanogenium, Methanoplanus, Methanoculleus,
Methanospirillum, Methanomicrobiales, and Methanocorpusculum (Franke-
whittle et al., 2009; Smith and Ingram-smith, 2007). In the case of
biohydrogen, the most highlighted microbial genera are Bacillus, Clos­
tridium and Enterobacter (Pattra et al., 2008; Ramprakash and Muthu­
kumar, 2016). Hydrogen is generated as an intermediate metabolite
from anaerobic digestion that is used in the final stage of digestion by
methanogenic bacteria to produce methane. In other words, the pro­
duction of hydrogen or methane depends mainly on the microbial
population.
Different types of bioreactors have been designed for methane and
hydrogen production from effluents, such as up-flow anaerobic filtra­
tion, pond system, anaerobic filtration, anaerobic digester, fluidized bed
reactor, up flow anaerobic sludge blanket reactor (Poh and Chong,
2009), up-flow anaerobic sludge fixed-film (Najafpour et al., 2006),
expanded granular sludge bed, ultrasonicated membrane anaerobic
system, ultrasonic-assisted membrane anaerobic system, membrane
anaerobic system, modified anaerobic baffled bioreactor, among others
(Moraes et al., 2015; Ohimain and Izah, 2017). The main characteristics
of these bioreactors are presented in Table 3.
One of the most widely used systems for effluent treatment and
biogas production is the pond system; approximately 85% of POME
produced in Malaysia is treated with this technology (Poh and Chong,
2009). In starch production industries, CPW is mostly directed for biogas
production, which is used for heat production in adapted boilers. This
bioprocess is most frequently carried out in covered lagoon biodigesters,
with unoptimized conditions of feed rate, agitation and temperature
control (Watthier et al., 2019). Another globally employed system is the
anaerobic baffled reactor, commonly used for the treatment of domestic
sewage.
VW was applied in two up-flow anaerobic bioreactors to produce
methane, using sludge from swine wastewater as inoculum, and 0.18 L
CH4/g COD were obtained (Barros et al., 2016). When using a covalently
immobilized consortium of methanogenic strains, the organic load of
VW was reduced by 92%, obtaining a higher yield (0.33 m3 CH4/kg
COD) (Lalov et al., 2001). The first industrial unit of electricity pro­
duction from biogas using VW and filter cake as substrates was recently
implemented in Brazil (October 2020) by the Company Raízen (Guariba,
São Paulo). In 2020, 1,829,065,334 Nm3 of biogas were produced in the
Country, with 79% of the plants being operated with agro-industrial
waste (CIBIOGAS, 2021).
According to Feroldi et al. (2014), the effluent from animal slaugh­
terhouses has a high capacity to generate biogas from the organic load
and the autochthonous microbial flora. For example, 90% of COD was
removed, and 60 mL of methane were generated from 320 mL of cattle
and pig SH (Vidal et al., 2016). Besides, (Ware and Power, 2017) ob­
tained a yield of 594.59 mL CH4 g/VS (volatile solids) from poultry
slaughter effluent. On the other hand, high lipid and protein contents are
limiting factors for anaerobic digestion (inhibition of microbial growth
and ammonia loads, respectively) and represent technical obstacles.
Industrial biogas plants treating SH (co-digestion) are being operated
since 1997 in Sweden (Ek et al., 2011).
According to Ahmed et al. (2015), 1 m3 of POME can generate about
28 m3 of biogas. Other researchers have already signaled that POME

Table 3
Bioreactors used for wastewater treatment and biogas production (H2 and CH4).
Type of Bioreactor features Retention COD removal Advantages Drawbacks References
Bioreactor time (days) efficiency
(%)

Pond system Depending on the effluents the first 20–200 97 Low cost; CH4 concentration up Biogas production depends (Poh and
ponds are for cooling, followed by to 55%; simple and easy on weather stations; large Chong, 2009)
mixing, anaerobic and facultative. operation areas needed; difficult gas
The system can consist of up to 12 capture
ponds.
Anaerobic Bioreactor with different 15 94 CH4 concentration up to 63%; High cost (Borja and
filtration compartments (airtight vessel, pH and T control (◦ C) Banks, 1994)
digester, and temperature gauge)
Modified Device with different compartments 5–10 93.3 ± 3.8 CH4 concentration 67–71%; High cost (Sung et al.,
anaerobic (feed tank, magnetic stirrer, control of variables such as: T (◦ 2019)
baffled peristaltic pump, water jacket) C) and pH.
bioreactor
Fluidized bed Compact bed system, can be operated 70 98–80 Large contact area; high organic Needs large amounts of (Borja et al.,
reactor with up or down flow loading rates; high flow velocity input power 2001)
Up-flow System consisting mainly of pump, Up to 200 60–97 Equipment versatility, can be Needs long start-up period (Ohimain and
anaerobic sludge blanket, gas separator, gas used for high suspended solids (2–4 months) Izah, 2017)
sludge blanket collector, wastewater
(UASB)
Anaerobic Set of series of vertical baffles that 3–5 82–92 Simple and inexpensive Inconvenient, with high (Faisal and
baffled reactor force effluent to flow over them from equipment; no moving part or viscosity effluents Unno, 2001;
inlet to outlet mixing device; CH4 Wang et al.,
concentration 50–75% 2004)
Up-flow Hybrid bioreactor (UASB and up flow 4–40 89–97 Rapid biotransformation of Complex system that needs (Najafpour
anaerobic fixed film); it is usually composed of organic matter; CH4 large amounts of input et al., 2008)
sludge fixed- pumps, feed tanks, gas separators, concentration 62–82% power
film brackets, etc.
Continuous It is a mechanically agitated closed 4 60–83 Easy operation; CH4 Contains mechanical (Ohimain and
stirred tank tank reactor which increases the concentration 65–67% systems; needs significant Izah, 2017)
reactor (CSTR) contact area with biomass and amounts of energy
increases biogas production.

COD: chemical oxygen demand

6
W.J. Martinez-Burgos et al.
contains large amounts of energy. According to Sarwani et al. (2019), in
Malaysia, around 4200 tons of methane per year can be produced from
POME. Also, the amount of energy contained in Malaysia-generated
POME would be valued at approximately $ 1 million (Ahmed et al.,
2015). In 2019, 53 POME treatment projects to produce biogas were
registered in the Country. According to Sarwani et al. (2019), 10.7
million m3 of POME were treated and 107,800 tons of methane were
produced.
In parallel, high productions of biogas from wastes were registered in
Thailand, one of the main producers of cassava. Production values of 1.4
million m3 and 62,000 m3 of compressed bio-methane gas were reached,
from cassava and palm industry waste, respectively (Tonrangklang
et al., 2017).
Considering the data of agro-industrial wastes production (Table 1),
the energy content all effluents can be estimated (Equation 1) as approx.
342,758 GWh. This is equivalent to approx. 1.5% of the world energy
consumption in 2019 (23,000 TWh) (ENERDATA, 2020) and could
satisfy the needs of approximately 1.8 million homes with an average
consumption of 180 kWh. Table 4 shows that 62% of the energy is
contained in VW, which is the most abundant effluent, besides having
extremely high organic loads.
E = OL * Cp * V * YE [1]
Where: OL: Organic load (g/L); Cp: Calorific power of methane (CH4)
(802 kJ/mol); V: Effluent volume (L); YE: Methane yield from effluent
(CPW: 0.35 L CH4/g COD (Ubalua, 2007); CW: 0.23 L CH4/g COD
(Prazeres et al., 2012); POME: 0.5 L CH4/g COD (O-Thong et al., 2012);
EMP: 0.19 L CH4/g COD (Kothari et al., 2017); SH: 0.14 L CH4/g COD.
5.2. The potential of agro-industrial wastewaters as raw materials for
other bioproducts
Applying the concept of biorefinery to the processing of agro-
industrial wastewaters is an important approach to increase the rele­
vance of such resources in the local bioeconomy. Biorefineries are fa­
cilities dedicated to the conversion of biomass not only to energy but
also to valuable commercial products. Ideally, the product portfolio is
flexible, and the technologies are robust and easily adaptable to varying
demands. The goals to be pursuit while designing a biorefinery are the
development of cost-beneficial technologies of biomass processing and
conversion, the supply of energy to meet at least the demands of the
plant, the no-generation of waste streams that could cause environ­
mental impacts, the reduction of greenhouse gas emissions and, ulti­
mately, the generation of jobs to promote local socioeconomic
development.
Agro-industrial wastes can be converted to new products mainly
through thermochemical and biological processes. The thermochemical
route includes pyrolysis and gasification, which are carried at extreme
conditions of pressure and temperature and depend on large energy
requirements. Depending on the source of this energy, and the chemical
catalysts used, process sustainability is largely harmed. On the other
hand, biological conversion of agro wastes is carried at milder condi­
tions (even extremophile bioprocesses are carried at much lower tem­
perature than thermochemical processes) and can generate an infinitude
Table 4
Amount of energy contained in the agro-industrial effluents.
Effluents Energy (GWh) % Total energy
CPW 16,645 4.85
VW 211,734 61.77
POME 76,422 22.32
SH 12,202 3.56
CW 24,452 7.13
EMP 1,302 0.4
CPW: cassava processing waste; VW: vinasse wastewater; POME: palm oil mill
effluent; SH: slaughterhouse effluent; CW: cheese whey; EMP: effluent of milk
processing
7
Bioresource Technology 341 (2021) 125795
of products as presented in Fig. 3. The strategy for the valorization of
agro wastes should consider (i) those bio-products with direct applica­
tion within the agro-industry and (ii) those that are produced as a
strategy for market diversification. The agroindustry depends greatly on
the production and supply of raw materials that are cultivated (plant-
based) or grown (animal-based), thus bioproducts that could be directed
to soil fertilization or feed result in direct benefit and can be easily
integrated.
Microbial biomass (originated from bacteria, yeasts, filamentous
fungi and microalgae) is an important source of alternative proteins,
lipids and carbohydrates, and contains bioactive molecules that result in
health improvement of the animals. The fertilizing potential of fer­
mented effluents is related to the content of phytohormones (Shi et al.,
2017), macronutrients for plants (NPK) (Chontal et al., 2019) and
stimulants of the soil microbial community (Hou et al., 2017). The use of
effluent-based bioproducts as fertilizers and feed should guarantee
environmental and animal security in such a way that pre-fermentation
may be considered for effluent valorization, and detoxification in the
case of CPW (presence of cyanogenic compounds), VW (Pb and Zn) and
POME (heavy metals) (Table 2).
Ethanol is a traditional fuel used in the transportation sector, espe­
cially in Brazil, that can be produced through anaerobic fermentation.
However, it is used in flex-fuel cars, not in the heavy vehicles used to
transport raw materials and final production for/to the agroindustry.
Moreover, the economic production of ethanol requires a final concen­
tration of at least 6 ◦ GL, ideally 7–8 ◦ GL (Siqueira et al., 2008) in the
fermentation broth, which results from an initial sugar concentration of
more than 100 g/L, much higher than the typical sugar concentrations of
agro-industrial wastewaters, even after hydrolysis. Butanol is another
liquid fuel that can be produced through the fermentation of agro-
industrial wastes (Behera et al., 2019). Despite not currently used as
transportation fuel, many studies have presented biobutanol as a chal­
lenging and promising fuel for diesel engines (Tucki et al., 2020). This
biofuel has higher energy content, lower volatility and can be mixed
with gasoline or diesel in higher proportions as compared to ethanol.
There are still many challenges related to the biochemical production
and purification of butanol, and final concentrations in the fermentation
broth are typically low, in the order of 5–15 g/L (Kushwaha et al., 2019).
But considering that the substrate costs may represent even 60% of the
total production costs, and that the necessary initial sugar concentra­
tions are not considerably high, the use of agro-industrial wastewaters
may be interesting in this case. Even the direct production of butanol
from cellulose has been reported, using engineered Clostridium cellulo­
lyticum (Gaida et al., 2016).
Other value-added products (Fig. 3), such as enzymes (Ohimain
et al., 2013), protein (Nitayavardhana et al., 2013), organic acids (do
Nascimento Junior et al., 2021), volatile fatty acids (Damasceno et al.,
2003), lipids (Fernandes et al., 2017), biosurfactants (Barros et al., 2008;
Nitschke and Pastore, 2003, 2006), biolubricants (Boviatsi et al., 2020),
bioflocculants (Bukhari et al., 2017), acetone (Fernandes et al., 2017),
carotenoids (Santana et al., 2017), exopolysaccharides (Ventorino et al.,
2019), polyesters (Koller et al., 2008), penicillin (Wu et al., 2009), plant
hormones (Chatterjee et al., 2008), vitamins (Hugenschmidt et al.,
2010) and volatile flavors (Marques et al., 2021) have already been
produced from agro-industrial liquid wastes at laboratory scale. Here,
the main challenges are, besides achieving high yields and productivity,
developing efficient downstream processing techniques that allow the
economic scale-up of such processes, considering that these residues
have complex compositions and are usually very diluted.
6. Research needs and future directions
Agro-industrial effluents constitute a raw material that is still under-
utilized and that, through appropriate technologies, could contribute
greatly to decarbonize the planet, whether through the production of
biofuels or biochemical products. The energy contained in the agro-
W.J. Martinez-Burgos et al.
Fig. 3. Different bio-products obtained from agro-industrial effluents. CPW: cassava processing waste; SH: slaughterhouse effluent; POME: palm oil mill
effluent; VW: vinasse wastewater; EMP: effluent of milk processing.
industrial effluents can supply about 1.8 million homes. The bio­
processing of agro-industrial wastewaters results in the optimization of
the natural, human, and economic resources applied in food and feed
production. Moreover, it reduces environmental impacts and promotes
the circular bioeconomy.
Despite considered a path of no return in the modern economy,
working with agro-industrial wastewaters is challenging especially due
to the variable composition that depends mainly on the raw material’s
composition (which varies according to soil type, culture management,
climate, etc.) and industrial processing. This results in considerable
variation on productivity and deserves attention in technology devel­
opment. The lack of standardization in the units of analysis many times
hinders the comparison between studies and technologies (such as
molH2/L, mLH2/L and mLH2/gCOD, in the case of biohydrogen).
At the same time that the high production rate of agro-industrial
wastes is advantageous in terms of raw material availability, it im­
poses challenges when medium supplementation, sterilization and pre-
treatments are mandatory. In these cases, both economic viability and
sustainability can be threatened, and economic evaluations and life
cycle assessments are crucial. The production of energy and fertilizers
are the simplest strategies to integrate the use of agro-industrial wastes
in the agroindustry. Anaerobic digestion of liquid and thermal pro­
cessing of solid wastes are the most mature technologies at commercial
scale, but the demand of sustainable production in all areas will continue
to drive the scientific development towards the production of several
biomolecules.
7. Conclusion
Agro-industrial wastewaters are produced in large volumes and
present high concentrations of organic matter, nitrogen, phosphorus,
and potassium. These effluents can cause great impacts in different
ecological systems. The most rational (and economical) way for their
disposal is by associating them with the production of energy and high
commercial value bioproducts by applying biorefinery concepts. Biogas
production, especially due to its versatility and ease of integration into
various industrial systems, is the most explored pathway, but the
8
Bioresource Technology 341 (2021) 125795
obtaining of other value-added products can increase the economic
profitability of such processes and create opportunities for local
development.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgements
This work was supported by the Brazilian National Council for Sci­
entific and Technological Development (CNPq) and the Coordination of
Improvement of Higher Education Personnel (CAPES).
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